Editor’s Choice Articles

Editor’s Choice articles are based on recommendations by the scientific editors of MDPI journals from around the world. Editors select a small number of articles recently published in the journal that they believe will be particularly interesting to readers, or important in the respective research area. The aim is to provide a snapshot of some of the most exciting work published in the various research areas of the journal.

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29 pages, 8564 KB  
Review
Comprehensive Review on DNA Hydrogels and DNA Origami-Enabled Wearable and Implantable Biosensors
by Man Li and Joonho Bae
Biosensors 2025, 15(12), 819; https://doi.org/10.3390/bios15120819 - 18 Dec 2025
Cited by 9 | Viewed by 2270
Abstract
DNA nanoparticles have emerged as potent platforms for wearable and implantable biosensors owing to their molecular programmability, biocompatibility, and structural precision. This study delineates two principal categories of DNA-based sensing materials, DNA hydrogels and DNA origami, and encapsulates their fabrication methodologies, sensing mechanisms, [...] Read more.
DNA nanoparticles have emerged as potent platforms for wearable and implantable biosensors owing to their molecular programmability, biocompatibility, and structural precision. This study delineates two principal categories of DNA-based sensing materials, DNA hydrogels and DNA origami, and encapsulates their fabrication methodologies, sensing mechanisms, and applications at the device level. DNA hydrogels serve as pliable, aqueous signal transduction mediums exhibiting stimulus-responsive characteristics, facilitating applications such as sweat-based cytokine detection with limits of detection as low as pg·mL−1 and microneedle-integrated hydrogels for femtomolar miRNA sensing. DNA origami offers nanometer-scale spatial precision that improves electrochemical, optical, and plasmonic biosensing, as shown by origami-facilitated luminous nucleic acid detection and ultrasensitive circulating tumor DNA assays with fM-level sensitivity. Emerging integration technologies, such as flexible electronics, microfluidics, and wireless readout, are examined, alongside prospective developments in AI-assisted DNA design and materials produced from synthetic biology. This study offers a thorough and practical viewpoint on the progression of DNA nanotechnology for next-generation wearable and implantable biosensing devices. Full article
(This article belongs to the Section Wearable Biosensors)
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36 pages, 6926 KB  
Review
AI-Integrated Micro/Nanorobots for Biomedical Applications: Recent Advances in Design, Fabrication, and Functions
by Prashant Kishor Sharma and Chia-Yuan Chen
Biosensors 2025, 15(12), 793; https://doi.org/10.3390/bios15120793 - 2 Dec 2025
Cited by 18 | Viewed by 4422
Abstract
The integration of artificial intelligence (AI) and micro/nanorobotics is fundamentally reshaping biosensing by enabling autonomous, adaptive, and high-resolution biological analysis. These miniaturized robotic systems fabricated using advanced techniques such as photolithography, soft lithography, nanoimprinting, 3D printing, and self-assembly can navigate complex biological environments [...] Read more.
The integration of artificial intelligence (AI) and micro/nanorobotics is fundamentally reshaping biosensing by enabling autonomous, adaptive, and high-resolution biological analysis. These miniaturized robotic systems fabricated using advanced techniques such as photolithography, soft lithography, nanoimprinting, 3D printing, and self-assembly can navigate complex biological environments to perform targeted sensing, diagnostics, and therapeutic delivery. AI-driven algorithms, mainly those in machine learning (ML) and deep learning (DL), act as the brains of the operation, allowing for sophisticated modeling, genuine real-time control, and complex signal interpretation. This review focuses recent advances in the design, fabrication, and functional integration of AI-enabled micro/nanorobots for biomedical sensing. Applications that demonstrate their potential range from quick point-of-care diagnostics and in vivo biosensing to next-generation organ-on-chip systems and truly personalized medicine. We also discuss key challenges in scalability, energy autonomy, data standardization, and closed-loop control. Collectively, these advancements are paving the way for intelligent, responsive, and clinically transformative biosensing systems. Full article
(This article belongs to the Section Biosensors and Healthcare)
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14 pages, 2020 KB  
Review
Wearable Sensors for Precise Exercise Monitoring and Analysis
by Bo Su, Fengyu Li and Bingtian Su
Biosensors 2025, 15(11), 734; https://doi.org/10.3390/bios15110734 - 3 Nov 2025
Cited by 12 | Viewed by 5565
Abstract
The adoption of wearable sensors for precision training has accelerated in recent years, yet most studies and reviews remain device- or feasibility-centric and lack a field-ready decision framework. This review organizes wearable sensing across four monitoring dimensions—physiological, kinematic, biochemical, and dynamic—and maps them [...] Read more.
The adoption of wearable sensors for precision training has accelerated in recent years, yet most studies and reviews remain device- or feasibility-centric and lack a field-ready decision framework. This review organizes wearable sensing across four monitoring dimensions—physiological, kinematic, biochemical, and dynamic—and maps them onto three training pillars: physical, technical, and tactical. From the perspectives of athletes and coaches, we operationalize quality control, threshold, and feedback loop to translate measurement into action. We critically appraise key limitations, including signal robustness under high-intensity motion, inter-individual variability and limited model generalizability, cross-device data fusion and latency, battery life and wearability, privacy and data ownership, and limited accessibility beyond elite settings. Looking ahead, we advocate a shift from mere multidimensional measurement to a verifiable, reusable, and deployable precision-training ecosystem that delivers actionable metrics and clear decision support for practitioners. Full article
(This article belongs to the Special Issue Wearable Biosensors and Health Monitoring)
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56 pages, 7355 KB  
Review
Carbon Nanomaterial-Based Electrochemical Biosensors for Alzheimer’s Disease Biomarkers: Progress, Challenges, and Future Perspectives
by Berfin Şak, Helena B. A. Sousa and João A. V. Prior
Biosensors 2025, 15(10), 684; https://doi.org/10.3390/bios15100684 - 9 Oct 2025
Cited by 16 | Viewed by 4448
Abstract
Alzheimer’s disease (AD) requires early and accurate identification of affected brain regions, which can be achieved through the detection of specific biomarkers to enable timely intervention. Carbon nanomaterials (CNMs), including graphene derivatives, carbon nanotubes, graphitic carbon nitride, carbon black, fullerenes, and carbon dots, [...] Read more.
Alzheimer’s disease (AD) requires early and accurate identification of affected brain regions, which can be achieved through the detection of specific biomarkers to enable timely intervention. Carbon nanomaterials (CNMs), including graphene derivatives, carbon nanotubes, graphitic carbon nitride, carbon black, fullerenes, and carbon dots, offer high conductivity, large electroactive surface area, and versatile surface chemistry that enhance biosensor performance. While such properties benefit a wide range of transduction principles (e.g., electrochemical, optical, and plasmonic), this review focuses on their role in electrochemical biosensors. This review summarizes CNM-based electrochemical platforms reported from 2020 to mid-2025, employing aptamers, antibodies, and molecularly imprinted polymers for AD biomarker detection. Covered topics include fabrication strategies, transduction formats, analytical performance in complex matrices, and validation. Reported devices achieve limits of detection from the femtomolar to picogram per milliliter range, with linear ranges typically spanning 2–3 orders of magnitude (e.g., from femtomolar to picomolar, or from picogram to nanogram per milliliter levels). They exhibit high selectivity against common interferents such as BSA, glucose, uric acid, ascorbic acid, dopamine, and non-target peptides, along with growing capabilities for multiplexing and portable operation. Remaining challenges include complex fabrication, limited long-term stability and reproducibility data, scarce clinical cohort testing, and sustainability issues. Opportunities for scalable production and integration into point-of-care workflows are outlined. Full article
(This article belongs to the Special Issue Nano/Micro Biosensors for Biomedical Applications (2nd Edition))
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33 pages, 781 KB  
Review
Recent Advances in Electrochemical Sensors for the Detection of Anti-Inflammatory and Antibiotic Drugs: A Comprehensive Review
by Gisele Afonso Bento Mello, Stephen Rathinaraj Benjamin, Fábio de Lima and Rosa F. Dutra
Biosensors 2025, 15(10), 676; https://doi.org/10.3390/bios15100676 - 8 Oct 2025
Cited by 29 | Viewed by 5733
Abstract
Electrochemical sensors have emerged as powerful analytical tools for the detection of anti-inflammatory and antibiotic drugs due to their high sensitivity, rapid response, and cost-effectiveness compared to conventional chromatographic and spectrophotometric methods. This review highlights recent advances in electrode materials, surface modification strategies, [...] Read more.
Electrochemical sensors have emerged as powerful analytical tools for the detection of anti-inflammatory and antibiotic drugs due to their high sensitivity, rapid response, and cost-effectiveness compared to conventional chromatographic and spectrophotometric methods. This review highlights recent advances in electrode materials, surface modification strategies, and signal amplification approaches for quantifying nonsteroidal anti-inflammatory drugs (NSAIDs) and various antibiotic classes, including sulfonamides, tetracyclines, macrolides, and quinolones. Particular attention is given to nanostructured carbon-based materials, metal nanoparticles, and polymer composites that enhance electron transfer, improve selectivity, and lower limits of detection (LODs). The analytical performance of different electrochemical techniques such as cyclic voltammetry, differential pulse voltammetry, and square-wave voltammetry is critically compared across various drug targets. Trends indicate that hybrid nanomaterial-modified electrodes consistently achieve sub-micromolar detection limits in biological and environmental samples, offering potential for point-of-care diagnostics and environmental monitoring. Current challenges include improving sensor stability, mitigating fouling effects, and ensuring reproducibility in complex matrices. Future research should focus on integrated, miniaturized sensing platforms capable of multiplex detection, paving the way for rapid, portable, and sustainable analytical solutions in pharmaceutical and biomedical applications. Full article
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29 pages, 2876 KB  
Review
Exhaled Aldehydes and Ketones as Biomarkers of Lung Cancer and Diabetes: Review of Sensor Technologies for Early Disease Diagnosis
by Rafał Kiejzik, Tomasz Wasilewski and Wojciech Kamysz
Biosensors 2025, 15(10), 668; https://doi.org/10.3390/bios15100668 - 3 Oct 2025
Cited by 9 | Viewed by 2865
Abstract
Exhaled breath (EB) contains numerous volatile organic compounds (VOCs) that can reflect pathological metabolic processes, making breath analysis a promising non-invasive diagnostic approach. In particular, volatile aldehydes and ketones have been identified as disease biomarkers in EB. Gas sensors are expected to play [...] Read more.
Exhaled breath (EB) contains numerous volatile organic compounds (VOCs) that can reflect pathological metabolic processes, making breath analysis a promising non-invasive diagnostic approach. In particular, volatile aldehydes and ketones have been identified as disease biomarkers in EB. Gas sensors are expected to play a crucial role in the diagnosis of numerous diseases at an early stage. Among the various available approaches, sensors stand out as especially attractive tools for diagnosing diseases such as lung cancer (LC) and diabetes, due to their affordability and operational simplicity. There is an urgent need in the field of disease detection for the development of affordable, non-invasive, and user-friendly sensors capable of detecting various biomarkers. Devices of the new generation should also demonstrate high repeatability of measurements and extended operational stability of the employed sensors. Due to these demands, the past few years have seen significant advancements in the development and implementation of electronic noses (ENs), which are composed of an array of sensors for the determination of VOCs present in EB. To meet these requirements, the development and integration of advanced receptor coatings on sensor transducers is essential. These coatings include nanostructured materials, molecularly imprinted polymers, and bioreceptors, which collectively enhance selectivity, sensitivity, and operational stability. However, reliable biomarker detection in point-of-care (PoC) mode remains a significant challenge, constrained by several factors. This review provides a comprehensive and critical evaluation of recent studies demonstrating that the detection of VOCs using gas sensor platforms enables disease detection and can be implemented in PoC mode. Full article
(This article belongs to the Special Issue Functional Materials for Biosensing Applications)
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38 pages, 2063 KB  
Review
Nanostructured Materials in Glucose Biosensing: From Fundamentals to Smart Healthcare Applications
by Rajaram Rajamohan and Seho Sun
Biosensors 2025, 15(10), 658; https://doi.org/10.3390/bios15100658 - 2 Oct 2025
Cited by 11 | Viewed by 3723
Abstract
The rapid development of nanotechnology has significantly transformed the design and performance of glucose biosensors, leading to enhanced sensitivity, selectivity, and real-time monitoring capabilities. This review highlights recent advances in glucose-sensing platforms facilitated by nanomaterials, including metal and metal oxide nanoparticles, carbon-based nanostructures, [...] Read more.
The rapid development of nanotechnology has significantly transformed the design and performance of glucose biosensors, leading to enhanced sensitivity, selectivity, and real-time monitoring capabilities. This review highlights recent advances in glucose-sensing platforms facilitated by nanomaterials, including metal and metal oxide nanoparticles, carbon-based nanostructures, two-dimensional materials, and metal–organic frameworks (MOFs). The integration of these nanoscale materials into electrochemical, optical, and wearable biosensors has addressed longstanding challenges associated with enzyme stability, detection limits, and invasiveness. Special emphasis is placed on non-enzymatic glucose sensors, flexible and wearable devices, and hybrid nanocomposite systems. The multifunctional properties of nanomaterials, such as large surface area, excellent conductivity, and biocompatibility, have enabled the development of next-generation sensors for clinical, point-of-care, and personal healthcare applications. The review also discusses emerging trends such as biodegradable nanosensors, AI-integrated platforms, and smart textiles, which are poised to drive the future of glucose monitoring toward more sustainable and personalized healthcare solutions. Full article
(This article belongs to the Special Issue Recent Advances in Glucose Biosensors)
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25 pages, 6394 KB  
Review
Artificial Intelligence-Assisted Nanosensors for Clinical Diagnostics: Current Advances and Future Prospects
by Shuo Yin
Biosensors 2025, 15(10), 656; https://doi.org/10.3390/bios15100656 - 1 Oct 2025
Cited by 23 | Viewed by 4986
Abstract
The integration of artificial intelligence (AI) with various diagnostic nanosensors has opened up new horizons in clinics recently. AI technology offers enhanced sensitivity, accuracy, specificity, and real-time analysis for disease diagnostics. This review focuses on the recent advances in AI-assisted nanosensors for the [...] Read more.
The integration of artificial intelligence (AI) with various diagnostic nanosensors has opened up new horizons in clinics recently. AI technology offers enhanced sensitivity, accuracy, specificity, and real-time analysis for disease diagnostics. This review focuses on the recent advances in AI-assisted nanosensors for the diagnosis of different diseases in clinical applications. Critical roles of AI in sensor design, optimization, signal processing, and clinical decision support are highlighted. Furthermore, challenges such as limited datasets, regulatory hurdles, and data privacy are discussed, along with future opportunities. This review aims to provide a comprehensive introduction and perspectives on how AI-driven nanosensors are transforming clinical diagnostics and shaping the future of precise medicine. Full article
(This article belongs to the Special Issue Nanosensors for Bioanalysis)
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54 pages, 3531 KB  
Review
Designing the Future of Biosensing: Advances in Aptamer Discovery, Computational Modeling, and Diagnostic Applications
by Robert G. Jesky, Louisa H. Y. Lo, Ryan H. P. Siu and Julian A. Tanner
Biosensors 2025, 15(10), 637; https://doi.org/10.3390/bios15100637 - 24 Sep 2025
Cited by 24 | Viewed by 7327
Abstract
Recent advances in computational tools, particularly machine learning (ML), deep learning (DL), and structure-based modeling, are transforming aptamer research by accelerating discovery and enhancing biosensor development. This review synthesizes progress in predictive algorithms that model aptamer–target interactions, guide in silico sequence optimization, and [...] Read more.
Recent advances in computational tools, particularly machine learning (ML), deep learning (DL), and structure-based modeling, are transforming aptamer research by accelerating discovery and enhancing biosensor development. This review synthesizes progress in predictive algorithms that model aptamer–target interactions, guide in silico sequence optimization, and streamline design workflows for both laboratory and point-of-care diagnostic platforms. We examine how these approaches improve key aspects of aptasensor development, such as aptamer selection, sensing surface immobilization, signal transduction, and molecular architecture, which contribute to greater sensitivity, specificity, and real-time diagnostic capabilities. Particular attention is given to illuminating the technological and experimental advances in structure-switching aptamers, dual-aptamer systems, and applications in electrochemical, optical, and lateral flow platforms. We also discuss current challenges such as the standardization of datasets and interpretability of ML models and highlight future directions that will support the translation of aptamer-based biosensors into scalable, point-of-care and clinically deployable diagnostic solutions. Full article
(This article belongs to the Special Issue Nucleic Acid Aptamer-Based Bioassays)
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25 pages, 13991 KB  
Review
Progress and Prospects in FRET for the Investigation of Protein–Protein Interactions
by Yue Zhang, Xinyue Ma, Meihua Zhu, Vivien Ya-Fan Wang and Jiajia Guo
Biosensors 2025, 15(9), 624; https://doi.org/10.3390/bios15090624 - 19 Sep 2025
Cited by 8 | Viewed by 4950
Abstract
Protein–protein interactions (PPIs) play a crucial role in various biological processes, including signal transduction, transcriptional regulation, and metabolic pathways. Over the years, many methods have been developed to study PPIs, such as yeast two-hybrid (Y2H), co-immunoprecipitation (Co-IP), pull-down assays, and surface plasmon resonance [...] Read more.
Protein–protein interactions (PPIs) play a crucial role in various biological processes, including signal transduction, transcriptional regulation, and metabolic pathways. Over the years, many methods have been developed to study PPIs, such as yeast two-hybrid (Y2H), co-immunoprecipitation (Co-IP), pull-down assays, and surface plasmon resonance (SPR). However, each of these techniques has its own limitations, including false positives, a lack of specific binding partners, and restricted interaction zones. Fluorescence resonance energy transfer (FRET) has emerged as a powerful technique for investigating PPIs, offering several advantages over traditional methods. Recent advancements in fluorescence microscopy have further enhanced its application in PPI studies. In this review, we summarize recent developments in FRET-based approaches and their applications in PPIs research over the past five years, including conventional FRET, time-resolved FRET (TR-FRET), fluorescence lifetime imaging microscopy-FRET (FLIM-FRET), single-molecule FRET (smFRET), fluorescence cross-correlation spectroscopy FRET (FCCS-FRET), and provide guidance on selecting the most appropriate method for PPIs studies. Full article
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29 pages, 4099 KB  
Review
Deep Learning-Driven Multimodal Integration of miRNA and Radiomic for Lung Cancer Diagnosis
by Yuanyuan Chen, Dikang Chen, Xiaohui Liu, Hui Jiang and Xuemei Wang
Biosensors 2025, 15(9), 610; https://doi.org/10.3390/bios15090610 - 16 Sep 2025
Cited by 10 | Viewed by 3239
Abstract
Lung cancer remains one of the most common and deadly malignancies worldwide. Current diagnosis and staging primarily rely on biopsy techniques, which fail to comprehensively characterize the molecular profiles and tumor microenvironment. Current studies demonstrate the promising performance (AUC = 82%) of miRNA-based [...] Read more.
Lung cancer remains one of the most common and deadly malignancies worldwide. Current diagnosis and staging primarily rely on biopsy techniques, which fail to comprehensively characterize the molecular profiles and tumor microenvironment. Current studies demonstrate the promising performance (AUC = 82%) of miRNA-based predictive models, but exclusive reliance on miRNA signatures is limited by incomplete capture of tumor heterogeneity. Integrating imaging and genomic data can further enhance model accuracy, with functional nanomaterials serving as core advanced biosensing platforms to bridge miRNA sensing and radiomic fusion. Consequently, integrating imaging and genomic data can further enhance model accuracy. Recent research employing DenseNet architecture for the multimodal fusion of miRNA and radiomic features achieved an AUC of 0.98 with 85.7% sensitivity. This review summarizes advances in miRNA biomarkers, deep learning-driven radiogenomics, and critical roles of functional nanomaterials in biosensing-enabled multimodal integration, along with challenges and future directions for clinical translation. Full article
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37 pages, 2546 KB  
Review
POC Sensor Systems and Artificial Intelligence—Where We Are Now and Where We Are Going?
by Prashanthi Kovur, Krishna M. Kovur, Dorsa Yahya Rayat and David S. Wishart
Biosensors 2025, 15(9), 589; https://doi.org/10.3390/bios15090589 - 8 Sep 2025
Cited by 16 | Viewed by 4725
Abstract
Integration of machine learning (ML) and artificial intelligence (AI) into point-of-care (POC) sensor systems represents a transformative advancement in healthcare. This integration enables sophisticated data analysis and real-time decision-making in emergency and intensive care settings. AI and ML algorithms can process complex biomedical [...] Read more.
Integration of machine learning (ML) and artificial intelligence (AI) into point-of-care (POC) sensor systems represents a transformative advancement in healthcare. This integration enables sophisticated data analysis and real-time decision-making in emergency and intensive care settings. AI and ML algorithms can process complex biomedical data, improve diagnostic accuracy, and enable early disease detection for better patient outcomes. Predictive analytics in POC devices supports proactive healthcare by analyzing data to forecast health issues and facilitating early intervention and personalized treatment. This review covers the key areas of ML and AI integration in POC devices, including data analysis, pattern recognition, real-time decision support, predictive analytics, personalization, automation, and workflow optimization. Examples of current POC devices that use ML and AI include AI-powered blood glucose monitors, portable imaging devices, wearable cardiac monitors, AI-enhanced infectious disease detection, and smart wound care sensors are also discussed. The review further explores new directions for POC sensors and ML integration, including mental health monitoring, nutritional monitoring, metabolic health tracking, and decentralized clinical trials (DCTs). We also examined the impact of integrating ML and AI into POC devices on healthcare accessibility, efficiency, and patient outcomes. Full article
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40 pages, 3625 KB  
Review
Graphene-Based Biosensors: Enabling the Next Generation of Diagnostic Technologies—A Review
by John Paolo Ramoso, Manoochehr Rasekh and Wamadeva Balachandran
Biosensors 2025, 15(9), 586; https://doi.org/10.3390/bios15090586 - 6 Sep 2025
Cited by 35 | Viewed by 11508
Abstract
Graphene, a two-dimensional carbon material with a hexagonal lattice structure, possesses remarkable properties. Exceptional electrical conductivity, mechanical strength, and high surface area that make it a powerful platform for biosensing applications. Its sp2-hybridised network facilitates efficient electron mobility and enables diverse [...] Read more.
Graphene, a two-dimensional carbon material with a hexagonal lattice structure, possesses remarkable properties. Exceptional electrical conductivity, mechanical strength, and high surface area that make it a powerful platform for biosensing applications. Its sp2-hybridised network facilitates efficient electron mobility and enables diverse surface functionalisation through bio-interfacing. This review highlights the core detection mechanisms in graphene-based biosensors. Optical sensing techniques, such as surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS), benefit significantly from graphene’s strong light–matter interaction, which enhances signal sensitivity. Although graphene itself lacks intrinsic piezoelectricity, its integration with piezoelectric substrates can augment the performance of piezoelectric biosensors. In electrochemical sensing, graphene-based electrodes support rapid electron transfer, enabling fast response times across a range of techniques, including impedance spectroscopy, amperometry, and voltammetry. Graphene field-effect transistors (GFETs), which leverage graphene’s high carrier mobility, offer real-time, label-free, and highly sensitive detection of biomolecules. In addition, the review also explores multiplexed detection strategies vital for point-of-care diagnostics. Graphene’s nanoscale dimensions and tunable surface chemistry facilitate both array-based configurations and the simultaneous detection of multiple biomarkers. This adaptability makes graphene an ideal material for compact, scalable, and accurate biosensor platforms. Continued advancements in graphene biofunctionalisation, sensing modalities, and integrated multiplexing are driving the development of next-generation biosensors with superior sensitivity, selectivity, and diagnostic reliability. Full article
(This article belongs to the Special Issue Novel Graphene-Based Biosensors for Biomedical Applications)
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22 pages, 3921 KB  
Article
A Sensitive Electrochemical Cholinesterase-Inhibiting Biosensor for Organophosphorus Pesticides Based on Ti3C2TX MXene Quantum Dots
by Nisha Makani, Jett Wu, Jose Florentino, Cecilia F. Chafin, Bhoj Gautam, Shirley Chao and Shubo Han
Biosensors 2025, 15(9), 575; https://doi.org/10.3390/bios15090575 - 2 Sep 2025
Cited by 18 | Viewed by 2555
Abstract
Organophosphorus pesticides (OPs) pose significant environmental and health risks due to their widespread use and toxicity, primarily by inhibiting acetylcholinesterase. Traditional detection methods are often slow and costly, highlighting the urgent need for advanced, sensitive, and accessible technologies. This study developed a highly [...] Read more.
Organophosphorus pesticides (OPs) pose significant environmental and health risks due to their widespread use and toxicity, primarily by inhibiting acetylcholinesterase. Traditional detection methods are often slow and costly, highlighting the urgent need for advanced, sensitive, and accessible technologies. This study developed a highly sensitive electrochemical cholinesterase-inhibiting biosensor for OP pesticides, utilizing Ti3C2Tx MXene Quantum Dots (MQDs), which was synthesized via a hydrothermal method. The biosensor’s performance was characterized using electrochemical impedance spectroscopy, differential pulse voltammetry (DPV), and cyclic voltammetry. DPV proved to be the optimal technique, exhibiting an ultralow detection limit of 1 × 10−17 M and a wide linear range (10−14–10−8 M) for chlorpyrifos (a model OP) with an estimated inhibition constant of 62 nM. The biosensor demonstrated high selectivity for OPs (chlorpyrifos, acephate, glyphosate) over a non-target pyrethroid (permethrin), confirmed by distinct electrochemical signatures and compared to in vitro cholinergic activity assays in bean beetle homogenates. The enhanced performance is attributed to the high surface-to-volume ratio, quantum confinement effects, and superior conductivity of the MQDs, as well as the robust enzyme immobilization facilitated by glutaraldehyde cross-linking and a chitosan matrix. This work presents a promising platform for rapid, sensitive, and selective detection of OP pesticides, with potential applications in environmental monitoring and public health protection. Full article
(This article belongs to the Section Biosensor Materials)
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44 pages, 10149 KB  
Review
A Review of Machine Learning-Assisted Gas Sensor Arrays in Medical Diagnosis
by Yueting Yu, Xin Cao, Chenxi Li, Mingyue Zhou, Tianyu Liu, Jiang Liu and Lu Zhang
Biosensors 2025, 15(8), 548; https://doi.org/10.3390/bios15080548 - 20 Aug 2025
Cited by 29 | Viewed by 7586
Abstract
Volatile organic compounds (VOCs) present in human exhaled breath have emerged as promising biomarkers for non-invasive disease diagnosis. However, traditional VOC detection technology that relies on large instruments is not widely used due to high costs and cumbersome testing processes. Machine learning-assisted gas [...] Read more.
Volatile organic compounds (VOCs) present in human exhaled breath have emerged as promising biomarkers for non-invasive disease diagnosis. However, traditional VOC detection technology that relies on large instruments is not widely used due to high costs and cumbersome testing processes. Machine learning-assisted gas sensor arrays offer a compelling alternative by enabling the accurate identification of complex VOC mixtures through collaborative multi-sensor detection and advanced algorithmic analysis. This work systematically reviews the advanced applications of machine learning-assisted gas sensor arrays in medical diagnosis. The types and principles of sensors commonly employed for disease diagnosis are summarized, such as electrochemical, optical, and semiconductor sensors. Machine learning methods that can be used to improve the recognition ability of sensor arrays are systematically listed, including support vector machines (SVM), random forests (RF), artificial neural networks (ANN), and principal component analysis (PCA). In addition, the research progress of sensor arrays combined with specific algorithms in the diagnosis of respiratory, metabolism and nutrition, hepatobiliary, gastrointestinal, and nervous system diseases is also discussed. Finally, we highlight current challenges associated with machine learning-assisted gas sensors and propose feasible directions for future improvement. Full article
(This article belongs to the Special Issue AI-Enabled Biosensor Technologies for Boosting Medical Applications)
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24 pages, 3609 KB  
Review
Droplet-Based Microfluidics in Single-Bacterium Analysis: Advancements in Cultivation, Detection, and Application
by Haiyan Ma, Yuewen Zhang, Ren Shen and Yanwei Jia
Biosensors 2025, 15(8), 535; https://doi.org/10.3390/bios15080535 - 15 Aug 2025
Cited by 12 | Viewed by 5732
Abstract
Microorganisms exhibit remarkable diversity, making their comprehensive characterization essential for understanding ecosystem functioning and safeguarding human health. However, traditional culture-based methods entail inherent limitations for resolving microbial heterogeneity, isolating slow-growing microorganisms, and accessing uncultivated microbes. Conversely, droplet-based microfluidics enables a high-throughput and precise [...] Read more.
Microorganisms exhibit remarkable diversity, making their comprehensive characterization essential for understanding ecosystem functioning and safeguarding human health. However, traditional culture-based methods entail inherent limitations for resolving microbial heterogeneity, isolating slow-growing microorganisms, and accessing uncultivated microbes. Conversely, droplet-based microfluidics enables a high-throughput and precise platform for single-bacterium manipulation by physically isolating individual cells within microdroplets. This technology presents a transformative approach to overcoming the constraints of conventional techniques. This review outlines the fundamental principles, recent research advances, and key application domains of droplet-based microfluidics, with a particular focus on innovations in single-bacterium encapsulation, sorting, cultivation, and functional analysis. Applications such as antibiotic susceptibility testing, enzyme-directed evolution screening, microbial interaction studies, and the cultivation of novel bacterial species are discussed, underscoring the technology’s broad potential in microbiological research and biotechnology. Full article
(This article belongs to the Special Issue Biosensors Based on Microfluidic Devices—2nd Edition)
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16 pages, 6084 KB  
Article
First Design of a Contact Lens for Diagnosis of Dehydration
by Kundan Sivashanmugan, Reece E. Albert and Joseph R. Lakowicz
Biosensors 2025, 15(8), 532; https://doi.org/10.3390/bios15080532 - 14 Aug 2025
Cited by 2 | Viewed by 1379
Abstract
Dehydration is a serious medical problem for elderly patients and young children. The most widely used diagnostics are measurements of sodium ion (Na+) and potassium ion (K+) in blood serum. Dehydration is difficult to diagnose even by trained health [...] Read more.
Dehydration is a serious medical problem for elderly patients and young children. The most widely used diagnostics are measurements of sodium ion (Na+) and potassium ion (K+) in blood serum. Dehydration is difficult to diagnose even by trained health care professionals because the body compensates to maintain the appearance of skin. These measurements required a blood draw because specific tests are generally not available for only Na+ and K+. The blood samples are analyzed by an electrolyte panel (EP) or a basic metabolic panel (BMP). Most hospitals limit EP and BMP to one per day to control costs. More frequent measurements of Na+ and K+ are needed, especially during rehydration. We designed a dehydration contact lens that can provide the Na+ and K+ concentrations as needed or for continuous monitoring. The measurements are obtained from the fluorescent lifetime or wavelength-ratiometric intensities of the Na+- and K+-sensitive fluorophores. The dehydration contact lens does not contain electronic components and are inexpensive to prepare. Full article
(This article belongs to the Special Issue Advanced Fluorescence Biosensors)
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14 pages, 5194 KB  
Article
Drying-Induced Salt Deposition Patterns as a Tool for Label-Free Protein Quantification
by Arturo Patrone-Garcia, Miquel Avella-Oliver and Ángel Maquieira
Biosensors 2025, 15(8), 520; https://doi.org/10.3390/bios15080520 - 9 Aug 2025
Cited by 2 | Viewed by 1409
Abstract
This work reports a label-free analytical strategy based on protein-induced modulation of salt crystallization patterns upon drying. This method relies on the consistent observation that protein-containing saline samples produce distinct salt deposition morphologies compared to protein-free controls. The work first demonstrates the concept [...] Read more.
This work reports a label-free analytical strategy based on protein-induced modulation of salt crystallization patterns upon drying. This method relies on the consistent observation that protein-containing saline samples produce distinct salt deposition morphologies compared to protein-free controls. The work first demonstrates the concept of this phenomenon and characterizes the structural features of the resulting salt patterns. Then, systematic experiments with different solution compositions, substrates, surface coatings, and protein types confirm the generality of this differential deposition behavior and its dependence on total protein concentration. Two complementary measurement approaches are evaluated: a custom laser-scattering setup for optical attenuation measurements and a digital image analysis method based on pixel intensity distributions. Both strategies enable quantitative protein detection in simple (casein) and complex (human serum) samples, offering good correlations between signal and concentration and detection limits in the range of 2–18 µg·mL−1 for digital image analysis and 162–205 µg·mL−1 for optical attenuation measurements. These findings introduce an appealing paradigm for protein quantification exploiting drying-mediated crystallization phenomena, with potential for simple and label-free bioanalytical assays. Full article
(This article belongs to the Special Issue Optical Sensors for Biological Detection)
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40 pages, 8156 KB  
Review
Advances in the Direct Nanoscale Integration of Molecularly Imprinted Polymers (MIPs) with Transducers for the Development of High-Performance Nanosensors
by Ibrar Muhammad Asif, Tiziano Di Giulio, Francesco Gagliani, Cosimino Malitesta and Elisabetta Mazzotta
Biosensors 2025, 15(8), 509; https://doi.org/10.3390/bios15080509 - 6 Aug 2025
Cited by 13 | Viewed by 4340
Abstract
Molecularly imprinted polymers (MIPs) have emerged as robust, cost-effective analogues of bioreceptors, offering high selectivity and stability. When applied in sensors, one key step is the integration of MIPs with the transducer, which critically affects sensor performance. Demanding challenges come when such integration [...] Read more.
Molecularly imprinted polymers (MIPs) have emerged as robust, cost-effective analogues of bioreceptors, offering high selectivity and stability. When applied in sensors, one key step is the integration of MIPs with the transducer, which critically affects sensor performance. Demanding challenges come when such integration involves nanoscaling processes, meaning that the transducer is nanostructured or the MIP itself is nanosized on a bulk transducer. In both cases, the integration results in the development of nanosensors, with advantages arising from the nanoscale, such as a high MIP surface-to-volume ratio, with surface-located, easily accessible binding sites, fast binding kinetics, and, thus, a rapid sensor response. Major advantages come also from nanostructured transducers, with nanoscale geometry enabling highly sensitive signal generation processes, not allowed on their bulk counterparts. In this review, we discuss advances in imprinting technologies, focusing on techniques that, enabling the nanoscale control of MIP synthesis, are conveniently applied to directly integrate MIPs with nanosensors in a one-step process. Two main approaches are reviewed, consisting in MIP nanostructuring on bulk transducers and in the direct growth of MIPs on nanotransducers, highlighting how different strategies achieve good conformity at the nanoscale and address spatial complexity to ensure stable and accurate signal acquisition. Finally, we consider future directions in MIP-based nanosensor development. Full article
(This article belongs to the Special Issue Recent Advances in Molecularly Imprinted-Polymer-Based Biosensors)
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21 pages, 1147 KB  
Review
Recent Advances in Developing Cell-Free Protein Synthesis Biosensors for Medical Diagnostics and Environmental Monitoring
by Tyler P. Green, Joseph P. Talley and Bradley C. Bundy
Biosensors 2025, 15(8), 499; https://doi.org/10.3390/bios15080499 - 3 Aug 2025
Cited by 15 | Viewed by 7156
Abstract
Cell-free biosensors harness the selectivity of cellular machinery without living cells’ constraints, offering advantages in environmental monitoring, medical diagnostics, and biotechnological applications. This review examines recent advances in cell-free biosensor development, highlighting their ability to detect diverse analytes including heavy metals, organic pollutants, [...] Read more.
Cell-free biosensors harness the selectivity of cellular machinery without living cells’ constraints, offering advantages in environmental monitoring, medical diagnostics, and biotechnological applications. This review examines recent advances in cell-free biosensor development, highlighting their ability to detect diverse analytes including heavy metals, organic pollutants, pathogens, and clinical biomarkers with high sensitivity and specificity. We analyze technological innovations in cell-free protein synthesis optimization, preservation strategies, and field deployment methods that have enhanced sensitivity, and practical applicability. The integration of synthetic biology approaches has enabled complex signal processing, multiplexed detection, and novel sensor designs including riboswitches, split reporter systems, and metabolic sensing modules. Emerging materials such as supported lipid bilayers, hydrogels, and artificial cells are expanding biosensor capabilities through microcompartmentalization and electronic integration. Despite significant progress, challenges remain in standardization, sample interference mitigation, and cost reduction. Future opportunities include smartphone integration, enhanced preservation methods, and hybrid sensing platforms. Cell-free biosensors hold particular promise for point-of-care diagnostics in resource-limited settings, environmental monitoring applications, and food safety testing, representing essential tools for addressing global challenges in healthcare, environmental protection, and biosecurity. Full article
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32 pages, 12213 KB  
Review
Capacitive Sensors for Label-Free Detection in High-Ionic-Strength Bodily Fluids: A Review
by Seerat Sekhon, Richard Bayford and Andreas Demosthenous
Biosensors 2025, 15(8), 491; https://doi.org/10.3390/bios15080491 - 30 Jul 2025
Cited by 10 | Viewed by 4226
Abstract
Capacitive sensors are platforms that enable label-free, real-time detection at low non-perturbing voltages. These sensors do not rely on Faradaic processes, thereby eliminating the need for redox-active species and simplifying system integration for point-of-care diagnostics. However, their sensitivity in high-ionic-strength solutions, such as [...] Read more.
Capacitive sensors are platforms that enable label-free, real-time detection at low non-perturbing voltages. These sensors do not rely on Faradaic processes, thereby eliminating the need for redox-active species and simplifying system integration for point-of-care diagnostics. However, their sensitivity in high-ionic-strength solutions, such as bodily fluids, is limited due to a reduced Debye length and non-specific interactions. The present review highlights advances in material integration, surface modification, and signal enhancement techniques to mitigate the challenges of deploying capacitive sensors in biofluids (sweat, saliva, blood, serum). This work further expands on the promise of such sensors for advancing liquid biopsies and highlights key technical challenges in translating capacitive systems to clinics. Full article
(This article belongs to the Special Issue Novel Designs and Applications for Electrochemical Biosensors)
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33 pages, 8681 KB  
Review
AI-Empowered Electrochemical Sensors for Biomedical Applications: Technological Advances and Future Challenges
by Yafeng Liu, Xiaohui Liu, Xuemei Wang and Hui Jiang
Biosensors 2025, 15(8), 487; https://doi.org/10.3390/bios15080487 - 28 Jul 2025
Cited by 41 | Viewed by 5207
Abstract
Biomarkers play a pivotal role in disease diagnosis, therapeutic efficacy evaluation, prognostic assessment, and drug screening. However, the trace concentrations of these markers in complex physiological environments pose significant challenges to efficient detection. It is necessary to avoid interference from non-specific signals, which [...] Read more.
Biomarkers play a pivotal role in disease diagnosis, therapeutic efficacy evaluation, prognostic assessment, and drug screening. However, the trace concentrations of these markers in complex physiological environments pose significant challenges to efficient detection. It is necessary to avoid interference from non-specific signals, which may lead to misjudgment of other substances as biomarkers and affect the accuracy of detection results. With the rapid advancements in electrochemical technologies and artificial intelligence (AI) algorithms, intelligent electrochemical biosensors have emerged as a promising approach for biomedical detection, offering speed, specificity, high sensitivity, and accuracy. This review focuses on elaborating the latest applications of AI-empowered electrochemical biosensors in the biomedical field, including disease diagnosis, treatment monitoring, drug development, and wearable devices. AI algorithms can further improve the accuracy, sensitivity, and repeatability of electrochemical sensors through the screening and performance prediction of sensor materials, as well as the feature extraction and noise reduction suppression of sensing signals. Even in complex physiological microenvironments, they can effectively address common issues such as electrode fouling, poor signal-to-noise ratio, chemical interference, and matrix effects. This work may provide novel insights for the development of next-generation intelligent biosensors for precision medicine. Full article
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32 pages, 3198 KB  
Review
Shining the Path of Precision Diagnostic: Advancements in Photonic Sensors for Liquid Biopsy
by Paola Colapietro, Giuseppe Brunetti, Carlotta Panciera, Aurora Elicio and Caterina Ciminelli
Biosensors 2025, 15(8), 473; https://doi.org/10.3390/bios15080473 - 22 Jul 2025
Cited by 22 | Viewed by 3096
Abstract
Liquid biopsy (LB) has gained attention as a valuable approach for cancer diagnostics, providing a minimally invasive option compared to conventional tissue biopsies and helping to overcome issues related to patient discomfort and procedural invasiveness. Recent advances in biosensor technologies, particularly photonic sensors, [...] Read more.
Liquid biopsy (LB) has gained attention as a valuable approach for cancer diagnostics, providing a minimally invasive option compared to conventional tissue biopsies and helping to overcome issues related to patient discomfort and procedural invasiveness. Recent advances in biosensor technologies, particularly photonic sensors, have improved the accuracy, speed, and real-time capabilities for detecting circulating biomarkers in biological fluids. Incorporating these tools into clinical practice facilitates more informed therapeutic choices and contributes to tailoring treatments to individual patient profiles. This review highlights the clinical potential of LB, examines technological limitations, and outlines future research directions. Departing from traditional biosensor focused reviews, it adopts a reverse-mapping approach grounded in clinically relevant tumor biomarkers. Specifically, biomarkers associated with prevalent cancers, such as breast, prostate, and lung cancers, serve as the starting point for identifying the most suitable photonic sensing platforms. The analysis underscores the need to align sensor design with the physicochemical properties of each biomarker and the operational requirements of the application. No photonic platform is universally optimal; rather, each exhibits specific strengths depending on performance metrics such as sensitivity, limit of detection, and easy system integration. Within this framework, the review provides a comprehensive assessment of emerging photonic biosensors and outlines key priorities to support their effective clinical translation in cancer diagnostics. Full article
(This article belongs to the Special Issue Lab-on-a-Chip Devices for Point-of-Care Diagnostics)
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16 pages, 3231 KB  
Article
Aptamer-Conjugated Magnetic Nanoparticles Integrated with SERS for Multiplex Salmonella Detection
by Fan Sun, Kun Pang, Keke Yang, Li Zheng, Mengmeng Wang, Yufeng Wang, Qiang Chen, Zihong Ye, Pei Liang and Xiaoping Yu
Biosensors 2025, 15(7), 464; https://doi.org/10.3390/bios15070464 - 19 Jul 2025
Cited by 7 | Viewed by 3622
Abstract
Salmonella is a rapidly spreading and widespread zoonotic infectious disease that poses a serious threat to the safety of both poultry and human lives. Therefore, the timely detection of Salmonella in foods and animals has become an urgent need for food safety. This [...] Read more.
Salmonella is a rapidly spreading and widespread zoonotic infectious disease that poses a serious threat to the safety of both poultry and human lives. Therefore, the timely detection of Salmonella in foods and animals has become an urgent need for food safety. This work describes the construction of an aptamer-based sensor for Salmonella detection, using Fe3O4 magnetic beads and Ag@Au core–shell nanoparticles-embedded 4-mercaptobenzoic acid (4MBA). Leveraging the high affinity between biotin and streptavidin, aptamers were conjugated to Fe3O4 magnetic beads. These beads were then combined with Ag@4MBA@Au nanoparticles functionalized with complementary aptamers through hydrogen bonding and π-π stacking interactions, yielding a SERS-based aptamer sensor with optimized Raman signals from 4MBA. When target bacteria are present, aptamer-conjugated magnetic beads exhibit preferential binding to the bacteria, leading to a decrease in the surface-enhanced Raman scattering (SERS) signal. And it was used for the detection of five different serotypes of Salmonella, respectively, and the results showed that the aptamer sensor exhibited a good linear relationship between the concentration range of 102–108 CFU/mL and LOD is 35.51 CFU/mL. The SERS aptasensor was utilized for the detection of spiked authentic samples with recoveries between 94.0 and 100.4%, which proved the usability of the method and helped to achieve food safety detection. Full article
(This article belongs to the Special Issue Aptamer-Based Sensing: Designs and Applications)
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11 pages, 1778 KB  
Communication
Ultra-Sensitive Detection of Chloramphenicol by CdS@NiMoS Nanorods-Based Photoelectrochemical Aptasensor
by Hebin Sun, Yimeng Sun, Tong Qi, Zhenyu Wang, Jianlong Zhao and Lijuan Liang
Biosensors 2025, 15(7), 454; https://doi.org/10.3390/bios15070454 - 14 Jul 2025
Cited by 3 | Viewed by 1621
Abstract
A novel nanomaterial photoelectrochemical aptamer sensor based on CdS@NiMoS heterojunction nanocomposites was constructed for highly sensitive detection of chloramphenicol (CAP) in antibiotic residues. Through optimization of the material synthesis process, the optimal doping ratio of MoS2 to Ni3+ (70% MoS2 [...] Read more.
A novel nanomaterial photoelectrochemical aptamer sensor based on CdS@NiMoS heterojunction nanocomposites was constructed for highly sensitive detection of chloramphenicol (CAP) in antibiotic residues. Through optimization of the material synthesis process, the optimal doping ratio of MoS2 to Ni3+ (70% MoS2 and 10% Ni3+) was identified, which significantly enhanced the photogenerated carrier separation efficiency. In thin-film preparation, comparative analysis of four film-forming methods led to the determination of an optimal process with stability. To achieve highly specific CAP detection, the nanocomposite chip was integrated with nucleic acid aptamer biorecognition elements within a standard three-electrode detection system. Experimental results demonstrated a linear response (R2 = 0.998) in the 0.1–2 μM concentration range, with a detection limit of 3.69 nM (3σ/S). Full article
(This article belongs to the Special Issue Nanotechnology Biosensing in Bioanalysis and Beyond)
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40 pages, 2353 KB  
Review
Electrochemical Impedance Spectroscopy-Based Biosensors for Label-Free Detection of Pathogens
by Huaiwei Zhang, Zhuang Sun, Kaiqiang Sun, Quanwang Liu, Wubo Chu, Li Fu, Dan Dai, Zhiqiang Liang and Cheng-Te Lin
Biosensors 2025, 15(7), 443; https://doi.org/10.3390/bios15070443 - 10 Jul 2025
Cited by 73 | Viewed by 10811
Abstract
The escalating threat of infectious diseases necessitates the development of diagnostic technologies that are not only rapid and sensitive but also deployable at the point of care. Electrochemical impedance spectroscopy (EIS) has emerged as a leading technique for the label-free detection of pathogens, [...] Read more.
The escalating threat of infectious diseases necessitates the development of diagnostic technologies that are not only rapid and sensitive but also deployable at the point of care. Electrochemical impedance spectroscopy (EIS) has emerged as a leading technique for the label-free detection of pathogens, offering a unique combination of sensitivity, non-invasiveness, and adaptability. This review provides a comprehensive overview of the design and application of EIS-based biosensors tailored for pathogen detection, focusing on critical components such as biorecognition elements, electrode materials, nanomaterial integration, and surface immobilization strategies. Special emphasis is placed on the mechanisms of signal generation under Faradaic and non-Faradaic modes and how these underpin performance characteristics such as the limit of detection, specificity, and response time. The application spectrum spans bacterial, viral, fungal, and parasitic pathogens, with case studies highlighting detection in complex matrices such as blood, saliva, food, and environmental water. Furthermore, integration with microfluidics and point-of-care systems is explored as a pathway toward real-world deployment. Emerging strategies for multiplexed detection and the utilization of novel nanomaterials underscore the dynamic evolution of the field. Key challenges—including non-specific binding, matrix effects, the inherently low ΔRct/decade sensitivity of impedance transduction, and long-term stability—are critically evaluated alongside recent breakthroughs. This synthesis aims to support the future development of robust, scalable, and user-friendly EIS-based pathogen biosensors with the potential to transform diagnostics across healthcare, food safety, and environmental monitoring. Full article
(This article belongs to the Special Issue Material-Based Biosensors and Biosensing Strategies)
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38 pages, 3752 KB  
Review
Recent Advances in Metal–Organic Framework-Based Nanozymes for Intelligent Microbial Biosensing: A Comprehensive Review of Biomedical and Environmental Applications
by Alemayehu Kidanemariam and Sungbo Cho
Biosensors 2025, 15(7), 437; https://doi.org/10.3390/bios15070437 - 7 Jul 2025
Cited by 25 | Viewed by 6352
Abstract
Metal–organic framework (MOF)-based nanozymes represent a groundbreaking frontier in advanced microbial biosensing, offering unparalleled catalytic precision and structural tunability to mimic natural enzymes with superior stability and specificity. By engineering the structural features and forming composites, MOFs are precisely tailored to amplify nanozymatic [...] Read more.
Metal–organic framework (MOF)-based nanozymes represent a groundbreaking frontier in advanced microbial biosensing, offering unparalleled catalytic precision and structural tunability to mimic natural enzymes with superior stability and specificity. By engineering the structural features and forming composites, MOFs are precisely tailored to amplify nanozymatic activity, enabling the highly sensitive, rapid, and cost-effective detection of a broad spectrum of microbial pathogens critical to biomedical diagnostics and environmental monitoring. These advanced biosensors surpass traditional enzyme systems in robustness and reusability, integrating seamlessly with smart diagnostic platforms for real-time, on-site microbial identification. This review highlights cutting-edge developments in MOF nanozyme design, composite engineering, and signal transduction integration while addressing pivotal challenges such as biocompatibility, complex matrix interference, and scalable manufacturing. Looking ahead, the convergence of multifunctional MOF nanozymes with portable technologies and optimized in vivo performance will drive transformative breakthroughs in early disease detection, antimicrobial resistance surveillance, and environmental pathogen control, establishing a new paradigm in next-generation smart biosensing. Full article
(This article belongs to the Special Issue Microbial Biosensor: From Design to Applications—2nd Edition)
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31 pages, 1686 KB  
Review
Strategic Detection of Escherichia coli in the Poultry Industry: Food Safety Challenges, One Health Approaches, and Advances in Biosensor Technologies
by Jacquline Risalvato, Alaa H. Sewid, Shigetoshi Eda, Richard W. Gerhold and Jie Jayne Wu
Biosensors 2025, 15(7), 419; https://doi.org/10.3390/bios15070419 - 1 Jul 2025
Cited by 20 | Viewed by 6315
Abstract
Escherichia coli (E. coli) remains a major concern in poultry production due to its ability to incite foodborne illness and public health crisis, zoonotic potential, and the increasing prevalence of antibiotic-resistant strains. The contamination of poultry products with pathogenic E. coli [...] Read more.
Escherichia coli (E. coli) remains a major concern in poultry production due to its ability to incite foodborne illness and public health crisis, zoonotic potential, and the increasing prevalence of antibiotic-resistant strains. The contamination of poultry products with pathogenic E. coli, including avian pathogenic E. coli (APEC) and Shiga toxin-producing E. coli (STEC), presents risks at multiple stages of the poultry production cycle. The stages affected by E. coli range from, but are not limited to, the hatcheries to grow-out operations, slaughterhouses, and retail markets. While traditional detection methods such as culture-based assays and polymerase chain reaction (PCR) are well-established for E. coli detection in the food supply chain, their time, cost, and high infrastructure demands limit their suitability for rapid and field-based surveillance—hindering the ability for effective cessation and handling of outbreaks. Biosensors have emerged as powerful diagnostic tools that offer rapid, sensitive, and cost-effective alternatives for E. coli detection across various stages of poultry development and processing where detection is needed. This review examines current biosensor technologies designed to detect bacterial biomarkers, toxins, antibiotic resistance genes, and host immune response indicators for E. coli. Emphasis is placed on field-deployable and point-of-care (POC) platforms capable of integrating into poultry production environments. In addition to enhancing early pathogen detection, biosensors support antimicrobial resistance monitoring, facilitate integration into Hazard Analysis Critical Control Points (HACCP) systems, and align with the One Health framework by improving both animal and public health outcomes. Their strategic implementation in slaughterhouse quality control and marketplace testing can significantly reduce contamination risk and strengthen traceability in the poultry value chain. As biosensor technology continues to evolve, its application in E. coli surveillance is poised to play a transformative role in sustainable poultry production and global food safety. Full article
(This article belongs to the Special Issue Biosensors for Food Safety)
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23 pages, 2366 KB  
Article
Whole-Cell Fiber-Optic Biosensor for Real-Time, On-Site Sediment and Water Toxicity Assessment: Applications at Contaminated Sites Across Israel
by Gal Carmeli, Abraham Abbey Paul, Kathelina Kristollari, Evgeni Eltzov, Albert Batushansky and Robert S. Marks
Biosensors 2025, 15(7), 404; https://doi.org/10.3390/bios15070404 - 22 Jun 2025
Cited by 4 | Viewed by 4787
Abstract
Sediments are key players in the optimum functioning of ecosystems; however, they also represent the largest known repository of harmful contaminants. The vast variety of these sediment-associated contaminants may exert harmful effects on marine communities and can impair ecosystem functioning. Whole-cell biosensors are [...] Read more.
Sediments are key players in the optimum functioning of ecosystems; however, they also represent the largest known repository of harmful contaminants. The vast variety of these sediment-associated contaminants may exert harmful effects on marine communities and can impair ecosystem functioning. Whole-cell biosensors are a rapid and biologically relevant tool for assessing environmental toxicity. Therefore, in this study, we developed a bioassay-based toxicity measurement system using genetically modified bacteria to create a whole-cell optical biosensor. Briefly, reporter bacteria were integrated and immobilized using a calcium alginate matrix on fiber-optic tips connected to a photon counter placed inside a light-proof, portable case. The calcium alginate matrix acts as a semi-permeable membrane that protects the reporter-encapsulated optical fiber tips and allows the inward passage of toxicant(s) to induce a dose-dependent response in the bioreporter. The samples were tested by directly submerging the fiber tip with immobilized bacteria into vials containing either water or suspended sediment samples, and the subsequent bioluminescent responses were acquired. In addition to bioavailable sediment toxicity assessments, conventional chemical methods, such as liquid chromatography–mass spectroscopy (LC-MS) and inductively coupled plasma optical emission spectroscopy (ICP-OES), were used for comprehensive evaluation. The results demonstrated the efficacy of the biosensor in detecting various toxicity levels corresponding to identified contaminants, highlighting its potential integration into environmental monitoring frameworks for enhanced sediment and water quality assessments. Despite its utility, this study notes the system’s operational challenges in field conditions, recommending future enhancements for improved portability and usability in remote locations. Full article
(This article belongs to the Special Issue Optical Biosensors for Environmental Monitoring)
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44 pages, 18112 KB  
Review
Plasmonic and Dielectric Metasurfaces for Enhanced Spectroscopic Techniques
by Borja García García, María Gabriela Fernández-Manteca, Dimitrios C. Zografopoulos, Celia Gómez-Galdós, Alain A. Ocampo-Sosa, Luis Rodríguez-Cobo, José Francisco Algorri and Adolfo Cobo
Biosensors 2025, 15(7), 401; https://doi.org/10.3390/bios15070401 - 20 Jun 2025
Cited by 16 | Viewed by 6763
Abstract
Spectroscopic techniques such as Surface-Enhanced Raman Scattering (SERS), Surface-Enhanced Infrared Absorption (SEIRA), and Surface-Enhanced Fluorescence (SEF) are essential analytical techniques used to study the composition of materials by analyzing the way materials scatter light, absorb infrared radiation or emit fluorescence signals. This provides [...] Read more.
Spectroscopic techniques such as Surface-Enhanced Raman Scattering (SERS), Surface-Enhanced Infrared Absorption (SEIRA), and Surface-Enhanced Fluorescence (SEF) are essential analytical techniques used to study the composition of materials by analyzing the way materials scatter light, absorb infrared radiation or emit fluorescence signals. This provides information about their molecular structure and properties. However, traditional SERS, SEIRA, and SEF techniques can be limited in sensitivity, resolution, and reproducibility, hindering their ability to detect and analyze trace amounts of substances or complex molecular structures. Metasurfaces, a class of engineered two-dimensional metamaterials with unique optical properties, have emerged as a promising tool to overcome these limitations and enhance spectroscopic techniques. This article provides a state-of-the-art overview of metasurfaces for enhanced SERS, SEIRA and SEF, covering their theoretical background, different types, advantages, disadvantages, and potential applications. Full article
(This article belongs to the Special Issue Photonics for Bioapplications: Sensors and Technology—2nd Edition)
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33 pages, 4455 KB  
Review
Small Toxic Molecule Detection and Elimination Using Molecularly Imprinted Polymers (MIPs)
by Min Seok Kang, Jin-Ho Lee and Ki Su Kim
Biosensors 2025, 15(6), 393; https://doi.org/10.3390/bios15060393 - 18 Jun 2025
Cited by 22 | Viewed by 4791
Abstract
Molecularly imprinted polymers (MIPs) provide selective, robust, and cost-effective platforms for the detection and removal of small toxic molecules in environmental, food, and biomedical contexts. This review offers a comprehensive overview of recent advancements in MIP-based systems, emphasizing critical design factors such as [...] Read more.
Molecularly imprinted polymers (MIPs) provide selective, robust, and cost-effective platforms for the detection and removal of small toxic molecules in environmental, food, and biomedical contexts. This review offers a comprehensive overview of recent advancements in MIP-based systems, emphasizing critical design factors such as template selection, functional monomers, polymerization methods, and binding kinetics. The impact of these parameters on improving sensitivity, selectivity, and reusability is thoroughly examined. Additionally, current advantages, limitations, and enduring challenges are addressed. By highlighting emerging strategies and interdisciplinary innovations, this work aims to guide the development of more efficient and sustainable technologies for small-molecule toxin detection and remediation. Full article
(This article belongs to the Special Issue Molecularly Imprinted Polymers-Based Biosensors)
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13 pages, 1792 KB  
Article
A High-Sensitivity, Bluetooth-Enabled PCB Biosensor for HER2 and CA15-3 Protein Detection in Saliva: A Rapid, Non-Invasive Approach to Breast Cancer Screening
by Hsiao-Hsuan Wan, Chao-Ching Chiang, Fan Ren, Cheng-Tse Tsai, Yu-Siang Chou, Chun-Wei Chiu, Yu-Te Liao, Dan Neal, Coy D. Heldermon, Mateus G. Rocha and Josephine F. Esquivel-Upshaw
Biosensors 2025, 15(6), 386; https://doi.org/10.3390/bios15060386 - 15 Jun 2025
Cited by 7 | Viewed by 4991
Abstract
Breast cancer is a leading cause of cancer-related mortality worldwide, requiring efficient diagnostic tools for early detection and monitoring. Human epidermal growth factor receptor 2 (HER2) is a key biomarker for breast cancer classification, typically assessed using immunohistochemistry (IHC). However, IHC requires invasive [...] Read more.
Breast cancer is a leading cause of cancer-related mortality worldwide, requiring efficient diagnostic tools for early detection and monitoring. Human epidermal growth factor receptor 2 (HER2) is a key biomarker for breast cancer classification, typically assessed using immunohistochemistry (IHC). However, IHC requires invasive biopsies and time-intensive laboratory procedures. In this study, we present a biosensor integrated with a reusable printed circuit board (PCB) and functionalized glucose test strips designed for rapid and non-invasive HER2 detection in saliva. The biosensor achieved a limit of detection of 10−15 g/mL, 4 to 5 orders of magnitude more sensitive than the enzyme-linked immunosorbent assay (ELISA), with a sensitivity of 95/dec and a response time of 1 s. In addition to HER2, the biosensor also detects cancer antigen 15-3 (CA15-3), another clinically relevant breast cancer biomarker. The CA15-3 test demonstrated an equally low limit of detection, 10−15 g/mL, and a higher sensitivity, 190/dec, further validated using human saliva samples. Clinical validation using 29 saliva samples confirmed our biosensor’s ability to distinguish between healthy, in situ breast cancer, and invasive breast cancer patients. The system, which integrates a Bluetooth Low-Energy (BLE) module, enables remote monitoring, reduces hospital visits, and enhances accessibility for point-of-care and mobile screening applications. This ultra-sensitive, rapid, and portable biosensor can serve as a promising alternative for breast cancer detection and monitoring, particularly in rural and underserved communities. Full article
(This article belongs to the Special Issue Aptamer-Based Biosensors for Point-of-Care Diagnostics)
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16 pages, 3644 KB  
Article
Sensing Protein Structural Transitions with Microfluidic Modulation Infrared Spectroscopy
by Lathan Lucas, Phoebe S. Tsoi, Ananya Nair, Allan Chris M. Ferreon and Josephine C. Ferreon
Biosensors 2025, 15(6), 382; https://doi.org/10.3390/bios15060382 - 13 Jun 2025
Cited by 5 | Viewed by 2874
Abstract
Microfluidic modulation spectroscopy-infrared (MMS) offers a label-free, high-sensitivity approach for quantifying changes in protein secondary structures under native solution conditions. MMS subtracts the solvent backgrounds from sample signals by alternately flowing proteins and matched buffers through a microfluidic chamber, yielding clear amide I [...] Read more.
Microfluidic modulation spectroscopy-infrared (MMS) offers a label-free, high-sensitivity approach for quantifying changes in protein secondary structures under native solution conditions. MMS subtracts the solvent backgrounds from sample signals by alternately flowing proteins and matched buffers through a microfluidic chamber, yielding clear amide I spectra from microliter volumes. In this study, we validated MMS on canonical globular proteins, bovine serum albumin, mCherry, and lysozyme, demonstrating accurate detection and resolution of α-helix, β-sheet, and mixed-fold structures. Applying MMS to the intrinsically disordered protein Tau, we detected environment-driven shifts in transient conformers: both the acidic (pH 2.5) and alkaline (pH 10) conditions increased the turn/unordered structures and decreased the α-helix content relative to the neutral pH, highlighting the charge-mediated destabilization of the labile motifs. Hyperphosphorylation of Tau yielded a modest decrease in the α-helical fraction and an increase in the turn/unordered structures. Comparison of monomeric and aggregated hyperphosphorylated Tau revealed a dramatic gain in β-sheet and a loss in turn/unordered structures upon amyloid fibril formation, confirming MMS’s ability to distinguish disordered monomers from amyloids. These findings establish MMS as a robust platform for detecting protein secondary structures and monitoring aggregation pathways in both folded and disordered systems. The sensitive detection of structural transitions offers opportunities for probing misfolding mechanisms and advancing our understanding of aggregation-related diseases. Full article
(This article belongs to the Special Issue Design and Application of Microfluidic Biosensors in Biomedicine)
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36 pages, 3955 KB  
Review
Electrochemical Microneedles for Real-Time Monitoring in Interstitial Fluid: Emerging Technologies and Future Directions
by Suhyeon Cha, Min Yu Choi, Min Jung Kim, Sang Baek Sim, Izzati Haizan and Jin-Ha Choi
Biosensors 2025, 15(6), 380; https://doi.org/10.3390/bios15060380 - 12 Jun 2025
Cited by 22 | Viewed by 9829
Abstract
Conventional blood-based detection methods for biomarkers and analytes face significant limitations, including complex processing, variability in blood components, and the inability to provide continuous monitoring. These challenges hinder the early diagnosis and effective management of various health conditions. Electrochemical microneedles (MNs) have emerged [...] Read more.
Conventional blood-based detection methods for biomarkers and analytes face significant limitations, including complex processing, variability in blood components, and the inability to provide continuous monitoring. These challenges hinder the early diagnosis and effective management of various health conditions. Electrochemical microneedles (MNs) have emerged as a minimally invasive and highly efficient platform to overcome these barriers, enabling continuous molecular monitoring by directly accessing the interstitial fluid. Electrochemical MNs offer several advantages, including reduced patient discomfort, real-time data acquisition, enhanced specificity, and potential applications in wearable, long-term monitoring. In this review, we first analyze material selection and fabrication techniques to optimize sensor performance, stability, and biocompatibility. We then examine diverse detection strategies utilized in electrochemical MNs, including enzyme-based, aptamer-based, and antibody-based sensing mechanisms, each offering unique benefits in sensitivity and selectivity. Finally, we highlight the integration of electrochemical MN technology with multi-target detection, AI-driven analytics, and theragnostic capabilities. This convergence offers strong potential for smart healthcare and precision medicine. Through these technological innovations, electrochemical MNs are expected to play an important role in advancing continuous, noninvasive health monitoring and personalized medical care. Full article
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22 pages, 1476 KB  
Article
Wearable Ultrasound-Imaging-Based Visual Feedback (UVF) Training for Ankle Rehabilitation of Chronic Stroke Survivors: A Proof-of-Concept Randomized Crossover Study
by Yu-Yan Luo, Chen Huang, Zhen Song, Vaheh Nazari, Arnold Yu-Lok Wong, Lin Yang, Mingjie Dong, Mingming Zhang, Yong-Ping Zheng, Amy Siu-Ngor Fu and Christina Zong-Hao Ma
Biosensors 2025, 15(6), 365; https://doi.org/10.3390/bios15060365 - 6 Jun 2025
Cited by 5 | Viewed by 3342
Abstract
This study investigated the effect of wearable ultrasound-imaging-based visual feedback (UVF) on assisting paretic ankle dorsiflexion training of chronic stroke survivors. Thirty-three participants with unilateral hemiplegia performed maximal isometric contractions on an isokinetic dynamometer in randomized conditions with and without UVF that provided [...] Read more.
This study investigated the effect of wearable ultrasound-imaging-based visual feedback (UVF) on assisting paretic ankle dorsiflexion training of chronic stroke survivors. Thirty-three participants with unilateral hemiplegia performed maximal isometric contractions on an isokinetic dynamometer in randomized conditions with and without UVF that provided by a wearable ultrasound imaging system. Torque parameters (mean, peak, percentage of maximal voluntary contraction) and tibialis anterior muscle thickness were analyzed across different contraction phases. Statistical comparisons were conducted using paired t-tests or Wilcoxon tests. Correlation analyses were performed using Pearson’s or Spearman’s tests. Results demonstrated that UVF significantly improved torque output, as evidence by the increased percentage of maximal voluntary contraction (%MVC) during entire contractions (p = 0.007), increased mean (p ≤ 0.022) and peak (p ≤ 0.044) torque and the %MVC (p ≤ 0.004) during mid and end phases, and larger muscle thickness during mid contraction (p = 0.045). Moderate correlations were found between torque and muscle thickness (r ≥ 0.30, p ≤ 0.049). These findings preliminarily supported the positive outcomes of real-time wearable UVFs in enhancing paretic ankle dorsiflexion strength and force control during isometric contractions in chronic stroke survivors. While the developed and validated new training protocol may potentially serve as a practical adjunct to existing rehabilitation approaches, further investigations emphasizing the functional outcomes and clinical translations are still needed to verify the clinical utility. Full article
(This article belongs to the Special Issue Innovative Biosensing Technologies for Sustainable Healthcare)
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32 pages, 5494 KB  
Review
Colorimetric Biosensors: Advancements in Nanomaterials and Cutting-Edge Detection Strategies
by Yubeen Lee, Izzati Haizan, Sang Baek Sim and Jin-Ha Choi
Biosensors 2025, 15(6), 362; https://doi.org/10.3390/bios15060362 - 5 Jun 2025
Cited by 47 | Viewed by 8391
Abstract
Colorimetric-based biosensors are practical detection devices that can detect the presence and concentration of biomarkers through simple color changes. Conventional laboratory-based tests are highly sensitive but require long processing times and expensive equipment, which makes them difficult to apply for on-site diagnostics. In [...] Read more.
Colorimetric-based biosensors are practical detection devices that can detect the presence and concentration of biomarkers through simple color changes. Conventional laboratory-based tests are highly sensitive but require long processing times and expensive equipment, which makes them difficult to apply for on-site diagnostics. In contrast, the colorimetric method offers advantages for point-of-care testing and real-time monitoring due to its flexibility, simple operation, rapid results, and versatility across many applications. In order to enhance the color change reactions in colorimetric techniques, functional nanomaterials are often integrated due to their desirable intrinsic properties. In this review, the working principles of nanomaterial-based detection strategies in colorimetric systems are introduced. In addition, current signal amplification methods for colorimetric biosensors are comprehensively outlined and evaluated. Finally, the latest trends in artificial intelligence (AI) and machine learning integration into colorimetric-based biosensors, including their potential for technological advancements in the near future, are discussed. Future research is expected to develop highly sensitive and multifunctional colorimetric methods, which will serve as powerful alternatives for point-of-care testing and self-testing. Full article
(This article belongs to the Special Issue Functional Materials for Biosensing Applications)
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37 pages, 9912 KB  
Review
Advances in Hydrogel-Integrated SERS Platforms: Innovations, Applications, Challenges, and Future Prospects in Food Safety Detection
by Xorlali Nunekpeku, Huanhuan Li, Ayesha Zahid, Chenhui Li and Wei Zhang
Biosensors 2025, 15(6), 363; https://doi.org/10.3390/bios15060363 - 5 Jun 2025
Cited by 20 | Viewed by 5693
Abstract
Background: Food safety remains a global concern due to biological and chemical contaminants, including adulterants, pathogens, antibiotic residues, and pesticides. Traditional detection methods are accurate but limited by time requirements, complex sample preparation, high costs, and poor field applicability. Surface-Enhanced Raman Spectroscopy [...] Read more.
Background: Food safety remains a global concern due to biological and chemical contaminants, including adulterants, pathogens, antibiotic residues, and pesticides. Traditional detection methods are accurate but limited by time requirements, complex sample preparation, high costs, and poor field applicability. Surface-Enhanced Raman Spectroscopy (SERS) offers non-destructive analysis with low detection limits and high specificity, yet conventional SERS substrates face challenges with reproducibility, nanoparticle aggregation, and sensitivity in food matrices. Hydrogels have emerged as supporting materials for SERS due to their water content, tunable porosity, flexibility, and ability to entrap plasmonic nanostructures. Scope and Approach: This review examines recent advances in hydrogel-integrated SERS platforms for food safety applications. The three-dimensional structure of hydrogels enables homogeneous distribution of metal nanoparticles, prevents aggregation, and offers analyte enrichment. We analyze material design, functionalization strategies, and how hydrogel properties—crosslinking density, porosity, surface charge, and nanoparticle distribution—influence SERS performance in food matrices. Key Findings and Conclusions: Hydrogel-integrated SERS platforms demonstrate superior performance in detecting various food contaminants—including pesticides, adulterants, and additives—in real food matrices, often achieving detection limits in the nanomolar to picomolar range, depending on the analyte and substrate design. Current limitations include storage stability concerns, batch-to-batch variability, and regulatory acceptance hurdles. Future research directions should focus on multiplex detection capabilities, integration with smart sensing technologies, and industrial scalability to facilitate practical deployment in global food safety monitoring across diverse supply chains. Full article
(This article belongs to the Special Issue Advanced SERS Biosensors for Detection and Analysis)
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31 pages, 4568 KB  
Review
Stimuli-Responsive DNA Hydrogel Design Strategies for Biomedical Applications
by Minhyuk Lee, Minjae Lee, Sungjee Kim and Nokyoung Park
Biosensors 2025, 15(6), 355; https://doi.org/10.3390/bios15060355 - 4 Jun 2025
Cited by 17 | Viewed by 5862
Abstract
Hydrogels are three-dimensional network structures composed of hydrophilic polymers that can swell in water and are very similar to soft tissues such as connective tissue or the extracellular matrix. DNA hydrogels are particularly notable for biomedical applications due to their high biocompatibility, physiological [...] Read more.
Hydrogels are three-dimensional network structures composed of hydrophilic polymers that can swell in water and are very similar to soft tissues such as connective tissue or the extracellular matrix. DNA hydrogels are particularly notable for biomedical applications due to their high biocompatibility, physiological stability, molecular recognition, biodegradability, easy functionalization, and low immunogenicity. Based on these advantages, stimuli-responsive DNA hydrogels that have the property of reversibly changing their structure in response to various microenvironments or molecules are attracting attention as smart nanomaterials that can be applied to biosensing and material transfer, such as in the case of cells and drugs. As DNA nanotechnology advances, DNA can be hybridized with a variety of nanomaterials, from inorganic nanomaterials such as gold nanoparticles (AuNPs) and quantum dots (QDs) to synthetic polymers such as polyacrylamide (PAAm) and poly(N-isopropylacrylamide) (pNIPAM). These hybrid structures exhibit various optical and chemical properties. This review discusses recent advances and remaining challenges in biomedical applications of stimuli-responsive smart DNA hydrogel-based systems. It also highlights various types of hybridized DNA hydrogel, explores various response mechanism strategies of stimuli-responsive DNA hydrogel, and provides insights and prospects for biomedical applications such as biosensing and drug delivery. Full article
(This article belongs to the Special Issue Hydrogel-Based Biosensors: From Design to Applications)
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11 pages, 1139 KB  
Article
Electrochemical Sensor Platform for Rapid Detection of Foodborne Toxins
by Kundan Kumar Mishra, Krupa M. Thakkar, Vikram Narayanan Dhamu, Sriram Muthukumar and Shalini Prasad
Biosensors 2025, 15(6), 361; https://doi.org/10.3390/bios15060361 - 4 Jun 2025
Cited by 6 | Viewed by 2337
Abstract
Zearalenone (ZEA), a potent mycotoxin commonly found in contaminated grains, presents a serious threat to food safety and public health. Conventional detection methods, including culture-based assays and laboratory-bound analytical tools, are often time-consuming, require specialized infrastructure, and lack portability, limiting their utility for [...] Read more.
Zearalenone (ZEA), a potent mycotoxin commonly found in contaminated grains, presents a serious threat to food safety and public health. Conventional detection methods, including culture-based assays and laboratory-bound analytical tools, are often time-consuming, require specialized infrastructure, and lack portability, limiting their utility for rapid, on-site screening. In response, this study introduces a compact, real-time electrochemical sensing platform for the swift and selective detection of ZEA in corn flour matrices. Utilizing a non-faradaic, label-free approach based on Electrochemical Impedance Spectroscopy (EIS), the sensor leverages ZEA-specific antibodies to achieve rapid detection within 5 min. The platform demonstrates a low detection limit of 0.05 ng/mL, with a broad dynamic range from 0.1 ng/mL to 25.6 ng/mL. Reproducibility tests confirm consistent performance, with both inter- and intra-assay variation remaining under a 20% coefficient of variation (%CV). Comparative evaluation with standard benchtop systems underscores its accuracy and field applicability. This portable and user-friendly device provides a powerful tool for real-time mycotoxin monitoring, offering significant potential for improving food safety practices and enabling point-of-need testing in resource-limited settings. Full article
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29 pages, 7096 KB  
Review
Recent Progress on the Application of Microneedles for In Situ Sampling in Surface-Enhanced Raman Scattering Detection
by Weiqing Yang, Ying Chen, Xingliang Cheng, Shuojiang Liu, Huiqi Zhu and Yuling Hu
Biosensors 2025, 15(6), 350; https://doi.org/10.3390/bios15060350 - 1 Jun 2025
Cited by 7 | Viewed by 4386
Abstract
The efficient and non-invasive collection of biological samples has become a critical challenge for the continued development of surface-enhanced Raman scattering (SERS). When integrated with minimally invasive microneedle (MN) sampling technology, SERS enhances its applicability in real-time, non-invasive molecular detection. This review focuses [...] Read more.
The efficient and non-invasive collection of biological samples has become a critical challenge for the continued development of surface-enhanced Raman scattering (SERS). When integrated with minimally invasive microneedle (MN) sampling technology, SERS enhances its applicability in real-time, non-invasive molecular detection. This review focuses on the latest advances in MN-based SERS sensors. Firstly, a comprehensive summary is presented of MN types and research progress in the design and engineering of SERS-active MNs. Then, the sampling method of SERS MNs and the MN-based SERS detection mode are also described in detail. Finally, the applications of SERS MNs in fields such as disease diagnosis, drug monitoring, and food safety are highlighted. Additionally, current challenges are discussed and future development prospects are prospected with the aim of contributing to the design of MN-based SERS sensors for diverse applications. Full article
(This article belongs to the Special Issue Surface-Enhanced Raman Scattering in Biosensing Applications)
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16 pages, 2275 KB  
Article
Sweat-Sensing Patches with Integrated Hydrogel Interface for Resting Sweat Collection and Multi-Information Detection
by Lei Lu, Qiang Sun, Zihao Lin, Wenjie Xu, Xiangnan Li, Tian Wang, Yiming Lu, Huaping Wu, Lin Cheng and Aiping Liu
Biosensors 2025, 15(6), 342; https://doi.org/10.3390/bios15060342 - 29 May 2025
Cited by 13 | Viewed by 7100
Abstract
Sweat analysis represents an emerging non-invasive approach for health monitoring, yet its practical application is hindered by challenges such as insufficient natural sweat secretion and inefficient collection. To overcome these limitations, this study develops a hydrogel sheet composed of agarose and glycerol, which [...] Read more.
Sweat analysis represents an emerging non-invasive approach for health monitoring, yet its practical application is hindered by challenges such as insufficient natural sweat secretion and inefficient collection. To overcome these limitations, this study develops a hydrogel sheet composed of agarose and glycerol, which efficiently facilitates resting sweat collection without external stimulation when integrated into the microfluidic channels of a sweat-sensing patch. The microfluidic sweat-sensing patch, fabricated with laser-cut technology, features a sandwich structure that enables the measurement of sweat rate and chloride ion concentration while minimizing interference from electrochemical reactions. Additionally, a colorimetric module utilizing glucose oxidase and peroxidase is also integrated into the platform for cost-effective and efficient glucose detection through a color change that can be quantified via RGB analysis. The hydrogel interface, characterized by its optimal thickness and water content, exhibits superior absorption capability for efficient sweat collection and retention, with a negligible effect on the dilution of sweat components. This hydrogel-interfaced microfluidic platform demonstrates high efficiency in sweat collection and multi-biomarker analysis, offering a non-invasive, real-time solution for health monitoring. Its low-cost and wearable design highlights its potential for broad applications in personalized healthcare. Full article
(This article belongs to the Section Wearable Biosensors)
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40 pages, 3743 KB  
Review
Droplet Generation and Manipulation in Microfluidics: A Comprehensive Overview of Passive and Active Strategies
by Andrea Fergola, Alberto Ballesio, Francesca Frascella, Lucia Napione, Matteo Cocuzza and Simone Luigi Marasso
Biosensors 2025, 15(6), 345; https://doi.org/10.3390/bios15060345 - 29 May 2025
Cited by 44 | Viewed by 11361
Abstract
Droplet-based microfluidics (DBM) has emerged as a powerful tool for a wide range of biochemical applications, from single-cell analysis and drug screening to diagnostics and tissue engineering. This review provides a comprehensive overview of the latest advancements in droplet generation and trapping techniques, [...] Read more.
Droplet-based microfluidics (DBM) has emerged as a powerful tool for a wide range of biochemical applications, from single-cell analysis and drug screening to diagnostics and tissue engineering. This review provides a comprehensive overview of the latest advancements in droplet generation and trapping techniques, highlighting both passive and active approaches. Passive methods—such as co-flow, cross-flow, and flow-focusing geometries—rely on hydrodynamic instabilities and capillary effects, offering simplicity and integration with compact devices, though often at the cost of tunability. In contrast, active methods exploit external fields—electric, magnetic, thermal, or mechanical—to enable on-demand droplet control, allowing for higher precision and throughput. Furthermore, we explore innovative trapping mechanisms such as hydrodynamic resistance networks, microfabricated U-shaped wells, and anchor-based systems that enable precise spatial immobilization of droplets. In the final section, we also examine active droplet sorting strategies, including electric, magnetic, acoustic, and thermal methods, as essential tools for downstream analysis and high-throughput workflows. These manipulation strategies facilitate in situ chemical and biological analyses, enhance experimental reproducibility, and are increasingly adaptable to industrial-scale applications. Emphasis is placed on the design flexibility, scalability, and biological compatibility of each method, offering critical insights for selecting appropriate techniques based on experimental needs and operational constraints. Full article
(This article belongs to the Special Issue Micro/Nanofluidic System-Based Biosensors)
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13 pages, 1519 KB  
Article
Multiplexed CRISPR Assay for Amplification-Free Detection of miRNAs
by P. I. Thilini De Silva, Keshani Hiniduma, Rachelle Canete, Ketki S. Bhalerao, Sherif M. Shawky, Hansana Gunathilaka, Jessica L. Rouge, Islam M. Mosa, David C. Steffens, Kevin Manning, Breno S. Diniz and James F. Rusling
Biosensors 2025, 15(6), 346; https://doi.org/10.3390/bios15060346 - 29 May 2025
Cited by 8 | Viewed by 4265
Abstract
CRISPR-Cas proteins from bacteria are powerful tools for gene editing and molecular diagnostics. Expanding capacity of CRISPR to low cost, multiplexed assays of biomarkers is a key to future disease diagnostics, since multiple biomarker detection is essential for reliable diagnostics. Herein we describe [...] Read more.
CRISPR-Cas proteins from bacteria are powerful tools for gene editing and molecular diagnostics. Expanding capacity of CRISPR to low cost, multiplexed assays of biomarkers is a key to future disease diagnostics, since multiple biomarker detection is essential for reliable diagnostics. Herein we describe a multiplexed assay in a 3D-printed 96-well plate with CRISPR-Cas13a immobilized in each well to target three circulating blood biomarker microRNAs (miRNAs 34c-5p, 200c-3p, and 30e-5p) for Alzheimer’s disease (ALZ). Immobilized Cas13a is equipped with different crRNAs complementary to each miRNA target. MiRNA binding to crRNA complements activates the collateral RNase activity of Cas13a, cleaving a quenched fluorescent reporter (RNaseAlert) with fluorophore and quencher connected by an RNA oligonucleotide to enable fluorescence measurements. We achieved ultralow limits of detection (LOD) of 0.74 fg/mL for miRNA 34c-5p, 0.70 fg/mL for miRNA 30e-5p, and 7.4 fg/mL for miRNA 200c-3p, with dynamic ranges from LODs up to about 1800 pg/mL. The accuracy of the assay was validated by spike-recovery studies and good correlation of levels of patient plasma samples vs. a referee method. This new approach provides selective, sensitive multiplex miRNA biosensing, and simultaneously accommodates analysis of standards and controls. Full article
(This article belongs to the Special Issue Biosensors for Monitoring and Diagnostics)
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45 pages, 15218 KB  
Review
Comprehensive Analysis of Advancement in Optical Biosensing Techniques for Early Detection of Cancerous Cells
by Ayushman Ramola, Amit Kumar Shakya and Arik Bergman
Biosensors 2025, 15(5), 292; https://doi.org/10.3390/bios15050292 - 5 May 2025
Cited by 62 | Viewed by 5381
Abstract
This investigation presents an overview of various optical biosensors utilized for the detection of cancer cells. It covers a comprehensive range of technologies, including surface plasmon resonance (SPR) sensors, which exploit changes in refractive index (RI) [...] Read more.
This investigation presents an overview of various optical biosensors utilized for the detection of cancer cells. It covers a comprehensive range of technologies, including surface plasmon resonance (SPR) sensors, which exploit changes in refractive index (RI) at the sensor surface to detect biomolecular interactions. Localized surface plasmon resonance (LSPR) sensors offer high sensitivity and versatility in detecting cancer biomarkers. Colorimetric sensors, based on color changes induced via specific biochemical reactions, provide a cost-effective and simple approach to cancer detection. Sensors based on fluorescence work using the light emitted from fluorescent molecules detect cancer-specific targets with specificity and high sensitivity. Photonics and waveguide sensors utilize optical waveguides to detect changes in light propagation, offering real-time and label-free detection of cancer biomarkers. Raman spectroscopy-based sensors utilize surface-enhanced Raman scattering (SERS) to provide molecular fingerprint information for cancer diagnosis. Lastly, fiber optic sensors offer flexibility and miniaturization, making them suitable for in vivo and point-of-care applications in cancer detection. This study provides insights into the principles, applications, and advancements of these optical biosensors in cancer diagnostics, highlighting their potential in improving early detection and patient outcomes. Full article
(This article belongs to the Special Issue Fiber Optic Biosensors: Advancements and Applications)
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31 pages, 4112 KB  
Review
Biosensors for Early Detection of Parkinson’s Disease: Principles, Applications, and Future Prospects
by Panpan Jiang, Nan Gao, Gang Chang and Yuxiang Wu
Biosensors 2025, 15(5), 280; https://doi.org/10.3390/bios15050280 - 29 Apr 2025
Cited by 10 | Viewed by 5398
Abstract
Parkinson’s disease (PD), a neurodegenerative disorder marked by the progressive loss of dopaminergic neurons in the substantia nigra, imposes substantial economic burdens, including both direct and indirect costs. The medical community currently lacks a definitive cure for Parkinson’s disease, and early detection is [...] Read more.
Parkinson’s disease (PD), a neurodegenerative disorder marked by the progressive loss of dopaminergic neurons in the substantia nigra, imposes substantial economic burdens, including both direct and indirect costs. The medical community currently lacks a definitive cure for Parkinson’s disease, and early detection is crucial for timely intervention and disease management. As innovative diagnostic tools, biosensors have shown great potential in detecting PD at its early stages. This review comprehensively summarizes recent advances in biosensors for the early detection of PD, with a particular focus on the detection of two key biomarkers: dopamine (DA) and α-synuclein (α-syn). Furthermore, it illustrates a variety of nanotechnology-based biosensors, including optical, electrochemical, and transistor biosensors, detailing their underlying principles, advantages, limitations, and applications in PD detection. Moreover, the review explores the challenges and prospects of advancing biosensors for early PD diagnosis. Full article
(This article belongs to the Section Biosensors and Healthcare)
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24 pages, 1661 KB  
Review
Innovative Methodologies for the Early Detection of Breast Cancer: A Review Categorized by Target Biological Samples
by Antonella Grasso, Vittorio Altomare, Giulia Fiorini, Alessandro Zompanti, Giorgio Pennazza and Marco Santonico
Biosensors 2025, 15(4), 257; https://doi.org/10.3390/bios15040257 - 17 Apr 2025
Cited by 18 | Viewed by 6850
Abstract
Innovative biosensor technologies are revolutionizing cancer detection by offering non-invasive, sensitive, and rapid diagnostic tools, addressing the limitations of conventional screening. Non-invasive samples like breath, saliva, urine, and sweat, analyzed using advanced technologies like electronic nose systems and AI, show promise for early [...] Read more.
Innovative biosensor technologies are revolutionizing cancer detection by offering non-invasive, sensitive, and rapid diagnostic tools, addressing the limitations of conventional screening. Non-invasive samples like breath, saliva, urine, and sweat, analyzed using advanced technologies like electronic nose systems and AI, show promise for early detection and frequent monitoring, though validation is needed. AI integration enhances data analysis and personalization. While blood-based methods remain the gold standard, combining them with less invasive sample types like saliva or sweat, and using sensitive techniques, is a promising direction. Conventional methods (mammography, MRI, etc.) offer proven efficacy, but are costly and invasive. Innovative methods using biosensors offer reduced infrastructure needs, lower costs, and patient-friendly sampling. However, challenges remain in validation, standardization, and low biomarker concentrations. Integrating both methodologies could create a comprehensive framework, combining reliability with accessibility. Future research should focus on robust biosensor development, standardization, expanding application to other cancers, exploring less-studied samples like sweat, and improving affordability for wider adoption, especially in resource-limited settings. The future lies in integrating diverse approaches for more sensitive, specific, and patient-friendly screening, improving early detection and outcomes. Full article
(This article belongs to the Special Issue Innovative Strategies for Cancer Biosensing)
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20 pages, 1451 KB  
Review
Microbial Transcription Factor-Based Biosensors: Innovations from Design to Applications in Synthetic Biology
by Kyeongseok Song, Haekang Ji, Jiwon Lee and Youngdae Yoon
Biosensors 2025, 15(4), 221; https://doi.org/10.3390/bios15040221 - 31 Mar 2025
Cited by 14 | Viewed by 6868
Abstract
Transcription factor-based biosensors (TFBs) are powerful tools in microbial biosensor applications, enabling dynamic control of metabolic pathways, real-time monitoring of intracellular metabolites, and high-throughput screening (HTS) for strain engineering. These systems use transcription factors (TFs) to convert metabolite concentrations into quantifiable outputs, enabling [...] Read more.
Transcription factor-based biosensors (TFBs) are powerful tools in microbial biosensor applications, enabling dynamic control of metabolic pathways, real-time monitoring of intracellular metabolites, and high-throughput screening (HTS) for strain engineering. These systems use transcription factors (TFs) to convert metabolite concentrations into quantifiable outputs, enabling precise regulation of metabolic fluxes and biosynthetic efficiency in microbial cell factories. Recent advancements in TFB, including improved sensitivity, specificity, and dynamic range, have broadened their applications in synthetic biology and industrial biotechnology. Computational tools such as Cello have further revolutionized TFB design, enabling in silico optimization and construction of complex genetic circuits for integrating multiple signals and achieving precise gene regulation. This review explores innovations in TFB systems for microbial biosensors, their role in metabolic engineering and adaptive evolution, and their future integration with artificial intelligence and advanced screening technologies to overcome critical challenges in synthetic biology and industrial bioproduction. Full article
(This article belongs to the Special Issue Microbial Biosensor: From Design to Applications)
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31 pages, 4513 KB  
Review
Fusing Wearable Biosensors with Artificial Intelligence for Mental Health Monitoring: A Systematic Review
by Ali Kargarandehkordi, Shizhe Li, Kaiying Lin, Kristina T. Phillips, Roberto M. Benzo and Peter Washington
Biosensors 2025, 15(4), 202; https://doi.org/10.3390/bios15040202 - 21 Mar 2025
Cited by 41 | Viewed by 13983
Abstract
The development of digital instruments for mental health monitoring using biosensor data from wearable devices can enable remote, longitudinal, and objective quantitative benchmarks. To survey developments and trends in this field, we conducted a systematic review of artificial intelligence (AI) models using data [...] Read more.
The development of digital instruments for mental health monitoring using biosensor data from wearable devices can enable remote, longitudinal, and objective quantitative benchmarks. To survey developments and trends in this field, we conducted a systematic review of artificial intelligence (AI) models using data from wearable biosensors to predict mental health conditions and symptoms. Following PRISMA guidelines, we identified 48 studies using a variety of wearable and smartphone biosensors including heart rate, heart rate variability (HRV), electrodermal activity/galvanic skin response (EDA/GSR), and digital proxies for biosignals such as accelerometry, location, audio, and usage metadata. We observed several technical and methodological challenges across studies in this field, including lack of ecological validity, data heterogeneity, small sample sizes, and battery drainage issues. We outline several corresponding opportunities for advancement in the field of AI-driven biosensing for mental health. Full article
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16 pages, 4073 KB  
Article
Innovative Molecular Imprinting Sensor for Quick, Non-Invasive Cortisol Monitoring in Fish Welfare
by Hugo G. Santos, Daniela Santos Oliveira and Felismina T. C. Moreira
Biosensors 2025, 15(4), 204; https://doi.org/10.3390/bios15040204 - 21 Mar 2025
Cited by 2 | Viewed by 2832
Abstract
The assessment of fish welfare is crucial to prevent economic losses in aquaculture and ensure reliable results in research. A quick, non-invasive device to measure cortisol levels in fish farm water facilitates welfare evaluation and corrective actions when compromised. To address this need, [...] Read more.
The assessment of fish welfare is crucial to prevent economic losses in aquaculture and ensure reliable results in research. A quick, non-invasive device to measure cortisol levels in fish farm water facilitates welfare evaluation and corrective actions when compromised. To address this need, an innovative sensor was developed using screen-printed carbon electrodes (SPCEs) functionalized with reduced graphene oxide/Prussian blue nanocubes (rGO/PBNCs) for direct selective detection of cortisol. A molecularly imprinted polymer (MIP) was synthesized on rGO/PBNCs/SPCEs by electropolymerization (ELP) of pyrrole in the presence of cortisol. The polymerization solution was prepared by adding cortisol (5 mM) and pyrrole (0.3 M) to a DMF/PBS (1:4) solution (pH 7.4). Following ELP, the electrodes were washed with PBS, and pyrrole overoxidation was used to extract cortisol from the polymer matrix. For comparison purposes, a non-imprinted polymer (NIP) was also fabricated. The electrodes were characterized using scanning electron microscopy (SEM) and Raman spectroscopy to assess their morphological and chemical features. Under optimized conditions, the sensor showed a linear range from 0.1 nM to 0.1 mM in artificial saltwater. This sensor combines simplicity and affordability while providing reliable detection of chemical and biological compounds. Full article
(This article belongs to the Special Issue Biosensing Strategies for Food Safety Applications)
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15 pages, 3686 KB  
Article
A Wearable Molecularly Imprinted Electrochemical Sensor for Cortisol Stable Monitoring in Sweat
by Yitao Chen, Zidong He, Yuanzhao Wu, Xinyu Bai, Yuancheng Li, Weiwei Yang, Yiwei Liu and Run-Wei Li
Biosensors 2025, 15(3), 194; https://doi.org/10.3390/bios15030194 - 18 Mar 2025
Cited by 41 | Viewed by 14264
Abstract
Cortisol, a steroid hormone, is closely associated with human mental stress. The rapid, real-time, and continuous detection of cortisol using wearable devices offers a promising approach for individual mental health. These devices must exhibit high sensitivity and long-term stability to ensure reliable performance. [...] Read more.
Cortisol, a steroid hormone, is closely associated with human mental stress. The rapid, real-time, and continuous detection of cortisol using wearable devices offers a promising approach for individual mental health. These devices must exhibit high sensitivity and long-term stability to ensure reliable performance. This study developed a wearable electrochemical sensor based on molecularly imprinted polymer (MIP) technology for real-time and dynamic monitoring of cortisol in sweat. A flexible gold (Au) electrode with interfacial hydrophilic treatment was employed to construct a highly stable electrode. The integration of a silk fibroin/polyvinylidene fluoride (SF/PVDF) composite membrane facilitates directional sweat transport, while liquid metal bonding enhances electrode flexibility and mechanical anti-delamination capability. The sensor exhibits an ultrawide detection range (0.1 pM to 5 μM), high selectivity (over 100-fold against interferents such as glucose and lactic acid), and long-term stability (less than 3.76% signal attenuation over 120 cycles). Additionally, a gradient modulus design was implemented to mitigate mechanical deformation interference under wearable conditions. As a flexible wearable device for cortisol monitoring in human sweat, the sensor’s response closely aligns with the diurnal cortisol rhythm, offering a highly sensitive and interference-resistant wearable solution for mental health monitoring and advancing personalized dynamic assessment of stress-related disorders. Full article
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