Detection Of Multiple Biomarkers Research Articles

Advances in Nanomaterial-Based Sensors for Early Disease Detection

Nanomaterial-based sensors are emerging as highly promising tools for the early detection of diseases, offering superior sensitivity, specificity, and miniaturization compared to traditional diagnostic methods. By leveraging the unique properties of nanomaterials such as gold nanoparticles, carbon nanotubes, graphene, and quantum dots, these sensors can detect disease biomarkers at much lower concentrations, enabling earlier diagnosis and intervention. This paper reviews the latest advancements in nanomaterial-based sensors, focusing on their applications in cancer, infectious diseases, and neurological disorders. Key developments include gold nanoparticle-based colorimetric sensors for cancer biomarker detection, graphene-based electrochemical sensors for prostate cancer and other malignancies, and quantum dot-based fluorescence sensors for both cancer and pathogen detection. Despite their promise, challenges remain, including issues with sensitivity, selectivity, reproducibility, biocompatibility, and scalability. Future directions for nanomaterial-based sensors involve improving their performance, developing multiplexed platforms for simultaneous detection of multiple biomarkers, and integrating these sensors into wearable or point-of-care devices. Overcoming these challenges and achieving regulatory approval will be crucial for the widespread clinical adoption of these advanced sensors, ultimately transforming healthcare by enabling more effective, rapid, and accessible early disease detection.

Highly sensitive multiplexed colorimetric lateral flow immunoassay by plasmon-controlled metal-silica isoform nanocomposites: PINs.

Lateral flow assay (LFA) systems use metal nanoparticles for rapid and convenient target detection and are extensively studied for the diagnostics of various diseases. Gold nanoparticles (AuNPs) are often used as probes in LFAs, displaying a single red color. However, there is a high demand for colorimetric LFAs to detect multiple biomarkers, requiring the use of multicolored NPs. Here, we present a highly sensitive multiplexed colorimetric lateral flow immunoassay by multicolored Plasmon-controlled metal-silica Isoform Nanocomposites (PINs). We utilized the localized surface plasmon resonance effect to create multi-colored PINs by precisely adjusting the distance between the NPs on the surface of PINs through the controlled addition of reduced gold and silver precursors. Through simulations, we also confirmed that the distance between nanoparticles on the surface of PINs significantly affects the color and colorimetric signal intensity of the PINs. We achieved multicolored PINs that exhibit stronger colorimetric signals, offering a new solution for LFA detection with high sensitivity and a 33 times reduced limit of detection (LOD) while maintaining consistent size deviations within 5%. We expect that our PINs-based colorimetric LFA will facilitate the sensitive and simultaneous detection of multiple biomarkers in point-of-care testing.

Activatable Multiplexed 19F NMR Probes for Dynamic Monitoring of Biomarkers Associated with Cellular Senescence.

Simultaneous detection of multiple biomarkers is crucial to achieve specific and dynamic analysis of cellular senescence, given its intrinsic high heterogeneity. Current approaches for senescence detection largely rely on fluorescence imaging, but fluorescent probes inevitably suffer from issues including autofluorescence and spectral overlap when being applied for the simultaneous detection of multiple biomarkers. Herein, we report an alternative strategy and design activatable multiplexed senoprobes based on 19F NMR for dynamic monitoring of cellular senescence. Differing from previous approaches, our strategy has two unique advantages. First, this strategy utilizes the changes in the 19F chemical shift as the signal output, which features by its fingerprint and quantifiable characters, thereby significantly enhancing the detection throughput toward biomarkers with minimized spectral overlapping. Second, the background signal is minimized, benefiting from the extremely low abundance of F in biological samples, and the detection accuracy can thus be improved. As a proof of concept, two activatable 19F NMR molecular probes are synthesized that specially respond to two key senescence-associated biomarkers (β-gal and ROS) and have been successfully demonstrated for dynamical and quantitative assessment of the changes of these biomarkers in different cellular models of senescence, without causing obvious cytotoxicity. Owing to the flexible molecular design, this work may offer a useful platform to create diversified 19F NMR senoprobes for deep understanding of cellular senescence across a wide range of aging-related diseases.

Acoustic Streaming Tunnel Enables Particle Velocity Stretching in Multiplex Flow Cytometry.

Multiplexed flow cytometry, known for its powerful high-throughput identification capability, is widely applied across various biomedical and clinical fields. However, classical flow cytometry relies on multichannel lasers and detectors, which are significant in cost and size, limiting their application in miniaturized assays. Herein, we developed an acoustic streaming-based flow cytometry technique that focuses on multisized microbeads flowing sheathlessly. This method enables the discrimination of particle types and the quantification of target protein concentrations using only a single detector. Microbeads of different sizes exhibit distinct behaviors in the continuous acoustic streaming tunnel, leading to an increased velocity difference during their transition under the laser spot. Consequently, a size detection method based on "velocity stretching" has been established. A multiplex assay of three proteins: cardiac troponin I, creatine kinase-MB and myoglobin, in acute myocardial infarction is performed to validate the feasibility and evaluate the performance of the system. This new multiplexed flow cytometry strategy is expected to enable low-cost and onsite detection of multiple biomarkers.

A fluorescent platform integrated with a “one-pot” nicking endonuclease signal amplification and magnetic separation for simultaneous detection of tumor markers

Although Enzyme-linked immunosorbent assay (ELISA) has been widely used for biomedical research, simultaneous sensitive and cost-effective detection of multiple biomarkers is challenging. Herein, we proposed a “one-pot” nicking endonuclease signal amplification (NESA)-based fluorescent aptasensor for simultaneous detection of carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP). Firstly, two aptamers were synchronously immobilized on the surface of magnetic nanoparticles (MNPs) by coupling with two complementary DNA (cDNA). CEA and AFP specifically recognized the aptamers and then the released cDNA (ssDNA) from the double-strands (dsDNA) triggers NESA, further breaking two detection probes which were labeled with the fluorescent dye (FAM and ROX) and its quencher (BHQ1 and BHQ2) at the same time. Then, the fluorescence signal of FAM and ROX were restored separately. The results indicated that the fluorescence intensity at the emission wavelength of 518 nm and 610 nm had a positive correlation with CEA and AFP concentrations, respectively. Under the optimum conditions, wider liner range of 1–500 ng mL−1 to CEA and 5–800 ng mL−1 to AFP of this fluorescent aptasensor were successfully obtained, achieving a detection limit of CEA and AFP were 0.7 ng mL−1 and 2 ng mL−1, respectively. Hence, it turned out that the aptasensor strategy can be a promising candidate for developing a newly fluorescence assay for the simultaneous quantitative detection of multiple tumor markers in matrix samples by changing the corresponding sequences of aptamer and fluorescent signal probe, which has great potential for the screening of early cancer.

Sensitive phenotyping of serum extracellular vesicles on a SERS-microfluidic platform for early-stage clinical diagnosis of ovarian carcinoma

Ovarian carcinoma (OvCa) poses a severe threat to women's health due to its high mortality rate and lack of efficient early diagnosis approach. There is evidence to suggest that nanosized small extracellular vesicles (sEVs) which carrying cell-specific components from OvCa can serve as potential diagnostic biomarkers. Herein, we reported a Surface-enhanced Raman Scattering (SERS)-multichannel microchip for sEVs (S-MMEV) assay to investigate the phenotype changes of sEVs. The microchip composed of seven microchannels, which enabled the parallel detection of multiple biomarkers to improve the detection accuracy. Using SERS probes conjugated with antibodies recognizing different biomarkers including ubiquitous EV biomarkers (i.e., tetraspanins; CD9, CD81) and putative OvCa tumor biomarkers (i.e. EpCAM, CD24, CA125, EGFR), we successfully analyzed the phenotypic changes of sEVs and accurately differentiated OvCa patients from healthy controls, even at early stage (I-II), with high sensitivity, high specificity and an area under the curve value of 0.9467. Additionally, the proposed approach exhibited higher sensitivity than conventional methods, demonstrating the efficiency of precise detection from cell culture and clinical samples. Collectively, the developed EV phenotyping approach S-MMEV could serve as a potential tool to achieve the early clinical diagnosis of OvCa for further precise diagnosis and personal treatment monitoring.

Digitalization of Enzyme-Linked Immunosorbent Assay with Graphene Field-Effect Transistors (G-ELISA) for Portable Ferritin Determination.

Herein, we present a novel approach to quantify ferritin based on the integration of an Enzyme-Linked Immunosorbent Assay (ELISA) protocol on a Graphene Field-Effect Transistor (gFET) for bioelectronic immunosensing. The G-ELISA strategy takes advantage of the gFET inherent capability of detecting pH changes for the amplification of ferritin detection using urease as a reporter enzyme, which catalyzes the hydrolysis of urea generating a local pH increment. A portable field-effect transistor reader and electrolyte-gated gFET arrangement are employed, enabling their operation in aqueous conditions at low potentials, which is crucial for effective biological sample detection. The graphene surface is functionalized with monoclonal anti-ferritin antibodies, along with an antifouling agent, to enhance the assay specificity and sensitivity. Markedly, G-ELISA exhibits outstanding sensing performance, reaching a lower limit of detection (LOD) and higher sensitivity in ferritin quantification than unamplified gFETs. Additionally, they offer rapid detection, capable of measuring ferritin concentrations in approximately 50 min. Because of the capacity of transistor miniaturization, our innovative G-ELISA approach holds promise for the portable bioelectronic detection of multiple biomarkers using a small amount of the sample, which would be a great advancement in point-of-care testing.

Grating Bio-Microelectromechanical Platform Architecture for Multiple Biomarker Detection.

A label-free biosensor based on a tunable MEMS metamaterial structure is proposed in this paper. The adopted structure is a one-dimensional array of metamaterial gratings with movable and fixed fingers. The moving unit of the optical detection system is a component of the MEMS structure, driven by the surface stress effect. Thus, these suspended optical nanoribbons can be moved and change the grating pattern by the biological bonds that happened on the modified cantilever surface. Such structural variations lead to significant changes in the optical response of the metamaterial system under illuminating angled light and subsequently shift its resonance wavelength spectrum. As a result, the proposed biosensor shows appropriate analytical characteristics, including the mechanical sensitivity of Sm = 11.55 μm/Nm-1, the optical sensitivity of So = Δλ/Δd = 0.7 translated to So = Δλ/Δσ = 8.08 μm/Nm-1, and the quality factor of Q = 102.7. Also, considering the importance of multi-biomarker detection, a specific design of the proposed topology has been introduced as an array for identifying different biomolecules. Based on the conducted modeling and analyses, the presented device poses the capability of detecting multiple biomarkers of disease at very low concentrations with proper precision in fluidic environments, offering a suitable bio-platform for lab-on-chip structures.

Analysis of cancer detection methods based on nanomaterials

This research discusses the application of nanomaterials in cancer detection and the comparison between various application methods, highlighting their high sensitivity and specificity in the detection of cancer biomarkers. Nanomaterials have the ability to selectively target and bind specific molecules, resulting in improved detection accuracy. They also facilitate the simultaneous detection of multiple biomarkers and enable non-invasive tumor visualization by various imaging methods. Despite the many advantages, there are still many challenges related to safety considerations and optimization of detection techniques that need to be addressed urgently. Nonetheless, the nanomaterials-based approach holds significant promise for improving cancer detection technology and improving early diagnosis. Further research is essential in order to fully exploit the power of nanomaterials in accurate and efficient cancer detection. By addressing these challenges, difficulties, and conducting additional research, nanomaterials have the potential to revolutionize cancer detection, resulting in improved results accuracy, earlier detection of disease, and better treatment for the benefit of humanity.

"One-Pot" Readout Cyano-Programmable SERS-Encoded Platform Enables Ultrasensitive and Interference-Free Detection of Multitarget Bioamines.

Surface-enhanced Raman spectroscopy (SERS) detection platforms with high signal-to-noise ratio in the "biological-silent" region (1800-2800 cm-1) are presently being developed for sensing and imaging applications, overcoming the limitations of traditional SERS studies in the "fingerprint" region. Herein, a series of cyano-programmable Raman reporters (RRs) operating in the "biological-silent" region were designed based on 4-mercaptobenzonitrile derivatives and then embedded in core-shell Au@Ag nanostars using a "bottom-up" strategy to provide SERS enhancement and encapsulation protection. The approach enabled the "one-pot" readout interference-free detection of multiple bioamines (histamine, tyramine, and β-phenethylamine) based on aptamer-driven magnetic-induced technology. Three cyano-encoded SERS tags resulted in separate SERS signals for histamine, tyramine, and β-phenethylamine at 2220, 2251, and 2150 cm-1, respectively. A target-specific aptamer-complementary DNA competitive binding strategy allowed the formation of microscale core-satellite assemblies between Fe3O4-based magnetic beads and the SERS tags, enabling multiple SERS signals to be observed simultaneously under a 785 nm laser excitation laser. The LODs for detection of the three bioamines were 0.61 × 10-5, 2.67 × 10-5, and 1.78 × 10-5 mg L-1, respectively. The SERS-encoded platform utilizing programmable reporters provides a fast and sensitive approach for the simultaneous detection of multiple biomarkers, paving the way for routine SERS analyses of multiple analytes in complex matrices.

Ultrasensitive Electrochemiluminescence Imaging Detection of Multiple miRNAs in Single Cells with a Closed Bipolar Electrode Array Chip.

In situ sensitive detection of multiple biomarkers in a single cell was highly necessary for understanding the pathogenesis mechanism and facilitating disease diagnosis. Herein, a bipolar electrode (BPE)-electrochemiluminescence (ECL) imaging chip was designed for ultrasensitive in situ detection of multiple miRNAs in single cells based on a dual-signal amplification strategy. A single cell was trapped and lysed within the microtrap of the cathode chamber and an HCR amplification process and nanoprobes (Fc/DNA/Fe3O4) were introduced, leading to a large number of electroactive molecules (Fc) being modified on the surface. Under a suitable potential, Fc+ in the cathodic chamber was reduced to Fc and L-012 was oxidized in the anodic chamber according to the electric neutrality principle of the bipolar electrode system, resulting in the ECL signal recorded by EMCCD. Ascribed to the dual-signal amplification, sensitive visual detection of miRNA-21 and miRNA-155 in single cells was achieved. For MCF-7 cells, miRNA-21 and miRNA-155 were calculated to be 4385 and 1932 copies/cell (median), respectively. For HeLa cells, miRNA-21 and miRNA-155 were calculated to be 1843 and 1012 copies/cell (median), respectively. The comprehensive evaluation of two kinds of miRNA could effectively eliminate error signals, and the detection precision was improved by 10%.

Parallel optofluidic detection of multiple cardiac biomarkers for point-of-care testing applications

Point-of-care testing (POCT), which enables personalized, accurate, affordable, and accessible medical services, is indispensable in the diagnostics, screening, and treatment of disease, especially for emergency and severe cases. Considering the complexity of disease, multiplexed point-of-care (MPOC) testing is very important for the simultaneous detection of several biomarkers to improve accuracy and increase analysis throughput. In this work, a microfiber interferometer based multiplexed optofluidic (MIMO) chip sensor is proposed for multiplexed, quantitative detection of cardiac biomarkers. Four microfiber interferometers are suspended and fixed in four separate microscale sample channels of an imprinted polydimethylsiloxane (PDMS) chip. A drop of the sample is distributed into the four parallel channels automatically and reacts with the probe on the microfiber sensor surface specifically, which leads to the wavelength shift of the spectrum resulting from evanescent wave interactions with biomolecules. There microfiber interferometers are functionalized with myoglobin (Myo) antibody, cardiac troponin I (cTnI) antibody, and creatine kinase–myocardial band isoenzymes (CK–MB) antibody respectively and the rest microfiber interferometer is with no functionalization, which acts as the control channel to reduce noise. By using compact dedicated commercial optical modules, miniaturized and low cost experimental setup is organized and achieve four channel simultaneous measurement through time division multiplexing. Three cardiac biomarkers, Myo, cTnI, and CK–MB, are synchronously detected on the proposed MIMO chip sensor. In a standard buffer sample, the log-linear sensing range of CK–MB is 0.2 ∼ 200 ng/mL with detection limit of 3.7 pg/mL, the log-linear sensing range of Myo is 2 ∼ 2000 ng/mL with detection limit of 0.5 ng/mL, and the log-linear sensing range of cTnI 0.02 ∼ 20 ng/mL with detection limit of 2.6 pg/mL, which all perfectly cover the actual diagnostic limit of myocardial infarction. Furthermore, the MIMO chip sensor maintains excellent performance of both specificity and detection capabilities in more complex serum samples. In double-blind trials of multiple biomarker detection, the MIMO chip sensor exhibits comparable detection ability with a commercial kit and good resistance to environmental disturbance. The proposed MIMO chip sensor sensing system offers a low-cost, portable, self-driven, and miniaturized scheme for a multiplexed point-of-care testing device, promoting the feasibility and accessibility of personalized medical services and paving the way for clinical application and industrialization.

Tau, Glial Fibrillary Acidic Protein, and Neurofilament Light Chain as Brain Protein Biomarkers in Cerebrospinal Fluid and Blood for Diagnosis of Neurobiological Diseases.

Neurological damage is the pathological substrate of permanent disability in various neurodegenerative disorders. Early detection of this damage, including its identification and quantification, is critical to preventing the disease's progression in the brain. Tau, glial fibrillary acidic protein (GFAP), and neurofilament light chain (NfL), as brain protein biomarkers, have the potential to improve diagnostic accuracy, disease monitoring, prognostic assessment, and treatment efficacy. These biomarkers are released into the cerebrospinal fluid (CSF) and blood proportionally to the degree of neuron and astrocyte damage in different neurological disorders, including stroke, traumatic brain injury, multiple sclerosis, neurodegenerative dementia, and Parkinson's disease. Here, we review how Tau, GFAP, and NfL biomarkers are detected in CSF and blood as crucial diagnostic tools, as well as the levels of these biomarkers used for differentiating a range of neurological diseases and monitoring disease progression. We also discuss a biosensor approach that allows for the real-time detection of multiple biomarkers in various neurodegenerative diseases. This combined detection system of brain protein biomarkers holds significant promise for developing more specific and accurate clinical tools that can identify the type and stage of human neurological diseases with greater precision.

Parallel detection of multiple biomarkers in a point-of-care-competent device for the prediction of exacerbations in chronic inflammatory lung disease

Sudden aggravations of chronic inflammatory airway diseases are difficult-to-foresee life-threatening episodes for which advanced prognosis-systems are highly desirable. Here we present an experimental chip-based fluidic system designed for the rapid and sensitive measurement of biomarkers prognostic for potentially imminent asthma or COPD exacerbations. As model biomarkers we chose three cytokines (interleukin-6, interleukin-8, tumor necrosis factor alpha), the bacterial infection marker C-reactive protein and the bacterial pathogen Streptococcus pneumoniae—all relevant factors in exacerbation episodes. Assay protocols established in laboratory environments were adapted to 3D-printed fluidic devices with emphasis on short processing times, low reagent consumption and a low limit of detection in order to enable the fluidic system to be used in point-of-care settings. The final device demonstrator was validated with patient sample material for its capability to detect endogenous as well as exogenous biomarkers in parallel.

Flexible Wearable Electrochemical Sensors Based on AuNR/PEDOT:PSS for Simultaneous Monitoring of Levodopa and Uric Acid in Sweat.

As a facile substitute for the invasive technique of blood testing, wearable electrochemical sensors exhibit high potential for the noninvasive and real-time monitoring of biomarkers in human sweat. However, owing to enzyme specificity, the simultaneous detection of multiple biomarkers by enzymatic analysis is challenging. Moreover, sweat accumulation under sensors causes sweat contamination, which hinders real-time biomarker detection from sweat. This study reports the design and fabrication of flexible wearable electrochemical sensors containing a composite comprising Au nanorods (AuNRs) and poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) for the nonenzymatic detection of levodopa (LD) and uric acid (UA) in sweat. Each sensor was integrated with a flexible three-electrode system and a microfluidic patch for sweat sampling. AuNRs immobilized by PEG-doped PEDOT:PSS showed excellent analytical performance for LD and UA at different potentials. Thus, the newly fabricated sensors could detect LD and UA over a broad detection range with high sensitivity and showed a low limit of detection for both species. On-body assessments confirmed the ability of these sensors to simultaneously detect LD and UA in real time. Therefore, this study could open new frontiers in the fabrication of wearable electrochemical sensors for the pharmacokinetic profile tracking of LD and gout management.

Simultaneous Detection of Multiple Biomarkers by Peptide Nucleic Acids‐Based Triplex Molecular Beacon in a Nanopore

AbstractIn this study, we propose an enhanced nanopore sensing strategy that utilizes a peptide nucleic acid (PNA)‐based triplex molecular beacon sensor to achieve the simultaneous detection of multiple biomarkers with a high degree of sensitivity. The sensor is a triplex switch composed of a triplex‐forming DNA strand and an oligo‐arginine‐tagged PNA strand, serving as the target recognition moiety and signal output element, respectively. Upon target binding to the recognition element of the sensor, the PNA signal output strand is released and a hybrid complex of the target‐DNA recognition strand is formed simultaneously. Due to the positive charges carried by the PNA‐Arg strands, they could be driven through the nanopore under positive electric field, effectively eliminating interferences from co‐existing target‐DNA complexes. This approach enables label‐free, one‐step detection of targets without requiring complex treatments and procedures. Leveraging the modular properties of DNA recognition strand, this method can be applied universally, and here, we successfully demonstrate its application using three SARS‐CoV‐2 related biomarkers.

The CRISPR-Cas13a Gemini System for noncontiguous target RNA activation

Simultaneous multi-target detection and multi-site gene editing are two key factors restricting the development of disease diagnostic and treatment technologies. Despite numerous explorations on the source, classification, functional features, crystal structure, applications and engineering of CRISPR-Cas13a, all reports use the contiguous target RNA activation paradigm that only enables single-target detection in vitro and one-site gene editing in vivo. Here we propose a noncontiguous target RNA activation paradigm of Cas13a and establish a CRISPR-Cas13a Gemini System composed of two Cas13a:crRNA binary complexes, which can provide rapid, simultaneous, highly specific and sensitive detection of two RNAs in a single readout, as well as parallel dual transgene knockdown. CRISPR-Cas13a Gemini System are demonstrated in the detection of two miRNAs (miR-155 and miR-375) for breast cancer diagnosis and two small RNAs (EBER-1 and EBER-2) for Epstein-Barr virus diagnosis using multiple diagnostic platforms, including fluorescence and colorimetric-based lateral flow systems. We also show that CRISPR-Cas13a Gemini System can knockdown two foreign genes (EGFP and mCherry transcripts) in mammalian cells simultaneously. These findings suggest the potential of highly effective and simultaneous detection of multiple biomarkers and gene editing of multiple sites.

Visual and quantitative lateral flow immunoassay based on Au@PS SERS tags for multiplex cardiac biomarkers

Rapid and sensitive detection of multiple biomarkers by lateral flow immunoassay (LFIA) remains challenging for signal amplification for commonly used nanotags. Herein, we report a novel LFIA strip for visual and highly sensitive analysis of two cardiac biomarkers based on functionalized gold nanoparticles @ polystyrene microsphere (Au@PS）microcavity as surface-enhanced Raman scattering (SERS) tags. Antibody-modified Au@PS was designed as a SERS label. The evanescent waves propagating along the surface of the PS microcavity and the localized surface plasmons of the gold nanoparticles were coupled to enhance the light-matter interaction synergistically for Raman signal enhancement. In this strategy, the proposed Au@PS SERS tags-based LFIA was carried out to quantify the content of the heart failure and infarct biomarkers synchronously within 15 min and get the limits of detection of 1 pg/mL and 10 pg/mL for cardiac troponin I (cTnI) and N-terminal natriuretic peptide precursor (NT-proBNP), respectively. The results demonstrated 10–20 folds more sensitivity than that of the standard colloidal gold strip and fluorescent strip for the same biomarkers. This novel quantitative LFIA shows promise as a high-sensitive and visual sensing method for relevant clinical and forensic analysis.

Ultrasensitive and Multiple Biomarker Discrimination for Alzheimer's Disease via Plasmonic & Microfluidic Sensing Technologies.

As the population ages, the worldwide prevalence of Alzheimer's disease (AD) as the most common dementia in the elderly is increasing dramatically. However, a long-term challenge is to achieve rapid and accurate early diagnosis of AD by detecting hallmarks such as amyloid beta (Aβ42). Here, a multi-channel microfluidic-based plasmonic fiber-optic biosensing platform is established for simultaneous detection and differentiation of multiple AD biomarkers. The platform is based on a gold-coated, highly-tilted fiber Bragg grating (TFBG) and a custom-developed microfluidics. TFBG excites a high-density, narrow-cladding-mode spectral comb that overlaps with the broad absorption of surface plasmons for high-precision interrogation, enabling ultrasensitive monitoring of analytes. In situ detection and in-parallel discrimination of different forms of Aβ42 in cerebrospinal fluid (CSF) are successfully demonstrated with a detection of limit in the range of ≈30-170 pg mL-1, which is one order of magnitude below the clinical cut-off level in AD onset, providing high detection sensitivity for early diagnosis of AD. The integration of the TFBG sensor with multi-channel microfluidics enables simultaneous detection of multiple biomarkers using sub-µL sample volumes, as well as combining initial binding rate and real-time response time to differentiate between multiple biomarkers in terms of binding kinetics. With the advantages of multi-parameter, low consumption, and highly sensitive detection, the sensor represents an urgently needed potentials for large-scale diagnosis of diseases at earlystage.

Recent Advances in Fluorescent Probes for Cancer Biomarker Detection.

Many important biological species have been identified as cancer biomarkers and are gradually becoming reliable targets for early diagnosis and late therapeutic evaluation of cancer. However, accurate quantitative detection of cancer biomarkers remains challenging due to the complexity of biological systems and the diversity of cancer development. Fluorescent probes have been extensively utilized for identifying biological substances due to their notable benefits of being non-invasive, quickly responsive, highly sensitive and selective, allowing real-time visualization, and easily modifiable. This review critiques fluorescent probes used for detecting and imaging cancer biomarkers over the last five years. Focuses are made on the design strategies of small-molecule and nano-sized fluorescent probes, the construction methods of fluorescence sensing and imaging platforms, and their further applications in detection of multiple biomarkers, including enzymes, reactive oxygen species, reactive sulfur species, and microenvironments. This review aims to guide the design and development of excellent cancer diagnostic fluorescent probes, and promote the broad application of fluorescence analysis in early cancer diagnosis.