Label-free Detection System Research Articles

Dealing with plasmonic crystal biosensors: Sensitivity assessment of nanodisks/nanoholes arrayed plasmonic system for label-free DNA detection

Label-free optical deoxyribonucleic acid (DNA) sensing with arrayed plasmonic nanostructures (plasmonic crystals) is a promising technology for biomedical diagnosis and bioanalytical science. Plasmonic biosensors can detect target biomolecules by utilizing the shift in plasmonic resonance caused by changes in the surrounding refractive index (RI) attributed to the capture of target biomolecules using a recognizer. Conventional explanations for the sensitivity of plasmonic crystals are based on bulk (BRIS) and surface RI sensitivities (SRIS) for basic plasmonic nanoparticles despite their unique properties such as surface lattice resonances (SLRs), wherein localized surface plasmons (LSPs) cooperatively oscillate with their pitch. Therefore, investigating the sensitivity of SLRs is imperative for improving sensing performance. In this study, the sensitivity of adenomatous polyposis coli (APC) gene-related DNA hybridization detection of complementary plasmonic crystals composed of nanodisks (PNDs) on or under plasmonic nanoholes (PNHs) was investigated considering the SLR properties. The BRIS was measured using the conventional definition of the peak wavelength shift per unit RI increment (nm/RIU) followed by the SRIS measurement using the layer-by-layer method. The BRIS and SRIS measurements reflect the practical sensitivity for DNA detection. PNHs had higher sensitivity than PNDs, with a limit of detection of 0.30 nM. Further, only the SLR-based mode responded to localized RI changes because of DNA hybridization, whereas both the LSPs- and SLR-based modes responded to uniform RI changes caused by layer-by-layer coating. Our investigation will open up possibilities and opportunities for plasmonic crystal biosensors.

Label-Free and Ultra-Sensitive Detection of Dexamethasone Using a FRET Aptasensor Utilizing Cationic Conjugated Polymers.

Dexamethasone (Dex) is a widely used glucocorticoid in medical practice, with applications ranging from allergies and inflammation to cerebral edema and shock. Despite its therapeutic benefits, Dex is classified as a prohibited substance for athletes due to its potential performance-enhancing effects. Consequently, there is a critical need for a convenient and rapid detection platform to enable prompt and accurate testing of this drug. In this study, we propose a label-free Förster Resonance Energy Transfer (FRET) aptasensor platform for Dex detection utilizing conjugated polymers (CPs), cationic conjugated polymers (CCPs), and gene finder probes (GFs). The system operates by exploiting the electrostatic interactions between positively charged CCPs and negatively charged DNA, facilitating sensitive and specific Dex detection. The label-free FRET aptasensor platform demonstrated robust performance in detecting Dex, exhibiting high selectivity and sensitivity. The system effectively distinguished Dex from interfering molecules and achieved stable detection across a range of concentrations in a commonly used sports drink matrix. Overall, the label-free FRET Dex detection system offers a simple, cost-effective, and highly sensitive approach for detecting Dex in diverse sample matrices. Its simplicity and effectiveness make it a promising tool for anti-doping efforts and other applications requiring rapid and accurate Dex detection.

Hyperspectral enhanced imaging analysis of nanoparticles using machine learning methods.

Nanoparticle (NP)-based technologies have gained significant attention in targeted drug delivery, encompassing chemotherapies, photodynamic therapy, and immunotherapy. Hyperspectral imaging (HSI) emerges as a label-free, minimally invasive, and high-throughput technique for quantitative NP analysis. Despite its growing importance, the application of HSI to nanoparticle analysis, especially for label-free characterization and classification, remains limited. Here, we propose a novel method integrating hyperspectral imaging with a spectral noise reduction method and machine learning (ML) for robust nanoparticle classification. There are many challenges to extracting information from noisy and overlapping particles in HSI data. To surmount these challenges, we propose a spectral angle matching (SAM) algorithm to effectively denoise hyperspectral datasets. Complementing this, we employ a support vector machine (SVM) algorithm for classification, leveraging preprocessed HSI data to extract unique spectral signatures. Our hyperspectral imaging classification of multiple nanoparticle types reveals distinct spectral characteristics inherent to each class. The classification accuracy reaches 99.9% for single nanoparticle types, highlighting the efficiency of our method. In the case of classifying multiple particle types, the overall accuracy also reaches 99.9%. Visualization of the NP classification map further demonstrates the efficacy of our model. The application of the SAM-SVM algorithm in hyperspectral analysis outperforms traditional SVM methods in classifying multiple samples, highlighting the potential of our nanoparticle analysis. Our findings not only address the challenges posed by noisy and overlapping particles but also demonstrate the potential of hyperspectral imaging in advancing real-time and label-free detection systems for diverse biomedical applications.

“One-to-many” signal-output strategy-based CRISPR/Cas12a system for sensitive label-free fluorescence detection of HBV-DNA

Although CRISPR/Cas12a systems significantly enhance the analytical accuracy and flexibility of fluorescent biosensors, their sensitivity is limited by traditional “one-to-one” mediation types and ineffective signal-output turnover routes. Herein, we demonstrate a “one-to-many” signal-output strategy-based CRISPR/Cas12a systems resembling a “seaweed” to enhance the sensitivity. Based on dendrimer DNA from high-dimensional hybridization chain (HCR) of three hairpin-free DNA building blocks, the 3D magnetic DNA machine was created. The HBV-DNA initiates the rolling circle amplification (RCA) reaction and produces DNA nanowires to activate the CRISPR/Cas12a system. The trans-cleavage of the “seaweed root” by CRISPR/Cas12a system left dendrimer DNA in solution, thus, adding SYBR Green I (SG I) to the high-density DNA duplexes, achieving multiple-turnover label-free fluorescence signal output demonstrated and a low LOD (1.502 pM). However, in the absence of target, the blocked RCA failed to activate the CRISPR/Cas12a system, resulting in complete separation from substrate and negligible fluorescence signals. Moreover, the mandatory RCA-based pre-amplification of the DNA activator could efficiently trigger the multiple-turnover trans-cleavage activity of Cas12a. it can cleave one single-stranded linker of “seaweed-like” DNA machine, thereby releasing massive DNA duplex-enriched dendrimer DNA with a “one-to-many” signal-output turnover. By coupling the periodically extended Cas12a activator generated by RCA with hyperbranched DNA duplex by high-dimensional HCR, compact 3D extension structures were formed, achieving high-density fluorescence distribution in focal volume, avoiding signal dilution and ensuring high enhancement. Additionally, spiked recoveries in physiological media exceeded 95%, demonstrating the potential application of such platforms in clinical diagnosis.

Smartphone-powered portable chemiresistive sensing system for label free detection of lead ions in water

Lead (Pb) is a toxic heavy metal that can contaminate water sources and cause serious health problems. Detecting Pb in water is essential for protecting public health and the environment. However, the current state of technology heavily depends upon traditional bench-top equipment for detecting toxic heavy metal ions dissolved in water. In this paper we report a portable, label-free chemiresistive sensor for the selective detection of Pb ions dissolved in water. Lithography patterned micro-ineterdigitated electrodes (µIDEs) modified with Graphene oxide (GO) is used as the transducing agent to capture the sensor’s electrical response on a portable readout platform with a smartphone interface. Graphene oxide functionalized with environment friendly oligosaccharide β- Cyclodextrin acts as the sensing material. The sensor’s electrical properties undergo variation in the presence of Pb ions, hence serving as a parameter for successfully detecting the targeted heavy metal ion. The designed sensor is highly selective towards Pb ions, even in the presence of other heavy metal ions, with the lowest detection concentration being 100 pico Molar (pM). The smartphone-powered portable readout circuit, along with the sensor, makes the entire system energy-efficient and cost-effective, providing a quick analysis of the sensor output. Further, an ML algorithm is used to enhance the data analysis and assist the classification based on sensor response.

Determination of uric acid in serum by SERS system based on VO-MnCo2O4/Ag nanozyme

The level of uric acid is crucial to human health. Octahedral oxygen vacancy MnCo2O4/Ag (VO-MnCo2O4/Ag) nanozyme was successfully prepared by simple hydrothermal, calcination and self-reduction methods. VO-MnCo2O4/Ag nanozyme is rich in Mn2+/Mn3+ and CO2+/CO3+ redox electron pairs, large specific surface area and oxygen vacancies. VO-MnCo2O4/Ag nanozyme showed high uricase-like activity and peroxidase-like activity. At the same time, the SERS signal of the detected molecule could be significantly enhanced after the catalytic reaction of the VO-MnCo2O4/Ag nanozyme. The Km values of VO-MnCo2O4/Ag nanozyme for H2O2 and TMB were 0.04 mM and 0.027 mM respectively. Based on the uric acid oxidase-like and peroxidase-like activities of VO-MnCo2O4/Ag, we developed a label-free, sensitive, and reliable SERS uric acid detection system. The detection linear range of uric acid is 0.01 μM–1000 μM and the detection of limit is 7.8 × 10−9 M. The results show that the sensing system has good accuracy, sensitivity, selectivity, and stability. It can be applied to the determination of samples under different conditions. This study provides profound insights into the design of enzyme-like activity regulation and SERS properties regulation of nanozymes, provides guidance for the study of reaction kinetics and catalytic mechanism of nanozymes, and has broad application prospects in the field of nanozymes and SERS sensing analysis.

Multi-dimensional plasmonic coupling system for efficient enrichment and ultrasensitive label-free SERS detection of bilirubin based on graphene oxide-Au nanostars and Au@Ag nanoparticles

Rapid and sensitive detection of free bilirubin (BR) is essential for early diagnosis of jaundice and other hepatobiliary diseases. Inspired by sandwich immunoassay strategy, a multi-dimensional plasmonic coupling SERS platform composed of graphene oxide-Au nanostars nanocomposites (GANS NCs) and Au@Ag nanoparticles (NPs) was designed for label-free detection of BR. Specifically, GANS NCs were first prepared, and their excellent SERS activity was ascribed to synergistic enhancement effect of electromagnetic enhancement and chemical enhancement. Furthermore, SERS spectroscopy was used to monitor the adsorption process of BR. Subsequently, secondary reinforcing Au@Ag NPs were directly added, ultimately resulting in a multi-dimensional plasmonic coupling effect. The SERS enhancing mechanism of coupled system was discussed through electromagnetic field simulations. Interestingly, the high-density hotspots generated by strong plasmonic coupling in GANS-Au@Ag substrate could lead to more extraordinary SERS enhancing behavior compared to GANS NCs. Sensing efficiency of the SERS platform was examined by BR with a detection limit down to 10−11 M. Besides, GANS-Au@Ag NCs performed high uniformity and reproducibility. This work not only opens up a new avenue for construction of multi-dimensional plasmonic coupling system, but also offers a new biosensing technology for label-free diagnosis of BR-related diseases, thereby expecting to be applied in clinical diagnosis.

A plug-and-play aptamer diagnostic platform based on linear dichroism spectroscopy.

A plug-and-play sandwich assay platform for the aptamer-based detection of molecular targets using linear dichroism (LD) spectroscopy as a read-out method has been demonstrated. A 21-mer DNA strand comprising the plug-and-play linker was bioconjugated onto the backbone of the filamentous bacteriophage M13, which gives a strong LD signal due to its ready alignment in linear flow. Extended DNA strands containing aptamer sequences that bind the protein thrombin, TBA and HD22, were then bound to the plug-and-play linker strand via complementary base pairing to generate aptamer-functionalised M13 bacteriophages. The secondary structure of the extended aptameric sequences required to bind to thrombin was checked using circular dichroism spectroscopy, with the binding confirmed using fluorescence anisotropy measurements. LD studies revealed that this sandwich sensor design is very effective at detecting thrombin down to pM levels, indicating the potential of this plug-and-play assay system as a new label-free homogenous detection system based on aptamer recognition.

Detection speed optimization of the OI-RD microscope for ultra-high throughput screening.

The oblique-incidence reflectivity difference (OI-RD) microscope is a label-free detection system for microarrays that has many successful applications in high throughput drug screening. The increase and optimization of the detection speed of the OI-RD microscope will enable it to be a potential ultra-high throughput screening tool. This work presents a series of optimization methods that can significantly reduce the time to scan an OI-RD image. The wait time for the lock-in amplifier was decreased by the proper selection of the time constant and development of a new electronic amplifier. In addition, the time for the software to acquire data and for translation stage movement was also minimized. As a result, the detection speed of the OI-RD microscope is 10 times faster than before, making the OI-RD microscope suitable for ultra-high throughput screening applications.

Validation of Selective Capture of Fimbriated Uropathogenic Escherichia coli by a Label-free Engineering Detection System Using Mannosylated Surfaces.

Label-free detection of pathogens is of major concern to the microbiologist community. Most procedures require several steps and amplification techniques. Carbohydrates are well-established receptors for host-pathogen interactions, which can be amplified using glycodendritic architectures on the basis of multivalent binding interactions. Given that uropathogenic Escherichia coli bacterial FimH is based on such mannopyranoside-binding interactions, we demonstrate herein that synthetic monomeric and trimeric thiolated α-d-mannosides can be effectively bound to gold substrate-functionalized self-assembled monolayers (SAMs) preactivated with maleimide functionalities. Mannosides grafted onto SAMs were followed using Quartz Crystal Microbalance with Dissipation (QCM-D). Binding recognition efficiency was first evaluated using the plant lectin from Canavalia ensiformis (ConA) also using QCM-D. We showed a direct correlation between the amount of mannoside bound and the lectin attachment. Even though there was less trimer bound (nM/cm2) to the surface, we observed a 7-fold higher amount of lectin anchoring, thus further demonstrating the value of the multivalent interactions. We next examined the relative fimbriated E. coli selective adhesion/capture to either the monomeric or the trimeric mannoside bound to the surface. Our results established the successful engineering of the surfaces to show E. coli adhesion via specific mannopyranoside binding but unexpectedly, the monomeric derivative was more efficient than the trimeric analog, which could be explained by steric hindrance. This approach strongly suggests that it could be broadly applicable to other Gram-negative bacteria sharing analogous carbohydrate-dependent binding interactions.

High-throughput "read-on-ski" automated imaging and label-free detection system for toxicity screening of compounds using personalised human kidney organoids.

Organoid models are used to study kidney physiology, such as the assessment of nephrotoxicity and underlying disease processes. Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies, but there is a need to accelerate basic and translational research in the field. Here, we developed an automated continuous imaging setup with the "read-on-ski" law of control to maximize temporal resolution with minimum culture plate vibration. High-accuracy performance was achieved: organoid screening and imaging were performed at a spatial resolution of 1.1 μm for the entire multi-well plate under 3 min. We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system. The acquired images were processed via machine learning-based classification and segmentation algorithms, and the toxicity in kidney organoids was determined with 95% accuracy. The results obtained by the automated "read-on-ski" imaging device, combined with label-free and non-invasive algorithms for detection, were verified using conventional biological procedures. Taking advantage of the close-to-in vivo-kidney organoid model, this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.

Label-free biological sample detection and non-contact separation system based on microfluidic chip.

The detection and separation of biological samples are of great significance for achieving accurate diagnoses and state assessments. Currently, the detection and separation of cells mostly adopt labeling methods, which will undoubtedly affect the original physiological state and functions of cells. Therefore, in this study, a label-free cell detection method based on microfluidic chips is proposed. By measuring the scattering of cells to identify cells and then using optical tweezers to separate the target cells, the whole process without any labeling and physical contact could realize automatic cell identification and separation. Different concentrations of 15 µm polystyrene microspheres and yeast mixed solution are used as samples for detection and separation. The detection accuracy is over 90%, and the separation accuracy is over 73%.

Rapid and facile electrospray preparation of CsPbBr3@PMMA fluorescent microspheres for fluorescent detection of ALP in biological samples

All-inorganic cesium lead halide perovskite quantum dots (QDs) have attracted more attention due to their excellent optoelectronic and photovoltaic properties. However, there are few applications in biochemical detection based on their unique photoluminescence property mainly because of the instability to external stimulus. In this work, we focus on a rapid preparation method of CsPbBr3@poly (methyl methacrylate) (PMMA) fluorescent composite microspheres based on electrospray to protect CsPbBr3 QDs from the external stimuli and stabilize them in aqueous environment. The CsPbBr3@PMMA composite microspheres effectively retained over 70% fluorescence intensity after being immersed in deionized water for 20 days. Based on the effective quenching of CsPbBr3@PMMA composite microspheres by MnO2 nanosheets, we developed a simple, label-free, low-cost, and facile fluorescent detection system for assay of alkaline phosphatase (ALP) in biological samples. The method shows a good linear relationship from 10.0 mU/mL to 100 mU/mL (R2 = 0.998) with the detection limit of 4.85 mU/mL and the recovery range of 95.1–110%. In summary, the PMMA coating can simply and effectively stabilize the CsPbBr3 QDs and preserve the luminescence of the CsPbBr3 QDs. The excellent applicability of the composite microspheres-based fluorescent detection system in human serum samples further demonstrates the potential application in the aqueous phase-based biochemical detection for clinical diagnosis and biomedical research.

Metal-Modified Montmorillonite as Plasmonic Microstructure for Direct Protein Detection.

Thanks to its negative surface charge and high swelling behavior, montmorillonite (MMT) has been widely used to design hybrid materials for applications in metal ion adsorption, drug delivery, or antibacterial substrates. The changes in photophysical and photochemical properties observed when fluorophores interact with MMT make these hybrid materials attractive for designing novel optical sensors. Sensor technology is making huge strides forward, achieving high sensitivity and selectivity, but the fabrication of the sensing platform is often time-consuming and requires expensive chemicals and facilities. Here, we synthesized metal-modified MMT particles suitable for the bio-sensing of self-fluorescent biomolecules. The fluorescent enhancement achieved by combining clay minerals and plasmonic effect was exploited to improve the sensitivity of the fluorescence-based detection mechanism. As proof of concept, we showed that the signal of fluorescein isothiocyanate can be harvested by a factor of 60 using silver-modified MMT, while bovine serum albumin was successfully detected at 1.9 µg/mL. Furthermore, we demonstrated the versatility of the proposed hybrid materials by exploiting their plasmonic properties to develop liquid label-free detection systems. Our results on the signal enhancement achieved using metal-modified MMT will allow the development of highly sensitive, easily fabricated, and cost-efficient fluorescent- and plasmonic-based detection methods for biomolecules.

Ultrasensitive label-free detection for lung cancer CYFRA 21-1 DNA based on ring-opening polymerization

Cytokeratin fragment antigen 21-1 (CYFRA 21-1) DNA is perceived as sensitive tumor marker for the diagnosis of non-small cell lung cancer and other tumor. Herein, linear chain poly(ε-caprolactone) (PCL) synthesized by ring-opening polymerization is applied to ultrasensitive label-free electrochemical impedance detection system for CYFRA 21-1 DNA. First, thiolated peptide nucleic acid (PNA) is self-assembled into the Au electrode surface through the formation of Au–S bonds, allowing the PNA to act as biomolecular probe and form PNA/DNA heteroduplex with the target DNA via specific hybridization. Then, PCL is conjugated to the immobilized DNA on the electrode via “carboxylate–Zr4+–phosphate” bridges. Finally, the electrochemical response of modified PNA/DNA/Zr4+/PCL electrode is determined by electrochemical impedance method to quantify of CYFRA 21-1 DNA. Under optimal conditions, this method exhibits highly sensitivity with a broad linear range (0.1 fM – 1 nM) (R2 = 0.995) and the limit of detection (LOD) is as low as 10.73 aM, which is equivalent to just 64 molecules in a 10 μL sample. What's more, the high selectivity, good anti-interference, label-free operation, and real-time monitoring in complex samples of the proposed strategy demonstrate its broad application for the early diagnosis and clinical monitoring.

High sensitivity label-free detection of Fe3+ ion in aqueous solution using fluorescent MoS2 quantum dots

MoS2 quantum dots were successfully prepared by a one-step hydrothermal exfoliation procedure. The prepared MoS2 quantum dots are monodispersed and have a circular shape, with an average diameter of 4.67 nm. Excited by UV-light, MoS2 quantum dots emit blue-purple fluorescence, and the fluorescence intensity is suppressed with the addition of Fe3+ ions. On this basis a novel fluorescent MoS2 quantum dots–Fe3+ label-free detection system is proposed. The decay curves of fluorescence show that the fluorescence quenching follows a static process. The binding stoichiometry ratio of Fe3+ ions toward MoS2 quantum dots is recognized as 1:1, based on the Benesi-Hildebrand and Job’s plot methods. For the concentration range of 0–50 μmol/L, the Fe3+ and fluorescence intensity are linearly related to 99.39%. The detection limit reaches 1 μmol/L. Several common ions were also examined that have little cross-influence on the fluorescence intensity of the MoS2 quantum dots. The fluorescent MoS2 quantum dots–Fe3+ probe has good performances and provides a novel, convenient and effective approach for the detection of Fe3+ of extremely low concentrations.

Immobilized Polydiacetylene Lipid Vesicles on Polydimethylsiloxane Micropillars as a Surfactin-Based Label-Free Bacterial Sensor Platform

Accurate detection and sensing of bacteria are becoming increasingly important not only in microbiology but in a variety of fields including medicine, food, public health, and environmental science. However, even new rapid methods are not convenient enough. Here, we describe a simple and efficient label-free bacterial detection system using the polydiacetylene (PDA) liposomes immobilized on the 3D polydimethylsiloxane (PDMS) micropillars. Our system utilizes the colorimetric response of amine functionalized PDA vesicles to surfactin, a bacterial cyclic lipopeptide commonly released by Gram-positive Bacillus species as an antibiotic. To improve the sensitivity of PDA vesicles to surfactin by increasing the number and surface area of immobilized vesicles, the PDA vesicles were immobilized on the micropillar structure to give a hierarchical 3D PDA vesicle structure. For the fabrication of the 3D micropillar structure, polydimethylsiloxane (PDMS) was used to overcome the limitations imposed by silicon-based fabrication. In contrast to the 2D-PDA-PDMS system, which could only hardly detect the presence of 500 µM surfactin, the 3D-PDA-PDMS system could efficiently detect the presence of 5 µM surfactin and the initial presence of 4 x 101 cells/ml of Bacillus subtilis NCIB3610, which actively produces surfactin. Furthermore, bacterial strains that are known to produce no surfactin, such as Staphylococcus aureus Newman, Escherichia coli DH5, and Pseudomonas aeruginosa PA14 were not detected by our system suggesting that the 3D-PDA-PDMS system is highly specific to surfactin but not to other chemicals produced by bacteria. Taken together, our results suggest that the 3D-PDA-PDMS system can sensitively and selectively be used for the high throughput detection and screening of biotechnologically important surfactin-producing bacterial strains.

Molecularly imprinted polymer based quartz crystal microbalance sensor system for sensitive and label-free detection of synthetic cannabinoids in urine

Herein, we prepared a novel quartz crystal microbalance (QCM) sensor for synthetic cannabinoids (JWH-073, JWH-073 butanoic acid, JWH-018 and JWH-018 pentanoic acid,) detection. Firstly, the synthetic cannabinoid (SCs) imprinted (MIP) and non-imprinted (NIP) nanoparticles were synthesized by mini-emulsion polymerization system. The SCs-imprinted nanoparticles were first characterized by SEM, TEM, zeta-size and FTIR-ATR analysis and then were dropped onto the gold QCM surface. The SCs-imprinted QCM sensor was characterized by an ellipsometer, contact angle, and AFM. The limit of detection was found as 0.3, 0.45, 0.4, 0.2 pg/mL JWH-018, JWH-073, JWH-018 pentanoic acid and JWH-073 butanoic acid, respectively. The selectivity of the SCs-imprinted QCM sensor was shown by using JWH-018, JWH-018 pentanoic acid, JWH-073 and JWH-073 butanoic acid. According to the results, the SCs-imprinted QCM sensors show highly selective and sensitive in a broad range of synthetic cannabinoid concentrations (0.0005–1.0 ng/mL) in both aqueous and synthetic urine solutions.

Detection of heart-type fatty acid-binding protein (h-FABP) using piezoresistive polymer microcantilevers functionalized by a dry method

Piezoresistive microcantilever-based sensor platform is being used for the last two decades due to their low cost, rapid response and label-free detection system. In this work, we are reporting a microfabricated piezoresistive SU-8/carbon black (polymer cantilever)-based sensor platform for the detection of a clinically important early-stage cardiac marker, i.e., fatty acid-binding protein. It is a most preferred cardiac marker for the diagnosis of acute myocardial infarction. The embodiment of the sensor is a SU-8 microcantilever chip with an integrated nanoparticle composite (carbon black) as a piezoresistor for on-chip electrical transduction. Prior to improving the sensing and susceptibility towards the specific target biomolecule (i.e., h-FABP), the fabricated SU-8 polymer cantilevers were subjected to tailored functionalization. This includes the use of an in-house dry method of hot wire chemical vapour deposition technique to graft amine groups onto the SU-8 surface. The surface-modified microcantilevers were further integrated with a polydimethylsiloxane liquid flow cell and connected externally with an electrical read-out system. Immobilization of the antibody corresponding to the marker protein on the microcantilever surface and subsequent recording of the signal generated upon the antibody–antigen interaction were carried out inside the liquid flow cell. Using our optimized immobilization protocol with this experimental set-up, we were successfully able to detect h-FABP concentration as low as 100 ng/ml.

Microfluidic multi-angle laser scattering system for rapid and label-free detection of waterborne parasites.

A microfluidic laser scattering system for rapid and label-free detection of single waterborne parasites in microfluidic flows was designed, fabricated and demonstrated. The key novelty of the system lies in the integration of functional modules involving pre-concentration, on-chip laser scattering detection, and pattern recognition. The silicon-based pre-concentration chip can concentrate 10 ml reagent water sample spiked with protozoa (oo)cysts into a volume of 200 μl in ~30 minutes. The concentrated sample is further channeled into the on-chip laser scattering detection module at a flow rate of 10 μl/min, which can collect the multi-angle scattering pattern of single flowing microparticles. The Zernike moment features of scattering patterns are extracted using principal component analysis, and classification of scattering patterns are performed using the linear discriminator analysis algorithm. As a result, Cryptosporidium parvum oocysts and Giardia lamblia cysts spiked in ~10 ml reagent water can be enumerated and identified within an hour without labeling, with a mean recovery efficiency of ~73% and average accuracies of 96%, 97%, 97% and 98% at concentrations of 10, 50, 100, 300 (oo)cysts per 10 ml water, respectively. We believe that this compact microfluidic laser scattering system has potential for rapid and label-free water quality monitoring in field and resource-limited environments.