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One-Pot Colorimetric Nucleic Acid Test Mediated by Silver Nanoparticles for DNA Extraction and Detection
Rapid and Highly Sensitive Detection of Ricin in Biological Fluids Using Optical Modulation Biosensing
Understanding the Mechanism of Bent DNA Amplifying Sensors Using All-Atom Molecular Dynamics Simulations

Journal Description

Biosensors

Biosensors is an international, peer-reviewed, open access journal on the technology and science of biosensors published monthly online by MDPI.

Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
High Visibility: indexed within Scopus, SCIE (Web of Science), PubMed, MEDLINE, PMC, Embase, CAPlus / SciFinder, Inspec, and other databases.
Journal Rank: JCR - Q1 (Instruments and Instrumentation) / CiteScore - Q1 (Instrumentation)
Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 21.8 days after submission; acceptance to publication is undertaken in 2.8 days (median values for papers published in this journal in the first half of 2025).
Recognition of Reviewers: reviewers who provide timely, thorough peer-review reports receive vouchers entitling them to a discount on the APC of their next publication in any MDPI journal, in appreciation of the work done.

Impact Factor: 5.6 (2024); 5-Year Impact Factor: 5.7 (2024)

Imprint Information Journal Flyer Open Access ISSN: 2079-6374

Latest Articles

40 pages, 3625 KB

Open AccessReview

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 (registering DOI) - 6 Sep 2025

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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26 pages, 6690 KB

Open AccessArticle

Head-Specific Spatial Spectra of Electroencephalography Explained: A Sphara and BEM Investigation

by Uwe Graichen, Sascha Klee, Patrique Fiedler, Lydia Hofmann and Jens Haueisen

Biosensors 2025, 15(9), 585; https://doi.org/10.3390/bios15090585 (registering DOI) - 6 Sep 2025

Electroencephalography (EEG) is a non-invasive biosensing platform with a spatial-frequency content that is of significant relevance for a multitude of aspects in the neurosciences, ranging from optimal spatial sampling of the EEG to the design of spatial filters and source reconstruction. In the [...] Read more.

Electroencephalography (EEG) is a non-invasive biosensing platform with a spatial-frequency content that is of significant relevance for a multitude of aspects in the neurosciences, ranging from optimal spatial sampling of the EEG to the design of spatial filters and source reconstruction. In the past, simplified spherical head models had to be used for this analysis. We propose a method for spatial frequency analysis in EEG for realistically shaped volume conductors, and we exemplify our method with a five-compartment Boundary Element Method (BEM) model of the head. We employ the recently developed technique for spatial harmonic analysis (Sphara), which allows for spatial Fourier analysis on arbitrarily shaped surfaces in space. We first validate and compare Sphara with the established method for spatial Fourier analysis on spherical surfaces, discrete spherical harmonics, using a spherical volume conductor. We provide uncertainty limits for Sphara. We derive relationships between the signal-to-noise ratio (SNR) and the required spatial sampling of the EEG. Our results demonstrate that conventional 10–20 sampling might misestimate EEG power by up to 50%, and even 64 electrodes might misestimate EEG power by up to 15%. Our results also provide insights into the targeting problem of transcranial electric stimulation. Full article

(This article belongs to the Special Issue Biomarkers of Disability and Movement Disorders: Insights from Wearable Devices)

30 pages, 6580 KB

Open AccessArticle

Advanced Nanomaterial-Based Electrochemical Biosensing of Loop-Mediated Isothermal Amplification Products

by Ana Kuprešanin, Marija Pavlović, Ljiljana Šašić Zorić, Milinko Perić, Stefan Jarić, Teodora Knežić, Ljiljana Janjušević, Zorica Novaković, Marko Radović, Mila Djisalov, Nikola Kanas, Jovana Paskaš and Zoran Pavlović

Biosensors 2025, 15(9), 584; https://doi.org/10.3390/bios15090584 (registering DOI) - 5 Sep 2025

The rapid and sensitive detection of regulatory elements within transgenic constructs of genetically modified organisms (GMOs) is essential for effective monitoring and control of their distribution. In this study, we present several innovative electrochemical biosensing platforms for the detection of regulatory sequences in [...] Read more.

The rapid and sensitive detection of regulatory elements within transgenic constructs of genetically modified organisms (GMOs) is essential for effective monitoring and control of their distribution. In this study, we present several innovative electrochemical biosensing platforms for the detection of regulatory sequences in genetically modified (GM) plants, combining the loop-mediated isothermal amplification (LAMP) method with electrodes functionalized by two-dimensional (2D) nanomaterials. The sensor design exploits the high surface area and excellent conductivity of reduced graphene oxide, Ti₃C₂T_x, and molybdenum disulfide (MoS₂) to enhance signal transduction. Furthermore, we used a “green synthesis” method for Ti₃C₂T_x preparation that eliminates the use of hazardous hydrofluoric acid (HF) and hydrochloric acid (HCl), providing a safer and more sustainable approach for nanomaterial production. Within this framework, the performance of various custom-fabricated electrodes, including laser-patterned gold leaf films, physical vapor deposition (PVD)-deposited gold electrodes, and screen-printed gold electrodes, is evaluated and compared with commercial screen-printed gold electrodes. Additionally, gold and carbon electrodes were electrochemically covered by gold nanoparticles (AuNPs), and their properties were compared. Several electrochemical methods were used during the DNA detection, and their importance and differences in excitation signal were highlighted. Electrochemical properties, sensitivity, selectivity, and reproducibility are characterized for each electrode type to assess the influence of fabrication methods and material composition on sensor performance. The developed biosensing systems exhibit high sensitivity, specificity, and rapid response, highlighting their potential as practical tools for on-site GMO screening and regulatory compliance monitoring. This work advances electrochemical nucleic acid detection by integrating environmentally-friendly nanomaterial synthesis with robust biosensing technology. Full article

(This article belongs to the Section Biosensor Materials)

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14 pages, 1621 KB

Open AccessArticle

A Bluetooth-Enabled Electrochemical Platform Based on Saccharomyces cerevisiae Yeast Cells for Copper Detection

by Ehtisham Wahid, Ohiemi Benjamin Ocheja, Antonello Longo, Enrico Marsili, Massimo Trotta, Matteo Grattieri, Cataldo Guaragnella and Nicoletta Guaragnella

Biosensors 2025, 15(9), 583; https://doi.org/10.3390/bios15090583 (registering DOI) - 5 Sep 2025

Copper contamination in the environment poses significant risks to both soil and human health, making the need for reliable monitoring methods crucial. In this study, we report the use of the EmStat Pico module as potentiostat to develop a portable electrochemical biosensor for [...] Read more.

Copper contamination in the environment poses significant risks to both soil and human health, making the need for reliable monitoring methods crucial. In this study, we report the use of the EmStat Pico module as potentiostat to develop a portable electrochemical biosensor for copper detection, utilizing yeast Saccharomyces cerevisiae cells immobilized on a polydopamine (PDA)-coated screen-printed electrode (SPE). By optimizing the sensor design with a horizontal assembly and the volume reduction in the electrolyte solution, we achieved a 10-fold increase in current density with higher range of copper concentrations (0–300 µM CuSO₄) compared to traditional (or previous) vertical dipping setups. Additionally, the use of genetically engineered copper-responsive yeast cells further improved sensor performance, with the recombinant strain showing a 1.7-fold increase in current density over the wild-type strain. The biosensor demonstrated excellent reproducibility (R² > 0.95) and linearity over a broad range of copper concentrations, making it suitable for precise quantitative analysis. To further enhance portability and usability, a Bluetooth-enabled electrochemical platform was integrated with a web application for real-time data analysis, enabling on-site monitoring and providing a reliable, cost-effective tool for copper detection in real world settings. This system offers a promising solution for addressing the growing need for efficient environmental monitoring, especially in agriculture. Full article

(This article belongs to the Special Issue Sensors for Environmental Monitoring and Food Safety—2nd Edition)

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14 pages, 3684 KB

Open AccessArticle

Accuracy Enhancement in Refractive Index Sensing via Full-Spectrum Machine Learning Modeling

by Majid Aalizadeh, Chinmay Raut, Morteza Azmoudeh Afshar, Ali Tabartehfarahani and Xudong Fan

Biosensors 2025, 15(9), 582; https://doi.org/10.3390/bios15090582 - 5 Sep 2025

We present a full-spectrum machine learning framework for refractive index sensing using simulated absorption spectra from meta-grating structures composed of titanium or silicon nanorods under TE and TM polarizations. Linear regression was applied to 80 principal components extracted from each spectrum, and model [...] Read more.

We present a full-spectrum machine learning framework for refractive index sensing using simulated absorption spectra from meta-grating structures composed of titanium or silicon nanorods under TE and TM polarizations. Linear regression was applied to 80 principal components extracted from each spectrum, and model performance was assessed using five-fold cross-validation, simulating real-world biosensing scenarios where unknown patient samples are predicted based on standard calibration data. Titanium-based structures, dominated by broadband intensity changes, yielded the lowest mean squared errors and the highest accuracy improvements—up to an 8128-fold reduction compared to the best single-feature model. In contrast, silicon-based structures, governed by narrow resonances, showed more modest gains due to spectral nonlinearity that limits the effectiveness of global linear models. We also show that even the best single-wavelength predictor is identified through data-driven analysis, not visual selection, highlighting the value of automated feature preselection. These findings demonstrate that spectral shape plays a key role in modeling performance and that full-spectrum linear approaches are especially effective for intensity-modulated index sensors. Full article

(This article belongs to the Section Optical and Photonic Biosensors)

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16 pages, 3627 KB

Open AccessArticle

In Vivo Study on the Safe Use of a Novel Intraoperative Sensing Tool for Tissue Stiffness Assessment in Endoscopic Surgery

by Georgios Violakis, Pantelis Antonakis, Emmanouil Kritsotakis, Theodoros Kozonis, Leonidas Chardalias, Apostolos Papalois, Georgios Agrogiannis, Effrosyni Kampouroglou, Nikolaos Vardakis, Stylianos Kostakis, Eleni Athanasaki, Zhenyu Zhang, Martin Angelmahr, Manousos Konstadoulakis and Panagiotis Polygerinos

Biosensors 2025, 15(9), 581; https://doi.org/10.3390/bios15090581 - 5 Sep 2025

A novel endoscopic palpation tool (EPT), designed for tactile and stiffness sensing using fiber Bragg gratings (FBGs) was evaluated in a surgical environment for intraoperative safety and effectiveness. The EPT consisted of four FBGs arranged in a cross pattern and embedded within an [...] Read more.

A novel endoscopic palpation tool (EPT), designed for tactile and stiffness sensing using fiber Bragg gratings (FBGs) was evaluated in a surgical environment for intraoperative safety and effectiveness. The EPT consisted of four FBGs arranged in a cross pattern and embedded within an elastic, hollow, silicone hemispherical dome designed to deform upon contact with soft tissue. The EPT was employed to scan both in vivo and ex vivo tissue samples. Monitoring of porcine vital signs during minimally invasive and open surgical procedures showed no significant changes during use of the EPT. Perioperative blood tests including inflammatory markers and liver and renal function studies were unremarkable. Histopathological analyses of tissues involved (liver, spleen, bowel, and abdominal wall) showed no evidence of inflammation, necrosis, or tissue damage, confirming the device’s biocompatibility. To the best of our knowledge, this is the first study reporting in vivo stiffness measurements using an FBG-based EPT. The probe successfully distinguished between soft and hard tissue regions’ relative stiffness. Furthermore, successive measurements on liver samples demonstrated the device’s ability to generate stiffness maps, enabling clear visualization of spatial variation in tissue stiffness. Full article

(This article belongs to the Section Optical and Photonic Biosensors)

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32 pages, 5916 KB

Open AccessReview

Advances and Innovations in Conjugated Polymer Fluorescent Sensors for Environmental and Biological Detection

by Viet-Duc Phung and Vinh Van Tran

Biosensors 2025, 15(9), 580; https://doi.org/10.3390/bios15090580 - 4 Sep 2025

Thanks to their multiple outstanding features—such as high fluorescence quantum yield, good photostability, and excellent sensitivity—conjugated polymers (CPs) have emerged as a pioneering class of fluorescent materials for sensing applications, particularly in environmental and biological fields, for the detection of a wide range [...] Read more.

Thanks to their multiple outstanding features—such as high fluorescence quantum yield, good photostability, and excellent sensitivity—conjugated polymers (CPs) have emerged as a pioneering class of fluorescent materials for sensing applications, particularly in environmental and biological fields, for the detection of a wide range of environmental pollutants and bioactive compounds. The presence of delocalized π-electrons in the CP backbone significantly enhances sensing performance through a unique phenomenon known as the “molecular wire effect.” As a result, CP-based fluorescent sensors have been extensively developed and employed as exceptional tools for monitoring various analytes in environmental and biological contexts. A deep understanding of their unique properties, fabrication techniques, and recent innovations is essential for guiding the strategic development of advanced CP-based fluorescent sensors, particularly for future point-of-care applications. This study presents a critical review of the key characteristics of fluorescent sensors and highlights several common types of conjugated polymers (CPs) used in their design and fabrication. It summarizes and discusses the main sensing mechanisms, state-of-the-art applications, and recent innovations of CP-based fluorescent sensors for detecting target compounds in environmental and biological fields. Furthermore, potential strategies and future perspectives for designing and developing high-performance CP-based fluorescent sensors are emphasized. By consolidating current scientific evidence, this review aims to support the advancement of highly sensitive fluorescent sensors based on various CP nanoparticles for environmental and biological applications. Full article

(This article belongs to the Special Issue Polymers-Based Biosensors and Bioelectronics: Designs and Applications)

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17 pages, 1568 KB

Open AccessArticle

Cell-Free DNA Versus Circulating Tumor Cells: A Pilot Study of Alpha-Fetoprotein Analysis for Diagnosis and Treatment Monitoring in Hepatocellular Carcinoma

by Ga Young Moon, Hyun Sung Park, Ha Neul Kim, Hei-Gwon Choi, Yonghan Han, Hyuk Soo Eun, Tae Hee Lee and Jiyoon Bu

Biosensors 2025, 15(9), 579; https://doi.org/10.3390/bios15090579 - 4 Sep 2025

Serum alpha-fetoprotein (AFP) is widely used for hepatocellular carcinoma (HCC) management, yet its limited sensitivity and specificity restrict diagnostic and prognostic utility. In this study, we explore the clinical potential of AFP quantification from cell-free DNA (cfDNA) and circulating tumor cells (CTCs) using [...] Read more.

Serum alpha-fetoprotein (AFP) is widely used for hepatocellular carcinoma (HCC) management, yet its limited sensitivity and specificity restrict diagnostic and prognostic utility. In this study, we explore the clinical potential of AFP quantification from cell-free DNA (cfDNA) and circulating tumor cells (CTCs) using a novel bead-based liquid biopsy platform. Following isolation, AFP abundance in cfDNA was quantified by qPCR, while AFP protein expression in CTCs was assessed via immunohistochemistry. Compared to serum AFP, cfDNA-derived AFP demonstrated significantly greater diagnostic accuracy in distinguishing HCC patients from non-cancerous individuals (p < 0.0001, AUC = 0.998), while AFP⁺ CTCs showed high specificity. Post-treatment changes in AFP levels from cfDNA and CTCs were significantly associated with therapeutic response and overall survival, outperforming conventional serum AFP. Longitudinal monitoring further revealed that cfDNA AFP levels reliably captured recurrence events prior to clinical diagnosis. Moreover, a combined metric integrating AFP levels from cfDNA and CTCs significantly improved response stratification (AUC = 0.89), outperforming individual biomarkers. This pilot study highlights the potential of multimodal AFP profiling through cfDNA and CTCs as a promising, non-invasive approach for enhancing diagnosis, prognosis, and treatment monitoring in HCC, with direct implications for personalized therapeutic strategies. Full article

(This article belongs to the Special Issue Advanced Cell-Analyzing Technologies and Their Biosensing Applications)

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Graphical abstract

22 pages, 4751 KB

Open AccessArticle

Motion Artifacts (MA) At-Rest in Measured Arterial Pulse Signals: Time-Varying Amplitude in Each Harmonic and Non-Flat Harmonic-MA-Coupled Baseline

by MD Mahfuzur Rahman, Mamun Hasan and Zhili Hao

Biosensors 2025, 15(9), 578; https://doi.org/10.3390/bios15090578 - 4 Sep 2025

Motion artifacts (MA) cause great variability in a measured arterial pulse signal, and treatment of MA solely as a baseline drift (BD) fails to eliminate its effect on the measured signal. This paper presents a study on the effect of MA at rest [...] Read more.

Motion artifacts (MA) cause great variability in a measured arterial pulse signal, and treatment of MA solely as a baseline drift (BD) fails to eliminate its effect on the measured signal. This paper presents a study on the effect of MA at rest (<0.7 Hz) on measured arterial pulse signals using a microfluidic-based tactile sensor. By taking full account of the dynamic behavior of the transmission path from the true pulse signal in an artery to a measured pulse signal at the sensor, the tissue-contact-sensor (TCS) stack, an analytical model of MA in a measured pulse signal is developed. In this model, the TCS stack is treated as a 1DOF system for its dynamic behavior; MA is quantified as the displacement (i.e., BD) and time-varying system parameters (TVSP) of the TCS stack. The mathematical expression of MA in a measured pulse signal reveals that while BD remains as low-frequency additive noise, TVSP causes time-varying harmonics in a measured pulse signal. Further time-frequency analysis (TFA) of measured pulse signals validates the existence of TVSP and, for the first time, reveals its effect on a measured pulse signal: time-varying amplitude in each harmonic and non-flat harmonic-MA-coupled baseline. Full article

(This article belongs to the Special Issue Biosensors Based on Microfluidic Devices—2nd Edition)

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17 pages, 1545 KB

Open AccessArticle

Portable Point-of-Care Device for Dual Detection of Glucose-6-Phosphate Dehydrogenase Deficiency and Hemoglobin in Low-Resource Settings

by Rehab Osman Taha, Napaporn Youngvises, Runtikan Pochairach, Papichaya Phompradit and Kesara Na-Bangchang

Biosensors 2025, 15(9), 577; https://doi.org/10.3390/bios15090577 - 3 Sep 2025

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common enzymopathy with significant clinical implications, particularly in malaria-endemic regions and in the management of neonatal hyperbilirubinemia. Timely and accurate detection of G6PD deficiency is critical to prevent life-threatening hemolytic events following oxidative drug administration. This study [...] Read more.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common enzymopathy with significant clinical implications, particularly in malaria-endemic regions and in the management of neonatal hyperbilirubinemia. Timely and accurate detection of G6PD deficiency is critical to prevent life-threatening hemolytic events following oxidative drug administration. This study evaluated the MyG6PD device, a quantitative point-of-care (PoC) tool, for the assessment of hemoglobin concentration and G6PD enzyme activity. Analytical performance was benchmarked against laboratory spectrophotometry and the STANDARD G6PD Analyzer™ (SD Biosensor; Suwon-si, Republic of Korea). MyG6PD demonstrated excellent linearity (R² ≥ 0.99), accuracy (bias < ±15%), and precision (CV < 15%) across normal, intermediate, and deficient activity ranges, including heterozygous females with intermediate phenotypes. The device’s compact, battery-operated design, rapid turnaround, and minimal training requirements support its use in decentralized and resource-limited settings. Furthermore, cost-effective consumables and robust detection of intermediate activity highlight its potential for large-scale deployment. Overall, MyG6PD provides a reliable, accessible, and clinically actionable solution for urgent G6PD deficiency screening, enabling safer administration of oxidative therapies and improving patient outcomes in high-risk populations. Full article

(This article belongs to the Section Biosensors and Healthcare)

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17 pages, 2747 KB

Open AccessArticle

Flexible and Stretchable Microneedle Electrode Arrays by Soft Lithography for Continuous Monitoring of Glucose

by Yong-Ho Choi, Honglin Piao, Jia Lee, Jaehyun Kim, Heon-Jin Choi and Dahl-Young Khang

Biosensors 2025, 15(9), 576; https://doi.org/10.3390/bios15090576 - 2 Sep 2025

Continuous monitoring of glucose (CGM) level is of utmost importance to diabetic patients, especially with no or minimal pain. Microneedle arrays with desired electrode patterns have been fabricated by soft lithographic molding, and the patterned electrodes were formed via shadow evaporation through a [...] Read more.

Continuous monitoring of glucose (CGM) level is of utmost importance to diabetic patients, especially with no or minimal pain. Microneedle arrays with desired electrode patterns have been fabricated by soft lithographic molding, and the patterned electrodes were formed via shadow evaporation through a shadow mask that was made from a modified molding technique. With immobilization of glucose oxidase (GOx), the microneedle electrode arrays (MEAs) have been successfully employed for the in vitro CGM using impedance spectroscopy. The fabricated MEAs could monitor the varying glucose level continuously for up to ~10 days. Similar processes have been applied for the fabrication of stretchable MEAs, which can conform to complex curvilinear surfaces. The simple and low-cost fabrication of MEAs, either in flexible or stretchable forms, may find various applications in wearable health monitoring techniques. Full article

(This article belongs to the Special Issue Recent Advances in Glucose Biosensors)

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22 pages, 2596 KB

Open AccessArticle

A Sensitive Electrochemical Cholinesterase-Inhibiting Biosensor for Organophosphorus Pesticides Based on Ti₃C₂T_X 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

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 Ti₃C₂T_x 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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33 pages, 4561 KB

Open AccessReview

Smartphone-Integrated Electrochemical Devices for Contaminant Monitoring in Agriculture and Food: A Review

by Sumeyra Savas and Seyed Mohammad Taghi Gharibzahedi

Biosensors 2025, 15(9), 574; https://doi.org/10.3390/bios15090574 - 2 Sep 2025

Recent progress in microfluidic technologies has led to the development of compact and highly efficient electrochemical platforms, including lab-on-a-chip (LoC) systems, that integrate multiple testing functions into a single, portable device. Combined with smartphone-based electrochemical devices, these systems enable rapid and accurate on-site [...] Read more.

Recent progress in microfluidic technologies has led to the development of compact and highly efficient electrochemical platforms, including lab-on-a-chip (LoC) systems, that integrate multiple testing functions into a single, portable device. Combined with smartphone-based electrochemical devices, these systems enable rapid and accurate on-site detection of food contaminants, including pesticides, heavy metals, pathogens, and chemical additives at farms, markets, and processing facilities, significantly reducing the need for traditional laboratories. Smartphones improve the performance of these platforms by providing computational power, wireless connectivity, and high-resolution imaging, making them ideal for in-field food safety testing with minimal sample and reagent requirements. At the core of these systems are electrochemical biosensors, which convert specific biochemical reactions into electrical signals, ensuring highly sensitive and selective detection. Advanced nanomaterials and integration with Internet of Things (IoT) technologies have further improved performance, delivering cost-effective, user-friendly food monitoring solutions that meet regulatory safety and quality standards. Analytical techniques such as voltammetry, amperometry, and impedance spectroscopy increase accuracy even in complex food samples. Moreover, low-cost engineering, artificial intelligence (AI), and nanotechnology enhance the sensitivity, affordability, and data analysis capabilities of smartphone-integrated electrochemical devices, facilitating their deployment for on-site monitoring of food and agricultural contaminants. This review explains how these technologies address global food safety challenges through rapid, reliable, and portable detection, supporting food quality, sustainability, and public health. Full article

(This article belongs to the Special Issue Electrochemical (Bio)Sensors as Promising Analytical Tools in the Analysis of Soils, Plants and Environmental Monitoring)

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23 pages, 3035 KB

Open AccessReview

SERS-Driven Evolution of Lateral and Vertical Flow Assays in Medical Diagnostics

by Boyou Heo and Ho Sang Jung

Biosensors 2025, 15(9), 573; https://doi.org/10.3390/bios15090573 - 1 Sep 2025

Surface-enhanced Raman scattering (SERS) has emerged as a powerful signal amplification strategy to address the inherent limitations of conventional flow-based diagnostic methods such as lateral flow analysis (LFA) and vertical flow analysis (VFA). By incorporating SERS-active nanostructures into these platforms, SERS-integrated LFA and [...] Read more.

Surface-enhanced Raman scattering (SERS) has emerged as a powerful signal amplification strategy to address the inherent limitations of conventional flow-based diagnostic methods such as lateral flow analysis (LFA) and vertical flow analysis (VFA). By incorporating SERS-active nanostructures into these platforms, SERS-integrated LFA and VFA systems have significantly improved sensitivity, specificity, and multiplexing performance while maintaining the simplicity and portability of conventional approaches. In this review, we summarize recent advances in SERS-enhanced flow-based diagnostics with a focus on exogenous and endogenous disease detection. Exogenous targets include viral antigens, bacterial pathogens, and foodborne contaminants such as mycotoxins and antibiotic residues. Endogenous applications include therapeutic drug monitoring, inflammation profiling, cancer biomarker detection, and exosome-based molecular subtyping. We highlight the structural differences between LFA and VFA approaches and their impact on analytical performance, and explore the advantages of SERS-integrated platforms for rapid and multiplexed detection in complex biological matrices. Finally, we provide an overview of key technical challenges, such as signal reproducibility, matrix interference, and device integration, and discuss future directions for clinical implementation of SERS-based flow diagnostics in point-of-care settings. Full article

(This article belongs to the Special Issue Nano/Micro Biosensors for Biomedical Applications (2nd Edition))

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13 pages, 2327 KB

Open AccessCommunication

Quantitative IC₅₀ Analysis of Puromycin-Induced Cytotoxicity in NIH/3T3 Cells Using a Multi-Well Array Impedance Biosensor

by Seok-kyu Kim, SuGwon Nam and Moongyu Jang

Biosensors 2025, 15(9), 572; https://doi.org/10.3390/bios15090572 - 1 Sep 2025

ECIS-based impedance biosensors have been extensively studied in various fields including cancer research, microbiology, and immunology. However, most studies have primarily focused on monitoring cellular behavior through impedance changes, with relatively less emphasis on interpreting the biological significance of impedance signals. In this [...] Read more.

ECIS-based impedance biosensors have been extensively studied in various fields including cancer research, microbiology, and immunology. However, most studies have primarily focused on monitoring cellular behavior through impedance changes, with relatively less emphasis on interpreting the biological significance of impedance signals. In this study, we employed a multi-well array impedance biosensor to conduct IC₅₀ (half-maximal inhibitory concentration) analysis, a widely used metric for evaluating drug efficacy and toxicity in biological and pharmacological research. Specifically, we assessed the IC₅₀ values of puromycin, an aminonucleoside antibiotic known to inhibit protein synthesis. NIH/3T3 fibroblasts were exposed to various concentrations of puromycin, and real-time impedance monitoring was performed. Cell viability was assessed, and the IC₅₀ value of puromycin for NIH/3T3 cells was determined to be 3.96 µM using capacitance-based impedance analysis. Our findings demonstrate that the multi-well array impedance biosensor provides a rapid and quantitative method for drug toxicity evaluation, offering a valuable platform for drug screening and biocompatibility assessment. Full article

(This article belongs to the Special Issue Cutting-Edge Nanozyme Biosensing Strategies for Biomedical and Environmental Applications)

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29 pages, 3451 KB

Open AccessReview

Deep Learning-Enhanced Nanozyme-Based Biosensors for Next-Generation Medical Diagnostics

by Seungah Lee, Nayra A. M. Moussa and Seong Ho Kang

Biosensors 2025, 15(9), 571; https://doi.org/10.3390/bios15090571 - 1 Sep 2025

The integration of deep learning (DL) and nanozyme-based biosensing has emerged as a transformative strategy for next-generation medical diagnostics. This review explores how DL architectures enhance nanozyme design, functional optimization, and predictive modeling by elucidating catalytic mechanisms such as dual-atom active sites and [...] Read more.

The integration of deep learning (DL) and nanozyme-based biosensing has emerged as a transformative strategy for next-generation medical diagnostics. This review explores how DL architectures enhance nanozyme design, functional optimization, and predictive modeling by elucidating catalytic mechanisms such as dual-atom active sites and substrate-surface interactions. Key applications include disease biomarker detection, medical imaging enhancement, and point-of-care diagnostics aligned with the ASSURED criteria. In clinical contexts, advances such as wearable biosensors and smart diagnostic platforms leverage DL for real-time signal processing, pattern recognition, and adaptive decision-making. Despite significant progress, challenges remain—particularly the need for standardized biomedical datasets, improved model robustness across diverse populations, and the clinical translation of artificial intelligence (AI)-enhanced nanozyme systems. Future directions include integration with the Internet of Medical Things, personalized medicine frameworks, and sustainable sensor development. The convergence of nanozymes and DL offers unprecedented opportunities to advance intelligent biosensing and reshape precision diagnostics in healthcare. Full article

(This article belongs to the Special Issue Cutting-Edge Nanozyme Biosensing Strategies for Biomedical and Environmental Applications)

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19 pages, 3335 KB

Open AccessArticle

CH₃COOAg with Laccase-like Activity for Differentiation and Detection of Aminoglycoside Antibiotics

by Huan Zhu, Tong-Qing Chai, Jia-Xin Li, Jing-Jing Dai, Lei Xu, Wen-Ling Qin and Feng-Qing Yang

Biosensors 2025, 15(9), 570; https://doi.org/10.3390/bios15090570 - 1 Sep 2025

Aminoglycoside antibiotics (AGs) are widely used in medicine and animal husbandry, but they pose significant risks due to residual toxicity and antibiotic resistance. In this study, a novel chemical sensor based on the laccase-like activity of CH₃COOAg was developed for the [...] Read more.

Aminoglycoside antibiotics (AGs) are widely used in medicine and animal husbandry, but they pose significant risks due to residual toxicity and antibiotic resistance. In this study, a novel chemical sensor based on the laccase-like activity of CH₃COOAg was developed for the selective detection of AGs. CH₃COOAg exhibited varying degrees of laccase-like activity in different buffers (MES, HEPES, and NaAc) and H₂O, and five AGs showed distinct intensities of the inhibitory effect on the laccase-like activity of CH₃COOA in different buffers and H₂O. Therefore, a four-channel colorimetric sensor array was constructed in combination with the use of principal component analysis (PCA) and Hierarchical Cluster Analysis (HCA) for the efficient identification of five AGs (0.02–0.3 μM) in environment samples like tap and lake water. In addition, a colorimetric method was developed for kanamycin (KAN) detection in a honey sample with a linear range of 10–100 nM (R² = 0.9977). The method has excellent sensitivity with a limit of detection of 3.99 nM for KAN. This work not only provides a rapid and low-cost detection method for AG monitoring but also provides a reference for the design of non-copper laccase mimics. Full article

(This article belongs to the Special Issue Biosensors for Environmental Monitoring and Food Safety)

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21 pages, 6094 KB

Open AccessArticle

Nanopore-Aware Embedded Detection for Mobile DNA Sequencing: A Viterbi–HMM Design Versus Deep Learning Approaches

by Karim Hammad, Zhongpan Wu, Ebrahim Ghafar-Zadeh and Sebastian Magierowski

Biosensors 2025, 15(9), 569; https://doi.org/10.3390/bios15090569 - 1 Sep 2025

Nanopore-based DNA sequencing has emerged as a transformative biosensing technology, enabling real-time molecular diagnostics in compact and mobile form factors. However, the computational complexity of the basecalling process—the step that translates raw nanopore signals into nucleotide sequences—poses a critical energy challenge for mobile [...] Read more.

Nanopore-based DNA sequencing has emerged as a transformative biosensing technology, enabling real-time molecular diagnostics in compact and mobile form factors. However, the computational complexity of the basecalling process—the step that translates raw nanopore signals into nucleotide sequences—poses a critical energy challenge for mobile deployment. While deep learning (DL) models currently dominate this task due to their high accuracy, they demand substantial power budgets and computing resources, making them unsuitable for portable or field-scale biosensor platforms. In this work, we propose an embedded hardware–software framework for DNA sequence detection that leverages a Viterbi-based Hidden Markov Model (HMM) implemented on a custom 64-bit RISC-V core. The proposed HMM detector is realized on an off-the-shelf Virtex-7 FPGA and evaluated against state-of-the-art DL-based basecallers in terms of energy efficiency and inference accuracy. From one side, the experimental results show that our system achieves an energy efficiency improvement of 6.5×, 5.5×, and 4.6×, respectively, compared to similar HMM-based detectors implemented on a commodity x86 processor, Cortex-A9 ARM embedded system, and a previously published Rocket-based system. From another side, the proposed detector demonstrates 15× and 2.4× energy efficiency superiority over state-of-the-art DL-based detectors, with competitive accuracy and sufficient throughput for field-based genomic surveillance applications and point-of-care diagnostics. This study highlights the practical advantages of classical probabilistic algorithms when tightly integrated with lightweight embedded processors for biosensing applications constrained by energy, size, and latency. Full article

(This article belongs to the Special Issue Novel Nanomaterials and Nanotechnology: From Fabrication Methods and Improvement Strategies to Applications in Biosensing and Biomedicine (2nd Edition))

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20 pages, 10282 KB

Open AccessArticle

A Highly Sensitive SERS Technique Based on Au NPs Monolayer Film Combined with Multivariate Statistical Algorithms for Auxiliary Screening of Postmenopausal Osteoporosis

by Yun Yu, Jinlian Hu, Qidan Shen, Huifeng Xu, Shanshan Wang, Xiaoning Wang, Yuhuan Zhong, Tingting He, Hao Huang, Quanxing Hong, Erdan Huang and Xihai Li

Biosensors 2025, 15(9), 568; https://doi.org/10.3390/bios15090568 - 30 Aug 2025

Postmenopausal osteoporosis (PMOP) has become an important public health issue. The diagnosis of PMOP relies on clinical symptoms and radiology. However, most patients with PMOP do not exhibit obvious symptoms in the early stages of this disease. This study aimed to explore the [...] Read more.

Postmenopausal osteoporosis (PMOP) has become an important public health issue. The diagnosis of PMOP relies on clinical symptoms and radiology. However, most patients with PMOP do not exhibit obvious symptoms in the early stages of this disease. This study aimed to explore the feasibility of surface-enhanced Raman scattering (SERS) technology in the auxiliary screening of PMOP. PMOP rats were induced by ovariectomy (OVX) surgery, with a Sham group and an icariin (ICA) treatment group serving as controls. A monolayer film of Au nanoparticles (NPs) was prepared using the Marangoni effect in an oil/water/oil three-phase system, and was used to detect serum SERS signals in the Sham, OVX, and ICA treatment groups. Then, the spectral diagnostic model for PMOP screening was established utilizing partial least squares (PLS) and support vector machine (SVM) algorithms. Histopathology confirmed the establishment of the PMOP rat model. The assignment of Raman peaks and the analysis of spectral differences revealed the biochemical changes associated with PMOP, including the upregulation of tyrosine levels and the downregulation of arginine, tryptophan, lipids, and collagen. When employing the PLS-SVM algorithm to simultaneously classify and discriminate three groups of samples, the diagnostic sensitivity for PMOP is 93.33%, the specificity is 96.67%, and the accuracy of three-class classification is 91.11%. This study demonstrated the potential of SERS for the auxiliary screening of PMOP. Full article

(This article belongs to the Special Issue Surface-Enhanced Raman Scattering in Biosensing Applications)

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17 pages, 2925 KB

Open AccessArticle

Case Study on Skin Calorimetry: Modeling Localized Muscle Heat Transfer During Exercise

by Pedro Jesús Rodríguez de Rivera, Miriam Rodríguez de Rivera, Fabiola Socorro and Manuel Rodríguez de Rivera

Biosensors 2025, 15(9), 567; https://doi.org/10.3390/bios15090567 - 29 Aug 2025

Direct measurement of heat loss in a moving limb requires attached heat-flux sensors, which are strongly affected by convection and radiation. Skin calorimetry minimizes these effects, enabling an accurate measurement. A skin calorimeter was used to measure the heat flux in the rectus [...] Read more.

Direct measurement of heat loss in a moving limb requires attached heat-flux sensors, which are strongly affected by convection and radiation. Skin calorimetry minimizes these effects, enabling an accurate measurement. A skin calorimeter was used to measure the heat flux in the rectus femoris (thigh) of a subject exercising for 30 min at a mechanical power of 80 W. In this work, we have developed an analytical model able to describe the thermal evolution of the rectus femoris during exercise and subsequent recovery. This model consists of a sum of two exponentials f(t) = A₁(1 − e^−t/τ) + A₂·t·e^−t/τ, with the novelty that the second term is a linear–exponential, which opposes the first term, and that allows the initial thermal transient characterization. The time constants are the most relevant parameters, with mean values of 5 min during exercise and 10 min during recovery (for the 4 cm² sensing area). The mean exercise amplitude (A₁) is 1.1 mW/W, while in post-exercise it is −0.8 mW/W. In addition, the measurement of the thermal resistance of the skin before and after exercise allowed for the estimation and analysis of the evolution of the subcutaneous internal temperature, which follows the same exponential function. The developed mathematical model defines a Transfer Function (TF)—a potential invariant that can predict the thigh’s heat flux response to any exercise protocol (for the subject analyzed). This mathematical approach may be useful for sports and clinical applications. Full article

(This article belongs to the Section Wearable Biosensors)

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