Polystyrene Microparticles Research Articles

Detection of Influenza Virus by Agglutination of Microparticles Immobilized a Mixed Glycan Receptor Produced from Cells.

The hemagglutination inhibition (HAI) assay is one of the detection methods for influenza virus (IFV) under global influenza surveillance, which uses freshly prepared animal red blood cells (RBCs). Here, we demonstrate that a mixed glycan-modified polystyrene microparticle, which can be chemically prepared in advance, can replace animal RBCs in the HAI assay. A mixture of azide-conjugated glycans containing sialyl- and sulfated-lactose moieties was produced from Madin-Darby canine kidney (MDCK) cells, which are used for IFV isolation, and then immobilized on the surface of a polystyrene microparticle using click chemistry. Human HA and IFV were detected with high sensitivity when using the mixed glycan-immobilized particle.

Direct Quantification of Proteins Modified on a Polystyrene Microparticle Surface Based on ζ Potential Change.

The zeta potential (ζ) of a particle is a surface charge density (σ)-dependent parameter. If a change in σ can be induced by surface modification, the number of molecules modified on the particle can be detected as a measurable change in ζ. In this study, we demonstrate protein detection at zmol or pM levels (bovine serum albumin (BSA), myoglobin (Mb), and lysozyme (Lyz)) on carboxy-functionalized polystyrene (PS) microparticles using the ζ change. Protein modification of the PS particles changes σ because the negatively charged carboxy group is used for protein binding, and proteins also have charged amino acids. The pH dependence of ζ for the protein-modified particles at 4 < pH < 10 is well-explained using the acid dissociation of the acidic and basic amino acids and the Gouy-Chapman-Stern model. An increase in the binding number of proteins per single PS particle (npro/PS) leads to a decrease in ζ, which is consistent with the results estimated by the proposed model. The detection limits of nBSA/PS, nMb/PS, and nLyz/PS are 1.17 × 104, 1.22 × 104, and 1.20 × 104 at pH 8.52, respectively, which means that the concentration-based detection limits are 722, 376, and 371 pM, respectively. We expect that the present method will be a strategy for the detection of molecules on particles.

Immuno-Modulatory Effects of Microparticles Formulated from Degradable Polystyrene Analogue.

Environmental accumulation of non-degradable polystyrene (PS) microparticles from plastic waste poses potential adverse impact on marine life and human health. Herein, microparticles from a degradable PS analogue (dePS) are formulated and their immuno-modulatory characteristics are comprehensively evaluated. Both dePS copolymer and microparticles are chemically degradable under accelerated hydrolytic condition. Invitro studies show that dePS microparticles are non-toxic to three immortalized cell lines. While dePS microparticles do not induce macrophage polarization invitro, dePS microparticles induce invivo upregulation of both pro-inflammatory and anti-inflammatory biomarkers in immuno-competent mice, suggesting the coexistence of mixed phenotypes of macrophages in the host immune response to these microparticles. Interestingly, on day7 following subcutaneous in mice, dePS microparticles induce a lower level of several immuno-modulatory biomarkers (matrix metallo-proteinases (MMPs), tumor necrosis factor (TNF-α), and arginase activity) compared to that of reference poly(lactic-co-glycolic acid) microparticles. Remarkably, compared to PS microparticles, dePS microparticles exhibit similar invitro and invivo bioactivity while acquiring additional chemical degradability. Overall, this study gains new insights into the host immune response to dePS microparticles and suggests that this dePS analogue might be explored as an alternative material choice for biomedical and consumer care applications.

Rapid Detection of Gram-Positive and -Negative Bacteria in Water Samples Using Mannan-Binding Lectin-Based Visual Biosensor.

Waterborne bacterial infection is a health threat worldwide, making accurate and timely bacteria detection crucial to prevent waterborne disease outbreaks. Inspired by the intrinsic capability of mannan-binding lectin (MBL) in recognizing the pathogen-associated molecular patterns (PAMPs), a visual biosensor is developed here for the on-site detection of both Gram-positive and -negative bacteria. The biosensor was synthesized by immobilization of the MBL protein onto the blue carboxyl-functionalized polystyrene microparticles (PSM), which is then used in a two-step assay to detect bacterial cells in water samples. The first step involved a 20 min incubation following the MBL-PSM and calcium chloride solution addition to the samples. The second step was to add ethanol to the resultant blue mixture and observe the color change with the naked eye after 15 min. The biosensor had a binary (all-or-none) response, which in the presence of bacterial cells kept its blue color, while in their absence the color changed from blue to colorless. Testing the water samples spiked with four Gram-negative bacteria including Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa and two Gram-positive bacteria of Enterococcus faecalis and Staphylococcus aureus showed that the biosensor could detect all tested bacteria with a concentration as low as 101.5 CFU/ml. The performance of biosensor using the water samples from a water treatment plant also confirmed its capability to detect the pathogens in real-life water samples without the need for instrumentation.

Analysis of thermophoresis for separation of polystyrene microparticles in microfluidic devices

We analysed the thermodiffusion phenomenon both numerically and experimentally for size-based separation of polystyrene microparticles. For model validation, we followed previously published numerical studies using ANSYS Fluent 2020 R2 software. For our experimental analysis, we defined a new microchannel geometry that would separate at least two groups of particles (5 and 20 µm). We analysed the trajectory of the microparticles in the central channel of the microdevice under the following conditions: without a temperature gradient, with application of a thermal gradient parallel to the gravitational field (cooling from the bottom or top part), and generation of a temperature gradient perpendicular to the direction of the gravity force. Numerical and experimental results for these geometry and boundary conditions demonstrated that, under terrestrial conditions, 5 µm and larger microsized polystyrene particles cannot be separated by thermophoresis in flow because of the gravity force.

Fluid dynamics and cell-bound Psl polysaccharide allows microplastic capture, aggregation and subsequent sedimentation by Pseudomonas aeruginosa in water.

SummaryDecades after incorporating plastics into consumer markets, research shows that these polymers have spread worldwide. Fragmentation of large debris leads to smaller particles, collectively called microplastics (MPs), which have become ubiquitous in aquatic environments. A fundamental aspect of understanding the implications of MP contamination on ecosystems is resolving the complex interactions of these artificial substrates with microbial cells. Using polystyrene microparticles as model polymers, we conducted an exploratory study where these interactions are quantitatively analyzed using an in vitro system consisting of single‐bacterial species capturing and aggregating MPs in water. Here we show that the production of Psl exopolysaccharide by Pseudomonas aeruginosa (PA) does not alter MPs colloidal stability but plays a key role in microspheres adhesion to the cell surface. Further aggregation of MPs by PA cells depends on bacterial mobility and the presence of sufficient flow to prevent rapid sedimentation of early MP‐PA assembles. Surprisingly, cells in MP‐PA aggregates are not in a sessile state despite the production of Psl, enhancing the motility of the aggregates by an order of magnitude relative to passive diffusion. The generated data could inform the creation of predictive models that accurately describe the dynamics and influence of bacterial growth on plastics debris.

Antibody-functionalized aluminum oxide-coated particles targeting neutrophil receptors in a multifrequency microfluidic impedance cytometer.

Personalized diagnostics of infectious diseases require monitoring disease progression due to their ever-changing physiological conditions and the multi-faceted organ system mechanisms involved in disease pathogenesis. In such instances, the recommended clinical strategies involve multiplexing data collection from critical biomarkers related to a patient's conditions along with longitudinal frequent patient monitoring. Numerous detection technologies exist both in research and commercial settings to monitor these conditions, however, they fail to provide biomarker multiplexing ability with design and data processing simplicity. For a recently conceived multiplexing biomarker modality, this work demonstrates the use of electrically sensitive microparticles targeting and identifying membrane receptors on leukocytes using a single detection source, with a high potential for multiplexing greater than any existing impedance-based single-detection scheme. Here, polystyrene microparticles are coated with varying thicknesses of metal oxides, which generate quantifiable impedance shifts when exposed to multifrequency electric fields depending on the metal oxide thickness. Using multifrequency impedance cytometry, these particles can be measured and differentiated rapidly across one coplanar electrode scheme. After surface-functionalizing particles with antibodies targeting CD11b and CD66b receptors, the particles are combined with isolated neutrophils to measure receptor expression. A combination of data analysis techniques including multivariate analysis, supervised machine learning, and unsupervised machine learning was able to accurately differentiate samples with up to 91% accuracy. This proof-of-concept study demonstrates the potential for these oxide-coated particles for enumerating specific leukocytes enabling multiplexing. Further, additional coating thicknesses or different metal oxide materials can enable a compendium of multiplexing targeting resource to be used to develop a high-multiplexing sensor for targeting membrane receptor expression.

Experimental and theoretical studies of cross-stream migration of non-spherical particles in a quadratic flow of a viscoelastic fluid.

We experimentally investigate the cross-stream migration of spherical, prolate, and oblate particles in a circular tube flow of a weakly viscoelastic fluid (De = O(10-2)) with negligible inertia (Re ≈ 0). From our previous theoretical studies, we developed mathematical models based on a second order fluid (i.e., retarded expansion for De ≪ 1) to characterize the migration trajectory of the particles in the absence of wall effects. The theory shows that the particle migration speed is proportional to the length the particle spans in the shear gradient direction (Lsg), and furthermore quantifies how particle shape alters the migration timescale. For particles with identical volume, spherical particles show the fastest migration speed among all the particles. The distinctive orientation behavior of prolate and oblate spheroids leads to a faster migration speed for an oblate particle compared to a prolate particle of the same aspect ratio. In this work, we verify our theory with microfluidic flow experiments using a model suspension of polystyrene (PS) micro-particles in a density-matched polyvinylpyrrolidone (PVP) solution (a Boger fluid). The experimental results show good qualitative and quantitative agreement with the theoretically predicted particle migration speed, indicating that the theory is able to provide reasonable predictions for real microfluidic systems.

DNA-based artificial dendritic cells for in situ cytotoxic T cell stimulation and immunotherapy

In immunotherapy, ex vivo stimulation of T cells requires significant resources and effort. Here, we report artificial dendritic cell-mimicking DNA microflowers (DM) for programming T cell stimulation in situ. To mimic dendritic cells, DNA-based artificial dendritic microflowers were constructed, surface-coated with polydopamine, and further modified with anti-CD3 and anti-CD28 antibodies to yield antibody-modified DM (DM-A). The porous structure of DM-A allowed entrapment of the T cell-stimulating cytokine, ineterleukin-2, yielding interleukin-2-loaded DM-A (DM-AI). For comparison, polystyrene microparticles coated with polydopamine and modified with anti-CD3 and anti-CD28 antibodies (PS-A) were used. Compared to PS-A, DM-AI showed significantly greater contact with T cell surfaces. DM-AI provided the highest ex vivo expansion of cytotoxic T cells. Local injection of DM-AI to tumor tissues induced the recruitment of T cells and expansion of cytotoxic T cells in tumor microenvironments. Unlike the other groups, model animals injected with DM-AI did not exhibit growth of primary tumors. Treatment of mice with DM-AI also protected against growth of a rechallenged distant tumor, and thus prevented tumor recurrence in this model. DM-AI has great potential for programmed stimulation of CD8+ T cells. This concept could be broadly extended for the programming of specific T cell stimulation profiles.

Visual Quantitation of Copper Ions Based on a Microfluidic Particle Dam Reflecting the Cu(II)-Catalyzed Oxidative Damage of DNA.

Due to the use of copper water pipes and the discharge of industrial wastewater, contamination of copper ions in drinking water has become a severe hazard globally. To routinely check water safety on a daily basis, easy-to-use platforms for quantitative analysis of trace amounts of copper ions (Cu2+) in drinking water is needed. Here, we report microfluidic particle accumulation integrated with a Cu(II)-catalyzed Fenton reaction for visual and quantitative copper ion detection. Microparticles (MMPs) and polystyrene microparticles (PMPs) are connected via a single strand DNA, MB155. However, when Cu2+ is present, MB155 is cleaved by hydroxyl free radicals (•OH) produced from Cu2+/hydrogen peroxide (H2O2) Fenton reactions, causing an increased amount of free PMPs. To visually count them, the particle solution is loaded onto a microfluidic chip where free MMPs and MMPs–MB155–PMPs can be collected by the magnetic separator, while the free PMPs continue flowing until being accumulated at the particle dam. The results showed a good linear relationship between the trapping length of PMP accumulation and the Cu2+ concentration from 0 to 300 nM. A limit of detection (LOD) of 70.1 nM was achieved, which is approximately 449 times lower than the 2 × 103 μg·L−1 (~31.5 μM) required by the World Health Organization (WHO). Moreover, the results showed high selectivity and good tolerance to pH and hardness, indicating compatibility for detection in tap water, suggesting a potential platform for the routine monitoring of copper contamination in drinking water.

Supposedly identical microplastic particles substantially differ in their material properties influencing particle-cell interactions and cellular responses

Microplastics and its putative adverse effects on environmental and human health increasingly gain scientific and public attention. Systematic studies on the effects of microplastics are currently hampered by using rather poorly characterised particles, leading to contradictory results for the same particle type. Here, surface properties and chemical composition of two commercially available nominally identical polystyrene microparticles, frequently used in effect studies, were characterised. We show distinct differences in monomer content, ζ-potentials and surface charge densities. Cells exposed to particles showing a lower ζ-potential and a higher monomer content displayed a higher number of particle-cell-interactions and consequently a decrease in cell metabolism and proliferation, especially at higher particle concentrations. Our study emphasises that no general statements can be made about the effects of microplastics, not even for the same polymer type in the same size class, unless the physicochemical properties are well characterised.

Colloidal Photonic Crystals of Reusable Hydrogel Microparticles for Sensor and Laser Applications.

Although a wide variety of techniques have been developed to date for the fabrication of high-quality colloidal photonic crystals (CPCs) using monodisperse silica and polystyrene microparticles, poly(N-isopropylacrylamide) (PNIPA) hydrogel microparticles have rarely been utilized for the preparation of active CPCs despite the intriguing feature of temperature-responsive volume changes. This report describes the promising potential abilities of PNIPA hydrogel microparticles for sensor and laser applications. Monodisperse PNIPA hydrogel microparticles were synthesized by emulsion polymerization, and the microparticle diameter was finely controlled by adjusting the surfactant concentration. Such hydrogel microparticles spontaneously formed uniform CPCs with visible Bragg reflection even in fluid suspensions. The addition of small amounts of ionic substances into the centrifuged and deionized CPC suspensions enabled the on-demand color switching between Bragg reflection and white turbidity with temperature, leading to temperature- and ion-sensing applications. Moreover, our expanding experiments successfully demonstrated the optically excited laser action with a single and narrow peak from CPC suspensions with light-emitting dyes by the photonic band gap effect. After the light-emitting dyes were simply removed from the CPC suspensions by centrifugation, the purified PNIPA hydrogel microparticles were permanently reusable as the CPC laser microcavities to generate the laser action at other wavelengths using different dyes. This study contributes the circular economy concept using reusable hydrogel microparticles for the realization of a sustainable society.

Optimization of Photooxygenation of Dihydroartemisinic Acid in a Continuous Micro-reactor utilizing Physically Immobilized Methylene Blue on Polystyrene Micro-particles

Extensive studies reported upon the application of microfluidic devices for photooxygenation reactions. Only a few researches, however, have studied the optimization of experimentally controllable parameters towards achieving the best reaction conversion. Accordingly, in the current study, the optimum values of operating parameters towards achieving the highest dihydroartemisinic acid (DHAA) conversion were determined through the response surface methodology (RSM) technique. In order to do that, first, heterogenized micro-photosensitizers were synthesized during a super facile procedure not requiring any chemical reaction or modifications. Then, these micro-photosensitizers were utilized to perform continuous photooxygenation of the DHAA by means of a glass micro-reactor at room temperature and atmospheric pressure. Finally, the impacts of experimental parameters such as the flow ratios of solution to oxygen as well as DHAA and micro-photosensitizers’ concentrations upon the DHAA conversions were examined by using Design of Experiments (DOE) software in order to optimize and statistically model the DHAA conversion. The optimum value for this conversion was predicted by the RSM technique to be 70 % which was in a very good agreement with that of the experimental value of 69 % obtained in this study. The optimal values for the flow rates ratio, DHAA initial concentration, and micro-photosensitizer’s concentration were obtained to be 0.5, 0.5 mg/ml, as well as 1.5 mg/ml, respectively.

Label-free identification of microplastics in human cells: dark-field microscopy and deep learning study.

The development of an automatic method of identifying microplastic particles within live cells and organisms is crucial for high-throughput analysis of their biodistribution in toxicity studies. State-of-the-art technique in the data analysis tasks is the application of deep learning algorithms. Here, we propose the approach of polystyrene microparticle classification differing only in pigmentation using enhanced dark-field microscopy and a residual neural network (ResNet). The dataset consisting of 11,528 particle images has been collected to train and evaluate the neural network model. Human skin fibroblasts treated with microplastics were used as a model to study the ability of ResNet for classifying particles in a realistic biological experiment. As a result, the accuracy of the obtained classification algorithm achieved up to 93% in cell samples, indicating that the technique proposed will be a potent alternative to time-consuming spectral-based methods in microplastic toxicity research.

Assessment of the Effects of Environmental Concentrations of Microplastics on the Aquatic Snail Potamopyrgus antipodarum

Microplastics are ubiquitous in aquatic ecosystems. They can be found at the surface, in the water column, and in sediments. Multiple negative impacts of microplastics on aquatic organisms have been reported, with most studies focusing on marine ecosystems. However, the effects of microplastics on freshwater ecosystems have been less studied, with a few studies focusing on benthic invertebrates. In this study, we exposed the New Zealand mud snail Potamopyrgus antipodarum (Gray, 1843) to an environmental range of concentrations of polystyrene microparticles (size range from 0.01 to 514 µm at 100, 500, and 1000 mg microplastics/kg dry weight (dw) of sediment) and two supra-environmental concentrations (2000 and 4000 mg/kg dw sediment). The impacts of the exposure to microplastics on mortality, behavior, and reproduction were assessed at long-term exposure (31 days). Mortality and reproduction were not significantly affected by microplastics. On the contrary, most of the microplastic treatments altered the behavior, causing a significant increase in reaction time compared with controls (0 mg microplastics/kg dw sediment). The highest concentration (4000 mg/kg) did not have an impact on the reaction time over the experimental period compared with controls. To our knowledge, this study is the first to assess the effects of microplastics on the behavior of the aquatic snail P. antipodarum. Our results showed that at environmental concentrations, the behavior of P. antipodarum was the most sensitive variable to the adverse effects of polystyrene microplastics.

Optical Oxygen Sensor Patch Printed with Polystyrene Microparticles-based Ink on Flexible Substrate.

Optical oxygen sensors based on photoluminescence quenching have gained increasing attention as a superior method for continuous monitoring of oxygen in a growing number of applications. A simple and low-cost fabrication technique was developed to produce sensor arrays capable of two-dimensional oxygen tension measurement. Sensor patches were printed on polyvinylidene chloride film using an oxygen-sensitive ink cocktail, prepared by immobilizing Pt(II) mesotetra(pentafluorophenyl)porphine (PtTFPP) in monodispersed polystyrene microparticles. The dispersion media of the ink cocktail, high molecular weight polyvinyl pyrrolidone suspended in 50% ethanol (v/v in water), allowed adhesion promotion and compatibility with most common polymeric substrates. Ink phosphorescence intensity was found to vary primarily with fluorophore concentration and to a lesser extent with polystyrene particle size. The sensor performance was investigated as a function of oxygen concentrations employing two different techniques: a multi-frequency phase fluorometer and smart phone-based image acquisition. The printed sensor patch showed fast and repetitive response over 0-21% oxygen concentrations with high linearity (with R2 >0.99) in a Stern-Volmer plot, and sensitivity of I0/I21 >1.55. The optical sensor response on a surface was investigated further using two-dimensional images which were captured and analyzed under different oxygen environment. Printed sensor patch along with imaging read-out technique make an ideal platform for early detection of surface wounds associated with tissue oxygen.

Fabrication of Ring-Shaped Deposits of Polystyrene Microparticles Driven by Thermocapillary Mechanism.

Fabrication of ring-shaped deposits of microparticles on solid surfaces with the desired length scales and morphology of particle arrangements is of great importance when developing modern optical and electronic resonators, chemical sensors, touch screens, field-emission displays, porous materials, and coatings with various functional properties. However, the controlled formation of ring-shaped patterns scaling from a few millimeters up to centimeters with simultaneous control of particle arrangement at the microscale is one of the most challenging problems in advanced materials science and technology. Here, we report a fabrication approach for ring-shaped structures of microparticles on a glass surface that relied on a local thermal impact produced by the subsurface heater and heat sink. Thermocapillary convection in the liquid covering microparticles in combination with evaporative lithography is responsible for the particle transport and the assembling into the ring-shaped patterns. An advantageous feature of this approach is based on the control of thermocapillary flow direction, achieved by changing the sign of the temperature gradient in the liquid, switching between heating and cooling modes. That allows for changing the particle transfer direction to create the ring-shaped deposits and dynamically tune their size and density distribution. We have studied the influence of the power applied to the heat source/sink and the duration of the applied thermal field on the rate of the ring fabrication, the sizes of the ring and the profile of the particle distribution in the ring. The proposed method is flexible to control simultaneously the centimeter scale and microscale processes of transfer and arrangements of particles and can be applied to the fabrication of ring structures of particles of different nature and shape.

Cooperative Optical Trapping of Polystyrene Microparticle and Protein Forming a Submillimeter Linear Assembly of Microparticle

Cooperative Optical Trapping of Polystyrene Microparticle and Protein Forming a Submillimeter Linear Assembly of Microparticle

Time-domain signal averaging to improve microparticles detection and enumeration accuracy in a microfluidic impedance cytometer.

Microfluidic impedance cytometry is a powerful system to measure micro and nano-sized particles and is routinely used in point-of-care disease diagnostics and other biomedical applications. However, small objects near a sensor's detection limit are plagued with relatively significant background noise and are difficult to identify for every case. While many data processing techniques can be utilized to reduce noise and improve signal quality, frequently they are still inadequate to push sensor detection limits. Here, we report the first demonstration of a novel signal averaging algorithm effective in noise reduction of microfluidic impedance cytometry data, improving enumeration accuracy, and reducing detection limits. Our device uses a 22 µm tall × 100 µm wide (with 30 µm wide focused aperture) microchannel and gold coplanar microelectrodes that generate an electric field, recording bipolar pulses from polystyrene microparticles flowing through the channel. In addition to outlining a modified moving signal averaging technique theoretically and with a model data set, we also performed a compendium of characterization experiments including variations in flow rate, input voltage, and particle size. Multivariate metrics from each experiment are compared including signal amplitude, pulse width, background noise, and signal-to-noise ratio (SNR). Incorporating our technique resulted in improved SNR and counting accuracy across all experiments conducted, and the limit of detection improved from 5 to 1 µm particles without modifying microchannel dimensions. Succeeding this, we envision implementing our modified moving average technique to develop next-generation microfluidic impedance cytometry devices with an expanded dynamic range and improved enumeration accuracy. This can be exceedingly useful for many biomedical applications, such as infectious disease diagnostics where devices may enumerate larger-scale immune cells alongside sub-micron bacterium in the same sample.

Reconfigurable millimeter-range optical binding of dielectric microparticles in hollow-core photonic crystal fiber.

Optical binding of microparticles offers a versatile playground for investigating the optomechanics of levitated multi-particle systems. We report millimeter-range optical binding of polystyrene microparticles in hollow-core photonic crystal fiber. The first particle scatters the incident LP01 mode into several LP0n modes, creating a beat pattern that exerts a position-dependent force on the second particle. Particle binding results from the interplay of the forces created by counterpropagating beams. A femtosecond trapping laser is used so that group velocity walk-off eliminates disturbance caused by higher order modes accidentally excited at the fiber input. The inter-particle distance can be optically switched over 2 orders of magnitude (from 42 µm to 3 mm), and the bound particle pairs can be translated along the fiber by unbalancing the powers in the counterpropagating trapping beams. The frequency response of a bound particle pair is investigated at low gas pressure by driving with an intensity-modulated control beam. The system offers new degrees of freedom for manipulating the dynamics and configurations of optically levitated microparticle arrays.