Search Results (308)

As a noninvasive liquid biopsy approach, extracellular vesicle (EV)-based detection offers significant advantages in reflecting real-time tumor dynamis and overcoming the limitations of conventional tissue biopsy. EVs, nanoscale vesicles secreted by cells, carry diverse biomolecules such as proteins and nucleic acids, playing key roles in tumor progression, metastasis, and immune evasion, and have emerged as promising biomarkers for cancer liquid biopsy. Surface-enhanced Raman spectroscopy (SERS), characterized by high sensitivity, resistance to photobleaching, minimal sample consumption, and multiplexing capability, has shown great potential in EV analysis. This review systematically summarizes current methods for EV isolation, characterization, and storage, with a focus on label-free and label-based SERS detection strategies for early cancer diagnosis, treatment response monitoring, and prognosis evaluation. Furthermore, the integration of SERS with machine learning and deep learning algorithms has substantially improved diagnostic accuracy and cancer subtyping. Despite remaining challenges, such as optimization of SERS substrate performance, intelligent processing of Raman spectral fingerprints, and clinical translation, EV-based SERS technology holds great promise for precision oncology. Full article

Localized surface plasmon resonance biosensors are promising devices for label-free detection of live cells and biomolecules. However, typical plasmonic sensors have limited surface area, planar electromagnetic fields, and poor compatibility with three-dimensional (3D) interactions with cells or biomolecules. In this study, a 3D plasmonic sensor around microposts was developed using reversal nanoimprint lithography for highly sensitive cell and DNA detection. Au nanopillars were conformally integrated onto the bottom, sidewall, and top of microposts, forming additional sensing surface area along the sidewall of microposts for plasmonic sensing. The 3D plasmonic sensors exhibited tunable resonance peaks and refractive index (RI) sensitivities by varying the micropost height. The highest sensitivity of 1306 nm per RI unit was obtained from the sensor with 10 μm-tall microposts at a resonance wavelength of 1315 nm, which was significantly higher than that of typical planar plasmonic sensors. The platform was applied to live MC3T3-E1 cell detection, showing a resonance peak shift of 71 ± 11.6 nm at a cell concentration of 10⁶ cells/mL with a cell concentration ranging from 10² to 10⁶ cells/mL. In addition, DNA hybridization detection was demonstrated over a concentration range of 10⁻¹⁵–10⁻⁷ M complementary target DNA, with a resonance shift of 68 ± 2.5 nm observed at 10⁻⁷ M target DNA concentration. The 3D plasmonic sensor provides a scalable device for additional plasmonic biointerfaces with enhanced analyte accessibility and light–matter interactions. This platform offers high-sensitivity biosensing involving live cells, nucleic acids, and other biological targets. Full article

In medicine, nanoparticles are used for various purposes, including theranostics, imaging, diagnostics, drug delivery, tissue regeneration and targeted cancer treatments, and to minimize the harmful side effects associated with conventional therapies. Target-specific biomolecules, such as silica nanoparticles (SiNPs) labeled with metallic radionuclides, are becoming increasingly popular. The choice of radionuclide is based on its nuclear properties. Silica has several advantages for nanoparticle synthesis, including high biocompatibility, the capacity for drug encapsulation due to its porous structure, and the potential for extensive surface functionalization, including radiolabeling for imaging and therapeutic applications. A radionuclide can be attached to a silica nanoparticle either directly or through the use of chelators or polymers. Additionally, the capability to encapsulate therapeutic agents within such systems offers significant potential for the development of targeted therapies. This study aims to provide a comprehensive overview of recent developments in the radiolabeling of silica-based nanoparticles, with a focus on their application in nuclear medicine, particularly in diagnostic imaging and targeted radionuclide therapy. Theranostics employs a range of imaging modalities to guide and monitor therapeutic interventions. Principal techniques include positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), and Optical Imaging (such as fluorescence and bioluminescence). These imaging methods enable precise visualization of pathological sites, facilitate tracking of therapeutic agent distribution, and permit real-time assessment of treatment efficacy. Full article

Fluorine-18 is often considered an ideal positron emitter owing to its excellent chemical, physiological, and nuclear properties. Consequently, the development of rapid, simple, and reliable ¹⁸F-labeling strategies remains critically important for synthesizing new radiopharmaceuticals for PET molecular imaging. A common approach involves the synthesis of ¹⁸F-labeled prosthetic groups that subsequently undergo bioconjugation with peptides or other biomolecules to generate ¹⁸F-labeled imaging probes. However, conventional synthetic methods for these prosthetic groups are often lengthy, require large quantities of precursor and solvent, and typically rely on elevated reaction temperatures. Herein, we report a droplet-based microscale synthetic methodology for the preparation of the [¹⁸F]FVSB prosthetic group that minimizes precursor and solvent usage, proceeds rapidly, and operates at relatively low temperatures. Conditions were optimized using a platform for performing droplet reactions in parallel, enabling high-throughput study of multiple reaction parameters within a short period of time. Additionally, we introduce a simple micro-cartridge purification technique that affords purified [¹⁸F]FVSB in small volumes. Furthermore, we describe an efficient bioconjugation that requires substantially lower reagent amounts than the previously reported macroscale method. The microscale process we report could facilitate wider use of this ¹⁸F-labeling strategy and can be extended to label other thiol-bearing peptides or biomolecules. Full article

The development of reliable electrochemical interfaces for biosensor applications requires materials that combine high conductivity, large effective surface area, and suitable platforms for biomolecule immobilization. In this work, a hybrid electrochemical platform based on screen-printed carbon electrodes (SPCEs) modified with electropolymerized polypyrrole (PPy) and electrodeposited silver particles (AgPs) is presented for the subsequent immobilization of Ki-67 antibodies. PPy films were synthesized under optimized electrochemical conditions, producing homogeneous, porous, and electrochemically stable coatings that significantly enhanced the doping/undoping processes from 0.3280 C/0.3284 C to 0.3281 C/0.3284 C for SPCE and SPCE-PPy, respectively. Subsequently, silver particles were deposited onto the PPy matrix, resulting in a well-dispersed and uniform distribution of AgPs, promoted by the interaction between Ag⁰ and the nitrogen groups in the polymer backbone. The synergistic combination of PPy and AgPs resulted in improved charge-transfer properties and enhanced electrochemical reversibility, thereby decreasing the peak-to-peak separation of the ferricyanide/ferrocyanide redox couple used as a probe by 40%. Immobilization of Ki-67 antibodies was achieved via direct interaction with AgPs, resulting in a marked passivation effect, as evidenced by the suppression of redox probe signals, confirming successful biofunctionalization. The proposed SPCE-PPy-AgP architecture provides a robust, reproducible, and versatile platform for antibody immobilization, as demonstrated by oxidation and reduction peaks with relative standard deviations (RSDs) of 3.18% and 4.43%, respectively, highlighting its potential for developing label-free electrochemical immunosensors for clinically relevant proliferation biomarkers. Full article

We report a highly sensitive label-free optical biosensor based on nematic liquid crystals, for the detection of proteins. The principles of biosensing are based on the change in the liquid crystal alignment induced by biomolecules adsorbed on the cell inner surface, which can be easily detected with a polarizing optical microscope. Although this approach is well-known, we propose here an experimental strategy that allows us to reach a detection limit of the order of 10⁻¹³ g/mL, orders of magnitude higher than the one reported in the literature for similar biosensors. Furthermore, our method leads to assessing a well-defined, specific dependence of protein concentration on cell birefringence, for rapid quantitative biosensing. The proposed biosensor can additionally be used for the detection of antibodies. Full article

Microbial imbalance and the spread of pathogenic microorganisms pose severe threats to human health and ecological security. Traditional microbial detection methods suffer from several drawbacks such as long detection time, low sensitivity, and insufficient specificity. As an emerging fluorescent probe, carbon dots (CDs) offer an innovative direction for microbial labeling and detection due to their ultra-small particle size, unique optical properties, excellent biocompatibility, and facile surface modifiability. Herein, this article reviews the research progress of CDs on microbial labeling and detection. The content covers a brief introduction of CDs and explores the main recognition strategies including non-covalent interactions and biomolecule-mediated targeted binding. It also elaborates on the application status of multi-modal sensing technologies for microbial detection, such as CDs-based fluorescent sensing, electrochemical sensing, and surface-enhanced Raman scattering (SERS) sensing. Additionally, the challenges faced in current research, such as achieving simultaneous detection of multiple pathogens and in vivo dynamic tracking, are analyzed, and the development prospects of CDs in fields like clinical diagnosis and public health monitoring are prospected. This review aims to provide comprehensive references for further research and application of CDs in the field of microbial detection. Full article

Label-free imaging is becoming ever more important, especially in modern molecular biophysics. This method allows observation of biological structures and dynamics without the alteration caused by dyes or genetic labels. Coherent Anti-Stokes Raman Scattering (CARS) microscopy represents a unique method that utilizes the intrinsic vibrational signatures of biomolecules, thereby transforming the field. Fluorescence-based methods show marked sensitivity, but may cause photobleaching, labeling artifacts, and inadequate biochemical detection. CARS enables chemically specific, real-time imaging of molecular structures, e.g., lipids, proteins and nucleic acids, within their natural environment. Over the past decade, advances in laser technology, detection methods, and computer analysis have turned CARS from a rare optical phenomenon into a useful tool applied in many fields, from basic research on molecular structure to practical biomedical imaging. This review presents the principles of CARS microscopy and the latest achievements in this field, highlighting its impact on molecular and cellular biophysics, as well as exploring the potential of artificial intelligence and multimodal approaches to increase its applications in precision medicine. In this context, CARS serves both a state-of-the-art imaging technique and a means of transforming internal molecular vibrations into information useful in biology and biophysics. In this way, it combines the physical sciences with molecular biology, enabling innovative biomedical research. Full article

Aptamers are powerful tools for detecting and analyzing biomolecules that consist of proteins or nucleic acids. However, their application to aptamers against viroids—highly structured self-replicating RNAs—has not yet been explored. In this study, a magnetic bead- and high-throughput sequencing-based SELEX (MB-HTS-SELEX) protocol for selecting potential aptamers against potato spindle tuber viroid (PSTVd) is presented. Full-length biotinylated-PSTVd RNA was transcribed in vitro, immobilized on streptavidin-coated magnetic beads, and incubated with a library of ~3.32 × 10¹⁴ molecules of random single-stranded oligo-DNAs (oligo-ssDNAs) of 20, 30, or 40 nucleotides (L20, L30, or L40, respectively) flanked by primer binding sites for downstream PCR amplification. Simultaneous biotin labeling of the anti-aptamer strand of the resulting double-stranded DNA (dsDNA) amplicons facilitated strand separation using streptavidin-coated magnetic beads. After 10 selection rounds, high-throughput sequencing, followed by bioinformatics analysis of the generated sequences, allowed for the detection of several enriched sequences, representing putative PSTVd-binding aptamers. Subsequent pull-down assays showed that the most abundant oligo-ssDNA in L30 was docked on PSTVd molecules. This combination method may ameliorate the selection of high-affinity aptamers against PSTVd, reduce the number of selection cycles, time, and other costs of aptamer production, thereby promoting future massive and cost-effective viroid detection and characterization. Full article

The cycloaddition reaction is one of the most common reactions in organic chemistry. It has been applied in various fields. Herein, we focus on the application of cycloaddition reactions in investigating biological molecules and materials using magnetic resonance techniques. To facilitate magnetic resonance studies such as electron paramagnetic resonance (EPR) spectroscopy and paramagnetic nuclear magnetic resonance (NMR) spectroscopy, there is often a requirement to attach spin labels and paramagnetic tags to the system of interest. The cycloaddition reaction is one of the ways to tether these spin labels and paramagnetic tags. In this review, we highlight the applications of various cycloaddition reactions such as the Cu(I)-catalyzed azide–alkyne cycloaddition (CuAAC) reaction, the strain-promoted azide–alkyne cycloaddition (SPAAC) reaction and the Diels–Alder reaction in the interdisciplinary field of magnetic resonance studies of biomolecules, including proteins, nucleic acids, carbohydrates, lipids and glycans, as well as materials. Full article

Mass spectrometry imaging (MSI) is an innovative analytical technique that integrates chemical analysis with spatial localization, enabling label-free, in situ detection and visualization of diverse biomolecules within tissue sections. This review summarizes the recent advances in the application of MSI to neurological disorders, with a focus on Parkinson’s disease, Alzheimer’s disease, schizophrenia, and traumatic brain injury. Studies have demonstrated that MSI can delineate the spatial heterogeneity of disease-related molecules—such as neurotransmitters, lipids, and metabolites—thereby providing new perspectives for understanding the pathological mechanisms of neurodegenerative and psychiatric diseases. Platforms including MALDI-MSI and DESI-MSI have been effectively employed for visualizing drug distribution, characterizing lipid metabolic pathways, and identifying spatial biomarkers. Although challenges remain in quantitative accuracy, spatial resolution, and the detection of low-abundance molecules, advances in high-resolution mass spectrometry, single-cell-level imaging, and multi-omics integration are expected to further enhance the utility of MSI in the investigation of brain diseases. Full article

The ring-assisted Mach–Zehnder interferometer (RA-MZI) has high sensitivity and fast optical response time, and it has been used as a label-free nanophotonic biosensor. Most RA-MZI-based biosensors, however, require chemical modification of the ring surface to immobilize biomolecules that can interact with target molecules for sensing. Here, we report a novel microfluidic-integrated RA-MZI (μFRA-MZI) where a microfluidic channel was fabricated right above the photonic ring resonator. μFRA-MZI allows for direct sample delivery to the RA-MZI without chemical modification of the ring surface and measures shifts in the resonance wavelength induced by the presence of target molecules, enabling label-free detection. In order to optimize the sensitivity of μFRA-MZI, seven devices were fabricated with varied design parameters, including the gap distance between the ring and the bus waveguide (G_ring), the length of the multi-mode interferometer (L_MMI), and the length of the directional coupler (L_DC). Photonic characterization showed that the device with G_ring = 1.2 μm, L_MMI = 15.5 μm, and L_DC = 13.5 μm exhibited the highest extinction ratio (ER) compared to the other six devices, consistent with the simulation-optimized design. Testing with NaCl solutions of varying concentrations yielded a bulk sensitivity of 11.48 nm/refractive index unit (RIU) and an ER of 0.41. With the potential to further improve the device’s sensitivity and the ability to detect samples directly in flow without chemical modifications of the ring resonator, μFRA-MZI will provide a robust and effective approach for label-free biosensing. Full article

ZnO nanostructures have attracted attention as transducer materials in optical biosensing platforms due to their wide bandgap, defect-mediated photoluminescence, high surface-to-volume ratio, and tunable morphology. This review examines how the dimensionality of ZnO nanostructures affects biosensor performance, particularly in terms of charge transport, signal transduction, and biomolecule immobilization. The synthesis approaches are discussed, highlighting how they influence crystallinity, defect density, and surface functionalization potential. The impact of immobilization strategies on sensor stability and sensitivity is also assessed. The role of ZnO in various optical detection schemes, including photoluminescence, surface plasmon resonance (SPR), localized (LSPR), fluorescence, and surface-enhanced Raman scattering (SERS), is reviewed, with emphasis on label-free and real-time detection. Representative case studies demonstrate the detection of clinically and environmentally relevant targets, such as glucose, dopamine, cancer biomarkers, and SARS-CoV-2 antigens, with limits of detection in the pico- to femtomolar range. Recent developments in ZnO-based hybrid systems and their integration into fiber-optic and microfluidic platforms are explored as scalable solutions for portable, multiplexed diagnostics. The review concludes by outlining current challenges related to reproducibility, long-term operational stability, and surface modification standardization. This work provides a framework for understanding structure–function relationships in ZnO-based biosensors and highlights future directions for their development in biomedical and environmental monitoring applications. Full article

In this work, we used a label-free biosensor that provides optical readouts to perform continuous detection of human interleukin 8 (IL-8), which is especially overexpressed in certain cancers and, thus, could be an effective biomarker for cancer prognosis estimation and therapy evaluation. For this purpose, we engineered a compact, portable, and easy-to-assemble biosensing module device. It combines a fluidic chip for reagent flow, a biosensing chip for signal transduction, and an optical readout head based on fiber optics in a single module. The biosensing chip is based on independent arrays of resonant nanopillar transducer (RNP) networks. We integrated the biosensing chip with the RNPs facing down in a simple and rapidly fabricated polydimethyl siloxane (PDMS) microfluidic chip, with inlet and outlet channels for the sample flowing through the RNPs. The RNPs were vertically oriented from the backside through an optical fiber mounted on a holder head fabricated ad hoc on polytetrafluoroethylene (PTFE). The optical fiber was connected to a visible spectrometer for optical response analysis and consecutive biomolecule detection. We obtained a sensogram showing anti-IL-8 immobilization and the specific recognition of IL-8. This unique portable and easy-to-handle module can be used for biomolecule detection within minutes and is particularly suitable for in-line sensing of physiological and biomimetic organ-on-a-chip systems. Cancer biomarkers’ continuous monitoring arises as an efficient and non-invasive alternative to classical tools (imaging, immunohistology) for determining clinical prognostic factors and therapeutic responses to anticancer drugs. In addition, the multiplexed layout of the optical transducers and the simplicity of the monolithic sensing module yield potential high-throughput screening of a combination of different biomarkers, which, together with other medical exams (such as imaging and/or patient history), could become a cutting-edge technology for further and more accurate diagnosis and prediction of cancer and similar diseases. Full article

Mid-infrared molecular sensing faces challenges in simultaneously achieving high-resolution qualitative identification and quantitative analysis of multiple biomolecules. To address this, we present a tunable mid-infrared sensing platform, integrating the simulation of a single-layer graphene square-aperture array sensor with a machine learning algorithm called principal component analysis for advanced spectral processing. The graphene square-aperture structure excites dynamically tunable localized surface plasmon resonances by modulating the graphene’s Fermi level, enabling precise alignment with the vibrational fingerprints of target molecules. This plasmon–molecule coupling amplifies absorption signals and serves as discernible “molecular barcodes” for precise identification without change in the structural parameters. We demonstrate the platform’s capability to detect and differentiate carbazole-based biphenyl molecules and protein molecules, even in complex mixtures, by systematically tuning the Fermi level to match their unique vibrational bands. More importantly, for mixtures with unknown total amounts and different concentration ratios, the principal component analysis algorithm effectively processes complex transmission spectra and presents the relevant information in a simpler form. This integration of tunable graphene plasmons with machine learning algorithms establishes a label-free, multiplexed mid-infrared sensing strategy with broad applicability in biomedical diagnostics, environmental monitoring, and chemical analysis. Full article