Dynamic Light Scattering Signal Research Articles

Ultrasensitive dynamic light scattering immunodetection of alpha-fetoprotein using heptamer-amplified nanoparticle crosslinking aggregation.

Anovel construction strategy is introducedfor an ultrasensitive dynamic light scattering (DLS) immunosensor targeting alpha fetoprotein (AFP). This approach relies on a self-assembled heptamer fusion protein (A1-C4bpα), incorporating the dual functions of multivalent recognition and crosslinking aggregation amplification due to the presence of seven AFP-specific A1 nanobodies on the A1-C4bpα heptamer. Leveraging antibody-functionalized magnetic nanoparticles for target AFP capture and DLS signal output, the proposed heptamer-assisted DLS immunosensor offers high sensitivity, strong specificity, and ease of operation. Under the optimized conditions, the designed DLS immunosensor demonstrates excellent linear detection of AFP in the concentration range 0.06 ng mL-1 to 512 ng mL-1, with a detection limit of 15 pg mL-1. The selectivity, accuracy, precision, practicability, and reliability of this newly developed method were further validated through an assay of AFP levels in spiked and actual human serum samples. Thiswork introduces a novel approach for constructing ultrasensitive DLS immunosensors, easily extendable to the sensitive determination of other targets via simply replacing the nanobody sequence, holding great promise in various applications, particularly in disease diagnosis.

New design of microchip for in-situ DLS measurement of colloidal suspensions

Stability of dispersed materials remains an important question in a wide variety of fields such as cosmetics, catalysis, food or energy and the environment. As stability is directly linked to the size of the dispersed colloids it is essential to assess the size distribution of colloidal suspensions. Nowadays, microfluidic-based approaches generate increasing interest as they represent flexible and fast measurements allowing high throughput experimentations. However, characterization of colloidal dispersions is usually performed by dynamic light scattering (DLS), that requires static measurements as well as significant volumes, that are not compatible with on-line analysis and microfluidics. Moreover, due to flow-induced decorrelation terms, DLS measurements in microfluidic channels are only accurate at very low shear rates. This work aimed at developing an on-line microfluidic device for dispersed materials characterization using DLS. The main challenges of this project were i) to adapt the microfabrication of the PDMS device in order to combine microchannels of hundreds of microns with a milli-fluidic cavity to perform the DLS measurements, and ii) to downsize the DLS set up. A PDMS microchip, consisting in a millimeter cavity for DLS measurements in parallel with a microchannel, was designed to perform the measurement on the sample without stopping the suspension flow during the microfluidic experiment. The cavity geometry was then optimized thanks to numerical simulation to ensure a good sweep efficiency and to downscale as much as possible without impairing the DLS signal. By adapting the microfabrication process, a PDMS microfluidic chip was designed allowing the size measurements of successive suspensions containing 100 and 12 ​nm diameter particles. This work is a first step towards the implementation of a new technological building block for online microfluidic characterization. © 2017 Elsevier Inc. All rights reserved.

Rapid and sensitive determination of lactoferrin in milk powder by boronate affinity amplified dynamic light scattering immunosensor

Lactoferrin (Lf) is a bioactive multifunctional glycoprotein that belongs to the transferrin family. As an important nutritional fortifier, Lf is one of the star nutrients in milk powder. Thus, the rapid and accurate quantitative detection of Lf content in milk powder is crucial to the evaluation of its quality. However, most reported immunoassay methods for Lf quantitation suffer from cumbersome sample pretreatment or lengthy testing time and do not meet the requirements for rapid Lf detection in milk powder. Herein, we report a novel method combining dynamic light scattering (DLS) immunosensing and boronate affinity recognition for the rapid and sensitive detection of Lf. Owing to the selective binding between a boronate ligand and the cis diol, the developed DLS immunosensor quantitatively detected Lf with saving one antibody, greatly reducing the need for matching antibodies, which are needed in traditional sandwich immunoassay for biomolecular detection. In addition, the DLS immunosensor exhibited high selectivity and sensitivity to Lf, with a detection limit of 1 ng/mL within ∼15 min through coupling with magnetic nanoparticle-assisted immunomagnetic separation and DLS signal transduction.

Covalent organic framework-gold nanoparticle heterostructures amplified dynamic light scattering immunosensor for ultrasensitive detection of NT-proBNP in whole blood

Immunoassay has shown great potential in the detection of various disease protein biomarkers because of its high specificity. However, the sensitivity of conventional immunoassay is relatively limited due to its insufficient colorimetric signal transduction. Herein, we report a novel dynamic light scattering (DLS) immunosensing strategy for ultrasensitive detection of protein markers based on the covalent organic framework-gold nanoparticle (COF@AuNP) heterostructures combined with the gold growth. By taking advantage of the unique acid response characteristics of imine-linked COF, the COF@AuNP is designed as a signal amplifier to boost the release of AuNPs, and then the released AuNPs are enlarged by gold growth to amplify the DLS signal transduction and thus remarkably improve the sensitivity. Combined with the double antibody sandwich magnetic immunoassay, the COF@AuNP-amplified DLS immunosensor successfully achieve the highly sensitive quantitative detection of N-terminal brain-type natriuretic peptide. The limit of detection reaches up to 14 fg mL−1 with the total assay time less than 2 h. The linear detection range was from 0.32 to 1000 pg mL−1, and the coefficient of variation was below 12.5%. Briefly, this immunosensor is demonstrated with high selectivity, accuracy, reproducibility, and reliability in spiked and actual whole blood samples. The COF@AuNP-amplified DLS immunosensing strategy holds high promise as an alternative for detecting disease biomarkers.

Ultrasensitive dynamic light scattering immunosensing platform for NT-proBNP detection using boronate affinity amplification

Herein, we reported a new dynamic light scattering (DLS) immunosensing technology for the rapid and sensitive detection of glycoprotein N-terminal pro-brain natriuretic peptide (NT-proBNP). In this design, the boronate affinity recognition based on the interaction of boronic acid ligands and cis-diols was introduced to amplify the nanoparticle aggregation to enable highly sensitive DLS transduction, thereby lowering the limit of detection (LOD) of the methodology. After covalently coupling with antibodies, magnetic nanoparticles (MNPs) were employed as the nanoprobes to selectively capture trace amount of NT-proBNP from complex samples and facilitate DLS signal transduction. Meanwhile, silica nanoparticles modified with phenylboronic acid (SiO2@PBA) were designed as the crosslinking agent to bridge the aggregation of MNPs in the presence of target NT-proBNP. Owing to the multivalent and fast affinity recognition between NT-proBNP containing cis-diols and SiO2@PBA, the developed DLS immunosensor exhibited charming advantages over traditional immunoassays, including ultrahigh sensitivity with an LOD of 7.4 fg mL−1, fast response time (< 20 min), and small sample consumption (1 μL). The DLS immunosensor was further characterized with good selectivity, accuracy, precision, reproducibility, and practicability. Collectively, this work demonstrated the promising application of the designed boronate affinity amplified-DLS immunosensor for field or point-of-care testing of cis-diol-containing molecules.Graphical

Low-sample-consumption and ultrasensitive detection of procalcitonin by boronate affinity recognition-enhanced dynamic light scattering biosensor

Accurate determination of procalcitonin (PCT) is highly crucial in bacterial infection diagnosis. Many biosensors previously developed suffer from large sample consumption or lengthy waiting time, which raise difficulties for more vulnerable patients, such as infants, old people, and other critically ill patients. To address this dilemma, we present an innovative boronate affinity recognition (BAR)-enhanced dynamic light scattering (DLS) biosensor to achieve ultrasensitive PCT detection. In this biosensing system, monoclonal antibody-modified magnetic nanoparticles (MNP@mAb) are designed as probes to capture PCT from serum samples and generate DLS signal transduction. Polyvalent phenylboronic acid-labeled bovine serum albumin (BSA@PBA) is used as scaffold to aggregate MNP@mAb and PCT (MNP@mAb-PCT) complex because of the specific interaction of cis-diol-containing PCT with boronic acid ligands on the surface of BSA@PBA. The BAR-enhanced DLS biosensor shows ultrahigh sensitivity to PCT determination due to high binding affinity, with the limit of detection of 0.03 pg/mL. The total detection time of PCT in whole blood or serum is less than 15 min with small sample consumption (about 1 μL) due to the rapid magnetic separation and aggregation of MNP@mAb-PCT triggered by BSA@PBA. In addition, the proposed DLS biosensor exhibits a high specificity for PCT quantitative detection. Therefore, this work provides a promising and versatile strategy for extending DLS biosensor to rapid and ultrasensitive detection of trace PCT for broader patients and more urgent cases.

A Universal Boronate‐Affinity Crosslinking‐Amplified Dynamic Light Scattering Immunoassay for Point‐of‐Care Glycoprotein Detection

AbstractHerein, we report a universal boronate‐affinity crosslinking‐amplified dynamic light scattering (DLS) immunoassay for point‐of‐care (POC) glycoprotein detection in complex samples. This enhanced DLS immunoassay consists of two elements, i.e., antibody‐coated magnetic nanoparticles (MNP@mAb) for target capture and DLS signal transduction, and phenylboronic acid‐based boronate‐affinity materials as crosslinking amplifiers. Upon the addition of targets, glycoproteins are first captured by MNP@mAb and amplified by target‐induced crosslinking stemming from the selective binding between the boronic acid ligand and cis‐diol‐containing glycoprotein, thereby resulting in a remarkably increased DLS signal in the average nanoparticle size. Benefiting from the multivalent binding and fast boronate‐affinity reaction between glycoproteins and crosslinkers, the proposed immunosensing strategy has achieved the ultrasensitive and rapid quantitative assay of glycoproteins at the fM level within 15 min. Overall, this work provides a promising and versatile design strategy for extending the DLS technique to detect glycoproteins even in the field or at POC.

A Universal Boronate-Affinity Crosslinking-Amplified Dynamic Light Scattering Immunoassay for Point-of-Care Glycoprotein Detection.

Herein, we report a universal boronate-affinity crosslinking-amplified dynamic light scattering (DLS) immunoassay for point-of-care (POC) glycoprotein detection in complex samples. This enhanced DLS immunoassay consists of two elements, i.e., antibody-coated magnetic nanoparticles (MNP@mAb) for target capture and DLS signal transduction, and phenylboronic acid-based boronate-affinity materials as crosslinking amplifiers. Upon the addition of targets, glycoproteins are first captured by MNP@mAb and amplified by target-induced crosslinking stemming from the selective binding between the boronic acid ligand and cis-diol-containing glycoprotein, thereby resulting in a remarkably increased DLS signal in the average nanoparticle size. Benefiting from the multivalent binding and fast boronate-affinity reaction between glycoproteins and crosslinkers, the proposed immunosensing strategy has achieved the ultrasensitive and rapid quantitative assay of glycoproteins at the fM level within 15 min. Overall, this work provides a promising and versatile design strategy for extending the DLS technique to detect glycoproteins even in the field or at POC.

Direct competitive ELISA enhanced by dynamic light scattering for the ultrasensitive detection of aflatoxin B1 in corn samples

Compared with absorbance, scattering-based dynamic light scattering (DLS) signal has higher sensitivity because its light-scattering intensity is very sensitive to changes in size, thereby enhancing the sensitivity. Herein, we first developed a DLS-enhanced direct competitive enzyme-linked immunosorbent assay (DLS-dcELISA) for ultrasensitive detection of aflatoxin B1 (AFB1) in corn. By using hydroxyl radical-induced gold nanoparticle (AuNP) aggregation to amplify AuNP scattering signals, the developed DLS-dcELISA exhibited ultrahigh sensitivity for AFB1. The detection limit was 0.12 pg mL−1, which was 153- and 385-fold lower than those obtained using plasmonic and colorimetric dcELISA. In addition, the DLS-dcELISA exhibited excellent selectivity, high accuracy, and strong practicality. Overall, this work presented a simple and universal strategy for improving the sensitivity of traditional ELISA platform only by using the sensitive DLS signals. This technique can replace absorbance-based plasmonic or colored signals as immunoassay signal output for enhanced competitive detection of mycotoxins.

Size characterization and quantification of titanium dioxide nano- and microparticles-based products by Asymmetrical Flow Field-Flow Fractionation coupled to Dynamic Light Scattering and Inductively Coupled Plasma Mass Spectrometry

A procedure for the size characterization and quantification of titanium dioxide (TiO2) nano- and microparticles by Asymmetric Flow Field-Flow Fractionation (AF4) coupled to Dynamic Light Scattering (DLS) and Inductively Coupled Plasma Mass Spectrometry (ICP-MS) is described. Different strategies for size characterization with size standards and the use of the DLS signal for the estimation of hydrodynamic diameters are evaluated. The procedure has been applied to the characterization of TiO2 nanoparticles in photocatalytic products and crab sticks (surimis), where TiO2 is present as E171 food additive. Sizes in the range of 50–90 nm and 160–170 nm were estimated in the different photocatalytic products by AF4-DLS, in good agreement with the sizes predicted by calibration versus SiO2 and polystyrene standards. In surimis, sizes between 140 and 350 nm were estimated by AF4-DLS, similar to those reported in literature for E171 additive. These results were also compared to those obtained by single particle ICP-MS, which allowed the detection of a nano-sized fraction of TiO2 present in the four surimis analyzed. Titanium contents in one of the photocatalytic products determined by AF4-ICP-MS was 16.86 ± 2.54 mg g−1, whereas the alkaline extraction followed by AF4-ICP-MS allowed the determination of TiO2 content in four surimis at concentration levels in the range of the μg g−1 (from 3.14 ± 0.10 to 14.55 ± 1.46 μg Ti g−1), with channel recoveries above 85% in all cases. The method has been validated by comparison with the Ti content determined by ICP-OES after microwaved assisted acid digestion of all the samples. The methodology proposed allows the complete quantification of the (nano)particulate forms of titanium in complex matrices together with their size characterization.

Ultrasensitive dynamic light scattering based nanobiosensor for rapid anticancer drug screening

Improved chemical library screens for drug modulators of a target protein are vital to advance precision medicine. Here, we describe an ultrasensitive dynamic light scattering (DLS) based biosensor using gold nanoparticles as plasmon-enhanced sensing probes to rapidly screen anticancer compounds. The nanobiosensor which consists of DNA-conjugated dumbbell-shaped gold nanoprobes can detect protein activities in drug-treated cells. We have validated the utility of our nanosensor to screen for compounds known to reactivate mutated tumor suppressor protein p53 for DNA binding in complex cellular context, which leads to significant increases in the hydrodynamic size of the nanoprobes through DLS measurement. This unique nanoplasmonic biosensor not only significantly enhances the DLS signal for specific biomolecular binding, but also effectively suppresses the background noise from irrelevant entities, which greatly improves sensitivity and reduces consumption of both sample and probe. In addition, a competition assay designed to evaluate the relative DNA binding affinities of p53 using the same sensing probes allows a concomitant assessment of responsive p53 pathways downstream or cell fate. Most critically, this nanosensor enables direct interrogation of endogenous protein activities following drug treatment in live cells, allowing simultaneous on-target validation during phenotypic screens. This DLS-based nanobiosensor is broadly applicable to other DNA binding molecules and/or proteins, and holds great potential for high-throughput screening campaigns utilizing both conventional and fragment-based compound libraries in drug discovery.

Non-invasive Detection of Unique Molecular Signatures in Laser-Induced Retinal Injuries.

Unintentional laser exposure is an increasing concern in many operational environments. Determining whether a laser exposure event caused a retinal injury currently requires medical expertise and specialized equipment that are not always readily available. The purpose of this study is to test the feasibility of using dynamic light scattering (DLS) to non-invasively detect laser retinal injuries through interrogation of the vitreous humor (VH). Three grades of retinal laser lesions were studied: mild (minimally visible lesions), moderate (Grade II), and severe (Grade III). A pre-post-treatment design was used to collect DLS measurements in vivo at various time points, using a customized instrument. VH samples were analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS) and relative protein abundances were determined by spectral counting. DLS signal analysis revealed significant changes in particle diameter and intensity in laser-treated groups as compared with control. Differences in protein profile in the VH of the laser-treated eyes were noted when compared with control. These results suggest that laser injury to the retina induces upregulation of proteins that diffuse into the VH from the damaged tissue, which can be detected non-invasively using DLS.

Diffusivities of Ternary Mixtures of n-Alkanes with Dissolved Gases by Dynamic Light Scattering

Theoretical approaches suggest that dynamic light scattering (DLS) signals from low-molecular-weight ternary mixtures are governed by fluctuations in temperature as well as two individual contributions from fluctuations in concentration that are related to the eigenvalues of the Fick diffusion matrix. Until now, this could not be proven experimentally in a conclusive way. In the present study, a detailed analysis of DLS signals in ternary mixtures consisting of n-dodecane (n-C12H26) and n-octacosane (n-C28H58) with dissolved hydrogen (H2), carbon monoxide (CO), or water (H2O) as well as of n-C12H26 or n-C28H58 with dissolved H2 and CO is given for temperatures up to 523 K and pressures up to 4.1 MPa. Thermal diffusivities of pure n-C12H26 and n-C28H58 as well as thermal and mutual diffusivities of their binary mixtures being the basis for the ternary mixtures with dissolved gas were studied for comparison purposes. For the investigated ternary mixtures, three individual signals could be distinguished in the time-resolved analysis of scattered light intensity by using photon correlation spectroscopy (PCS). For the first time, it could be evidenced that these signals are clearly associated with hydrodynamic modes. In most cases, the fastest mode observable for ternary mixtures is associated with the thermal diffusivity. The two further modes obviously related to the molecular mass transport are observable on different time scales and comparable to the modes associated with the concentration fluctuations in the respective binary mixtures. Comparison of the experimental data with results from molecular dynamics simulations revealed very good agreement.

Simulation of Dynamic Light Scattering Signal Based on AR Model

According to the autocorrelation function (ACF) of the ultrafine particles, the auto-regressive (AR) model used for dynamic light scattering (DLS) signals simulation can be established. By the method, DLS signals for nanoparticles distribution are simulated respectively and every ACF of simulation signal performs a great agreement with its theory value. The simulation signals were respectively inversed and inversion errors are less than 5.25%. The particles size inversions of Photon Correlation Spectroscopy (PCS) are obtained by Morozov discrepancy principle. Experiments show that the method based on AR model is feasible in DLS signal simulation.

Simultaneous Determination of Thermal and Mutual Diffusivity of Binary Mixtures of n-Octacosane with Carbon Monoxide, Hydrogen, and Water by Dynamic Light Scattering

It is demonstrated that thermal and mutual diffusivities of binary mixtures of n-octacosane (n-C28H58) with carbon monoxide (CO), hydrogen (H2), and water (H2O) are simultaneously accessible by dynamic light scattering (DLS). As the light-scattering signals originating from thermal and concentration fluctuations appear in similar time scales, different data evaluation strategies were tested to achieve minimum uncertainties in the resulting transport properties. To test the agreement of the respective theoretical model with the DLS signals in the regression, an improved multifit procedure is introduced. With the selected data evaluation strategy, uncertainties of 4 to 15% and 4 to 30% in the thermal and mutual diffusivities, respectively, could be obtained for the binary mixtures. The mutual diffusivities for the mixtures measured at temperatures ranging from 398 to 523 K and pressures of 5 to 30 bar at saturation conditions are in good agreement with molecular dynamics simulations and data from the literature.

Mercuric ions induced aggregation of gold nanoparticles as investigated by localized surface plasmon resonance light scattering and dynamic light scattering techniques

With the development of nanosciences, both localized surface plasmon resonance light scattering (LSPR-LS) and dynamic light scattering (DLS) techniques have been widely used for quantitative purposes with high sensitivity. In this contribution, we make a comparison of the two light scattering techniques by employing gold nanoparticles (AuNPs) aggregation induced by mercuric ions. It was found that citrate-stabilized AuNPs got aggregated in aqueous medium in the presence of mercuric ions through a chelation process, resulting in greatly enhanced LSPR-LS signals and increased hydrodynamic diameter. The enhanced LSPR-LS intensity (ΔI) is proportional to the concentration of mercuric ions in the range of 0.4–2.5 μM following the linear regression equation of ΔI = −84.7+516.4c, with the correlation coefficient of 0.983 (n = 6) and the limit of determination (3σ) about 0.10 μM. On the other hand, the increased hydrodynamic diameter can be identified by the DLS signals only with a concentration of Hg2+ in the range of 1.0–2.5 μM, and a linear relationship between the average hydrodynamic diameters of the resulted aggregates and the concentration of Hg2+ can be expressed as d = −6.16 + 45.9c with the correlation coefficient of 0.994. In such case, LSPR-LS signals were further applied to the selective determination of mercuric ions in lake water samples with high sensitivity and simple operation.

New noninvasive index for evaluation of the vascular age of healthy and sick people

We conducted a study on 861 healthy and sick subjects and demonstrated that some calculated parameters based on measurement of the dynamic light scattering (DLS) signal from the finger correlate highly with chronological age ranging from 1.5 to 85 years old. Measurements of DLS signals were obtained during both occlusion and nonocclusion of blood flow in the finger. For the nonocclusion case we found that the low-frequency component of the DLS signal significantly correlates with the biological age while the high-frequency component of the DLS signal resembles the arterial pulse-wave and does correlate with age. However, the most prominent correlation between the DLS characteristics and age was noted with the stasis stage measurements. We propose that the observed age-related phenomena are caused by alterations in local blood viscosity and interactions of the endothelial cells with erythrocytes. Further, a new noninvasive index based on the age-related optical characteristics was introduced. This noninvasive index may be used as a research and diagnostic tool to examine the endothelial and thrombolytic properties of the vascular system.

Monitoring and Scoring Counter-Diffusion Protein Crystallization Experiments in Capillaries by in situ Dynamic Light Scattering

In this paper, we demonstrate the feasibility of using in situ Dynamic Light Scattering (DLS) to monitor counter-diffusion crystallization experiments in capillaries. Firstly, we have validated the quality of the DLS signal in thin capillaries, which is comparable to that obtained in standard quartz cuvettes. Then, we have carried out DLS measurements of a counter-diffusion crystallization experiment of glucose isomerase in capillaries of different diameters (0.1, 0.2 and 0.3 mm) in order to follow the temporal evolution of protein supersaturation. Finally, we have compared DLS data with optical recordings of the progression of the crystallization front and with a simulation model of counter-diffusion in 1D.

Simulation of Dynamic Light Scattering Signals of Ultrafine Particles

AbstractIn order to conveniently obtain dynamic light scattering (DLS) signal, a simulation method of DLS signal is proposed in this paper. This method regards the light intensity fluctuation of DLS as a stationary random process. According to autocorrelation function (ACF) of DLS signal, the stationary random process of DLS can be generated by auto‐regressive (AR) model. Signal simulations of several kinds of distribution particles and comparison of simulations and experiment prove that AR model can be used for DLS signal simulation. Moreover, by analyzing effect of simulation parameters on simulation precision, we obtained the relationship between simulation parameters and simulation precision. Finally, application of simulation signal verifies the effectiveness and convenience of this simulation method.

A non-invasive method for the assessment of hemostasis in vivo by using dynamic light scattering

We present a new non-invasive method for assessing hemostasis in vivo. This method is based on the analysis of the movement characteristics of red blood cells (RBCs) during blood stasis condition. Stasis is intermittently induced by occlusion of arterial blood flow at the finger root. We assumed that under zero flow conditions, RBC movement is driven mostly by Brownian motion, and we characterized the RBC movement by utilizing the dynamic light scattering (DLS) technique in vivo. We found that during the stasis the RBCs diffusion coefficient in plasma decreases. We speculate that the RBC diffusion coefficient is most strongly related to endothelial and hemostatic activity. This assumption is supported by our findings that RBC movement, being expressed through the characteristics of the measured DLS signal, is correlative to the biological age and also is related to the coagulation factors. This new method can serve as a new diagnostic and research tool for the assessment of hemostasis and vascular function.