Polystyrene Nanoparticles Research Articles

Photonic Microbead Array Digital Time-Resolved Fluorescence Ultrasensitive Platform for Simultaneous Detection of Multiple Mycotoxins.

Limitations in the sensitivity, linear detection range, and cross-reaction of lateral flow immunoassays mainly hamper their application in rapid screening for multiple targets. In this work, we designed a new time-resolved fluorescence immunoassay (TRFIA) platform to overcome these limitations. This platform uses europium chelate polystyrene (PS@Eu) nanoparticles conjugated with monoclonal antibodies to sense multiple mycotoxins. We employed a competitive TRFIA protocol in which the conjugated PS@Eu was used on the surfaces of photonic microbead arrays (PMAs). The TRFIA signal of PMAs on the pad was recorded with the digital time-resolved fluorescence reader. The developed TRFIA shows wide detection linear ranges (0.01-1000 ng/mL for DON, 0.1-100 ng/mL for OTA, and 0.01-100 ng/mL for AFB1), low limits of detection (LODs) (7.9 pg/mL for DON, 18 pg/mL for OTA, and 7.7 pg/mL for AFB1), good specificity, good recovery ratios (76.68-117.26%), and good reproducibility in grain samples. The simulated fluorescence enhancement effect of PMA indicated that the electric field distribution on the surface of PS@Eu on PMA is twice higher than that on the surface of PS@Eu. The new TRFIA for three kinds of mycotoxins was 1000-fold more sensitive than the classical TRFIA, and it has great potential application in rapid screening for multiple targets.

Detection of Tn-antigen in breast and prostate cancer models by VVL-labeled red dye-doped nanoparticles.

Aim: Fluorescence detection of breast and prostate cancer cells expressing Tn-antigen, a tumor marker, with Vicia villosa lectin (VVL)-labeled nanoparticles.Materials & methods: Breast and prostate cancer cells engineered to express high levels of Tn-antigen and non-engineered controls were incubated with VVL-labeled or unlabeled red dye-doped silica-coated polystyrene nanoparticles. The binding to cells was studied with flow cytometry, confocal microscopy, and electron microscopy.Results: Flow cytometry showed that the binding of VVL-labeled nanoparticles was significantly higher to Tn-antigen-expressing cancer cells than controls. Confocal microscopy demonstrated that particles bound to the cell surface. According to the correlative light and electron microscopy the particles bound mostly as aggregates.Conclusion: VVL-labeled nanoparticles could provide a new tool for the detection of Tn-antigen-expressing breast and prostate cancer cells.

Accumulation of nanoplastics by wheat seedling roots: Both passive and energy-consuming processes

Nanoplastics can transfer from the environment to plants and potentially harm organisms. However, the mechanisms on how crop root systems absorb and transport nanoplastics are still unclear. Here, original and fluorescent labeled polystyrene and polyvinyl chloride nanoparticles (PS-NPs, PVC-NPs; 30 nm; 10 mg L−1) were employed to study the distribution and internalization pathways in wheat seedling roots. In the study, nanoplastics accumulated more in the root tip and surface, with PVC-NPs more prevalent than PS-NPs. After being treated with inhibitors (Na3VO4, chlorpromazine and amiloride), the nanoplastics mean fluorescence intensities were reduced by 4.0–51.1 %. During the uptake, both passive and energy-consuming pathways occurred. For the energy-consuming uptake pathway, macropinocytosis contributed more to cytoplasm than clathrin-mediated endocytosis. H+ influx was observed during nanoplastic transport into the cytoplasm, and the reduction in plasma membrane ATPase activity led to a decrease in nanoplastic internalization. These results elucidate the pathways of nanoplastics absorption and transport in wheat roots, provide crucial evidence for assessing nanoplastics' ecological risks and support the development of technologies to block nanoplastics absorption by crop roots, ensuring agricultural and ecosystem safety.

Impact of dissolved organic matter characteristics and inorganic species on the stability and removal by coagulation of nanoplastics in aqueous media

The aggregation of rough, raspberry-type polystyrene nanoparticles (PS-NPs) was investigated in the presence of six hydrophobic and hydrophilic dissolved organic matter (DOM) isolates and biopolymers (effluent OM) in NaCl and CaCl2 solutions using time-resolved dynamic light scattering. Results showed that the stability of PS-NPs mainly depends on OM characteristics and ionic composition. Due to cation bridging, the aggregation rate of PS-NPs in Ca2+-containing solutions was significantly higher than at similar Na+-ionic strength. Biopolymers rich in protein and carbohydrate moieties showed higher affinity to the surface of PS-NPs than the other DOM isolates in the absence of both Ca2+ and Na+. Overall, the stability of PS-NPs followed the order of biopolymers > hydrophobic isolates > hydrophilic isolates in the presence of Na+ and biopolymers > hydrophilic isolates > hydrophobic isolates in Ca2+-containing solutions. In the presence of high MW structures (biopolymers), PS-NPs aggregation in both NaCl and CaCl2 solutions was attributed to steric repulsive forces. The impact of hydrophilic and hydrophobic isolates on PS-NPs aggregation highly relied on the ionic composition. Coagulation was an effective pretreatment for PS-NPs removal. Using inductively coupled plasma-mass spectrometry, higher removals were recorded with Al2(SO4)3 in the absence of DOM, while PACl more efficiently coagulated PS-NPs in the presence of DOM isolates.

Lack of effects of polystyrene micro- and nanoplastics on activity and expression of human drug transporters

Micro- and nanoplastics (MPs/NPs) constitute emerging and widely-distributed environmental contaminants to which humans are highly exposed. They possibly represent a threat for human health. In order to identify cellular/molecular targets for these plastic particles, we have analysed the effects of exposure to manufactured polystyrene (PS) MPs and NPs on in vitro activity and expression of human membrane drug transporters, known to interact with chemical pollutants. PS MPs and NPs, used at various concentrations (1, 10 or 100µg/mL), failed to inhibit efflux activities of the ATP-binding cassette (ABC) transporters P-glycoprotein, MRPs and BCRP in ABC transporter-expressing cells. Furthermore, PS particles did not impair the transport of P-glycoprotein or BCRP substrates across intestinal Caco-2 cell monolayers. Uptake activities of solute carriers (SLCs) such as OCT1 and OCT2 (handling organic cations) or OATP1B1, OATP1B3, OATP2B1, OAT1 and OAT3 (handling organic anions) were additionally not altered by PS MPs/NPs in HEK-293 cells overexpressing these SLCs. mRNA expression of ABC transporters and of the SLCs OCT1 and OATP2B1 in Caco-2 cells and human hepatic HepaRG cells were finally not impaired by a 48-h exposure to MPs/NPs. Altogether, these data indicate that human drug transporters are unlikely to be direct and univocal targets for synthetic PS MPs/NPs.

Functionalized europium infused-polystyrene nanoparticles for bacteria labeling for microscopic imaging

Luminescence microscopic imaging has become an indispensable tool in diagnostics and therapeutics. Lanthanide luminescent complexes (LLCs) have proven to be invaluable in this endeavor due to certain attractive characteristics such as long decay times, large Richardson shifts and narrow emission bands. However, the insolubility of these complexes and the negative effects of the aqueous environment on their photophysical properties still restrict their biological applications. By embedding luminescent Eu3+ complexes into polystyrene nanoparticles (NPs), these complexes can not only maintain their photophysical properties, but even enhance them. This leads to exceptionally high quantum yields (> 90%) and decay times ( >800 μs). In addition, the surfaces of the NPs described here are covered with primary amine groups, which may be exploited for various interesting bioconjugation chemistries. In the present application, they were modified with N-hydroxysuccinimide (NHS) esters and then coupled either electrostatically or covalently to both, gram positive and negative bacteria, labeling them for luminescence microscopic imaging. This, to the best of our knowledge, is the first report of polymer NPs infused with LLCs for bacteria labeling.

Preparation, characterization and thermal properties of the PS+Si based polymer nanocomposites

Polymer nanocomposites based on polystyrene and polycrystalline Si nanoparticles have been successfully synthesized using the solution-casting method. The scanning electron microscope and x-ray diffraction methods show that the Si nanoparticle size in the obtained nanocomposite was up to 50 nm. The thermostability of the obtained nanocomposite was studied depending on the concentration of Si nanoparticles in the PS matrix. It was found that Si nanoparticles that are distributed homogeneously in the polymer matrix act as a thermal barrier. By adding 3% Si nanoparticles into polystyrene polymer, the thermal degradation temperature of PS increases by 20°, and the decomposition rate reduces from −42,460 mg/min to −35,099 mg/min. In addition, it was shown that Si nanoparticles do not affect the PS glass transition temperature, which explains that the porous Si does not reduce the free volume for the polymer molecules to move. The polymer macromolecules just are adsorbed on the porous silicon surface, which partially limits their mobility.

A multi-analytical approach to evaluate the removal efficiency of polystyrene nanoparticles in water treatment processes

The removal of nanoplastics (NP) from water using various treatment processes has gained significant attention recently. This study comprehensively characterizes the degradation of polystyrene nanoparticles (concentration: 200 ppm, diameter: 140 nm) through UVC irradiation. For the first time, we compared four analytical methods to monitor removal efficiency: Py-GCMS, UV–Visible spectroscopy, TOC, and Turbidity. Additionally, DLS, TEM, and SEC were used to understand changes in particle size, morphology, and molecular weight. Results showed that Py-GCMS overestimated the removal rate by a factor of 2 compared to Turbidity and UV–Visible measurements, which were in agreement. Furthermore, after 200 h of irradiation, the styrene signal disappears from the pyrogram, although the mineralization rate reaches only 50%, as determined by total organic carbon (TOC) analysis. The particle size decreased slowly, reaching 100 nm after 150 h, while a significant decrease in molecular weight indicated high chain-scission. These findings emphasize the importance of a multi-analytical approach to accurately assess NP removal efficiency and understand degradation mechanisms.

Activated hedgehog and insulin ligands by decreased transcriptional factor DAF-16 mediate transgenerational nanoplastic toxicity in Caenorhabditis elegans

In Caenorhabditis elegans, transcriptional factor DAF-16 in insulin signaling pathway played important role in regulating transgenerational nanoplastic toxicity. Activation of insulin signals mediated transgenerational toxicity of polystyrene nanoparticle (PS-NP) by inhibiting DAF-16. Among identified germline ligands, expression of wrt-3 encoding hedgehog ligand was increased by RNAi of daf-16 in PS-NP exposed C. elegans. In PS-NP exposed C. elegans, expressions of 4 other germline hedgehog ligand genes and 10 hedgehog receptor genes were increased by daf-16 RNAi. Among these candidate genes, expressions of hedgehog ligand genes (grl-15, grl-16, qua-1, and wrt-1) and hedgehog receptor genes (ptr-23, scp-1, ptd-2, and ncr-1) could be increased by PS-NP (1–100 μg/L), and their transgenerational expressions were observed after PS-NP exposure. RNAi of grl-15, grl-16, qua-1, wrt-1, ptr-23, scp-1, ptd-2, and ncr-1 caused resistance to transgenerational PS-NP toxicity. In nematodes exposed to PS-NPs, RNAi of wrt-3, grl-15, grl-16, qua-1, and wrt-1 at parental generation (P0-G) inhibited expressions of ptr-23, scp-1, ptd-2, and ncr-1 in their offspring. Moreover, we observed increased expressions of insulin peptides genes (ins-3, ins-39, and daf-28) in PS-NP exposed daf-16(RNAi) nematodes, suggesting formation of feedback loop. We raise the molecular basis for formation of toxicity on multiple generations after nanoplastic exposure at P0-G.

Polystyrene nanoparticles regulate microbial stress response and cold adaptation in mainstream anammox process at low temperature

Nanoplastic pollution has become one of the most pressing environmental issues, and its bioaccumulation in aquatic environment also causes a great difficulty in treatment. Therefore, this work investigated the microbial dynamics of mainstream anaerobic ammonia oxidizing (anammox) process to treat the wastewater containing typical nanoplastics, as well as the fate and regulation mechanism of polystyrene nanoparticles (PS-NPs) with different concentrations. The results showed that 0.1–0.5 mg L-1 of PS-NPs had no significant effect on the nitrogen removal efficiency (NRE). When the concentration of PS-NPs increased from 0.5 mg L-1 to 2 mg L-1, the NRE of R1 with PS-NPs decreased from 94.9 ± 2.3 % to 77.0 ± 1.6 %, while the control reactor R0 maintained a stable NRE. Notably, the relative abundance of Ca. Kuenenia decreased from 17.4 % to 14.8 %, and that of Ca. Brocadia slightly decreased from 5.9 % to 5.0 % in R1. In addition, PS-NPs induced oxidative stress in anammox consortia, leading to the significant increase in reactive oxygen species (ROS) and lactate dehydrogenase (LDH) as well as cell membrane damage. PS-NPs also downregulated the content of heme c and further inhibited anammox activity. Based on the molecular docking simulation and western blotting, cold shock proteins (CSPs) could bind to PS-NPs and reduce the performance of anammox processes at low temperatures.

Mass Spectrometry-Based Top-Down Proteomics in Nanomedicine: Proteoform-Specific Measurement of Protein Corona.

Conventional mass spectrometry (MS)-based bottom-up proteomics (BUP) analysis of the protein corona [i.e., an evolving layer of biomolecules, mostly proteins, formed on the surface of nanoparticles (NPs) during their interactions with biomolecular fluids] enabled the nanomedicine community to partly identify the biological identity of NPs. Such an approach, however, fails to pinpoint the specific proteoforms─distinct molecular variants of proteins in the protein corona. The proteoform-level information could potentially advance the prediction of the biological fate and pharmacokinetics of nanomedicines. Recognizing this limitation, this study pioneers a robust and reproducible MS-based top-down proteomics (TDP) technique for characterizing proteoforms in the protein corona. Our TDP approach has successfully identified about 900 proteoforms in the protein corona of polystyrene NPs, ranging from 2 to 70 kDa, revealing proteoforms of 48 protein biomarkers with combinations of post-translational modifications, signal peptide cleavages, and/or truncations─details that BUP could not fully discern. This advancement in MS-based TDP offers a more advanced approach to characterize NP protein coronas, deepening our understanding of NPs' biological identities. We, therefore, propose using both TDP and BUP strategies to obtain more comprehensive information about the protein corona, which, in turn, can further enhance the diagnostic and therapeutic efficacy of nanomedicine technologies.

Protein corona alleviates adverse biological effects of nanoplastics in breast cancer cells.

Pollution from micro- and nanoplastics (MNPs) has long been a topic of concern due to its potential impact on human health. MNPs can circulate through human blood and, thus far, have been found in the lungs, spleen, stomach, liver, kidneys and even in the brain, placenta, and breast milk. While data are already available on the adverse biological effects of pristine MNPs (e.g. oxidative stress, inflammation, cytotoxicity, and even cancer induction), no report thus far clarified whether the same effects are modulated by the formation of a protein corona around MNPs. To this end, here we use pristine and human-plasma pre-coated polystyrene (PS) nanoparticles (NPs) and investigate them in cultured breast cancer cells both in terms of internalization and cell biochemical response to the exposure. It is found that pristine NPs tend to stick to the cell membrane and inhibit HER-2-driven signaling pathways, including phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways, which are associated with cancer cell survival and growth. By contrast, the formation of a protein corona around the same NPs can promote their uptake by endocytic vesicles and final sequestration within lysosomes. Of note is that such intracellular fate of PS-NPs is associated with mitigation of the biochemical alterations of the phosphorylated AKT (pAKT)/AKT and phosphorylated ERK (pERK)/ERK levels. These findings provide the distribution of NPs in human breast cancer cells, may broaden our understanding of the interactions between NPs and breast cancer cells and underscore the crucial role of the protein corona in modulating the impact of MNPs on human health.

Deep Plasma Proteome Profiling by Modulating Single Nanoparticle Protein Corona with Small Molecules.

The protein corona, a dynamic biomolecular layer that forms on nanoparticle (NP) surfaces upon exposure to biological fluids is emerging as a valuable diagnostic tool for improving plasma proteome coverage analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Here, we show that spiking small molecules, including metabolites, lipids, vitamins, and nutrients (namely, glucose, triglyceride, diglycerol, phosphatidylcholine, phosphatidylethanolamine, L-α-phosphatidylinositol, inosine 5'-monophosphate, and B complex), into plasma can induce diverse protein corona patterns on otherwise identical NPs, significantly enhancing the depth of plasma proteome profiling. The protein coronas on polystyrene NPs when exposed to plasma treated with an array of small molecules (n=10) allowed for detection of 1793 proteins marking an 8.25-fold increase in the number of quantified proteins compared to plasma alone (218 proteins) and a 2.63-fold increase relative to the untreated protein corona (681 proteins). Furthermore, we discovered that adding 1000 µg/ml phosphatidylcholine could singularly enable the detection of 897 proteins. At this specific concentration, phosphatidylcholine selectively depleted the four most abundant plasma proteins, including albumin, thus reducing the dynamic range of plasma proteome and enabling the detection of proteins with lower abundance. By employing an optimized data-independent acquisition (DIA) approach, the inclusion of phosphatidylcholine led to the detection of 1436 proteins in a single plasma sample. Our molecular dynamic results revealed that phosphatidylcholine interacts with albumin via hydrophobic interactions, h-bonds, and water-bridges. Addition of phosphatidylcholine also enabled the detection of 337 additional proteoforms compared to untreated protein corona using a top-down proteomics approach. These significant achievements are made utilizing only a single NP type and one small molecule to analyze a single plasma sample, setting a new standard in plasma proteome profiling. Given the critical role of plasma proteomics in biomarker discovery and disease monitoring, we anticipate widespread adoption of this methodology for identification and clinical translation of proteomic biomarkers into FDA approved diagnostics.

P01-36 Proeryptotic effects of non-functionalized polystyrene nanoparticles with different diameters

P01-36 Proeryptotic effects of non-functionalized polystyrene nanoparticles with different diameters

Optical trapping-induced crystallization promoted by gold and silicon nanoparticles.

This study investigates the promotion of sodium chlorate (NaClO3) crystallization through optical trapping, enhanced by the addition of gold nanoparticles (AuNPs) and silicon nanoparticles (SiNPs). Using a focused laser beam at the air-solution interface of a saturated NaClO3 solution with AuNPs or SiNPs, the aggregates of these particles were formed at the laser focus, the nucleation and growth of metastable NaClO3 (m-NaClO3) crystals were induced. Continued laser irradiation caused these m-NaClO3 crystals to undergo repeated cycles of growth and dissolution, eventually transitioning to a stable crystal form. Our comparative analysis showed that AuNPs, due to their significant heating due to higher photon absorption efficiency, caused more pronounced size fluctuations in m-NaClO3 crystals compared to the stable behavior observed with SiNPs. Interestingly, the maximum diameter of the m-NaClO3 crystals that appeared during the size fluctuation step was consistent, regardless of nanoparticle type, concentration, or size. The crystallization process was also promoted by using polystyrene nanoparticles, which have minimal heating and electric field enhancement, suggesting that the reduction in activation energy for nucleation at the particle surface is a key factor. These findings provide critical insights into the mechanisms of laser-induced crystallization, emphasizing the roles of plasmonic heating, particle surfaces, and optical forces.

Polystyrene nanoplastics enhance poxvirus preference for migrasome-mediated transmission

Since the emergence of a global outbreak of mpox in 2022, understanding the transmission pathways and mechanisms of Orthopoxviruses, including vaccinia virus (VACV), has become paramount. Nanoplastic pollution has become a significant global issue due to its widespread presence in the environment and potential adverse effects on human health. These emerging pollutants pose substantial risks to both living organisms and the environment, raising serious health concerns related to their proliferation. Despite this, the effects of nanoparticles on viral transmission dynamics remain unclear. This study explores how polystyrene nanoparticles (PS-NPs) influence the transmission of VACV through migrasomes. We demonstrate that PS-NPs accelerate the formation of migrasomes early in the infection process, facilitating VACV entry as soon as 15 h post-infection (hpi), compared to the usual onset at 36 hpi. Immunofluorescence and transmission electron microscopy (TEM) reveal significant co-localization of VACV with migrasomes induced by PS-NPs by 15 hpi. This interaction coincides with an increase in lipid droplet size, attributed to higher cholesterol levels influenced by PS-NPs. By 36 hpi, migrasomes exposed to both PS-NPs and VACV exhibit distinct features, such as retraction fibers and larger lipid droplets, emphasizing their critical role in cargo transport during viral infections. These results suggest that PS-NPs may act as modulators of viral transmission dynamics through migrasomes, with potential implications for antiviral strategies and environmental health.

Sub-ppm-level detection of nanoplastics using au nanograting and application to disposable plasticware

The detection of sub-ppm levels of nanoplastics using a surface-enhanced Raman scattering sensor was demonstrated. The proposed Au nanograting as surface-enhanced Raman scattering sensor, fabricated using nanoimprint lithography and oblique angle deposition, was designed to contain nanoplastics with enhanced local plasmonic fields. Electromagnetic simulations and Raman measurements show that the plasmonic field enhanced inside the nanograting enables the detection of Raman signal from 50-nm-sized polystyrene nanoparticles in 3-μL-droplets at a concertation of 0.1 ppm. Furthermore, the proposed sensor was applied to detect polystyrene in microwave-heated water in a disposable bowl. The results show that the proposed sensor offers promising potential for detecting nanoplastics released from plasticware.

Impact of diatomit on the transport behavior of unmodified and carboxyl-modified nanoplastics in saturated porous media

Due to the extensive use of plastic products and unreasonable disposal, nanoplastics contamination has become one of the important environmental problems that mankind must face. The composition and structure of porous media can determine the complexity and diversity of the transport behavior of nanoplastics. In this study, the influence of diatomite (DIA) on the nanoplastics transport in porous media is investigated by column experiments combined with XDLVO interaction energy and transport model. Results suggest that the recovery rates of unmodified polystyrene nanoparticles (PSNPs) and carboxyl-modified polystyrene nanoparticles (PSNPs-COOH) in the porous media containing DIA decreases compared with that in the pure quartz sand (QS), and the BTCs showed a “blocking” pattern. The presence of DIA inhibits the transport of both PSNPs and PSNPs-COOH, but the inhibition is not significant. This may be because the presence of DIA provides more favorable deposition sites for PSNPs and PSNPs-COOH to some extent. However, since DIA itself carries a certain negative charge, this can only play a role in compressing the double electric layer for PSNPs and PSNPs-COOH with the same negative charge, and cannot destabilize them. The migration capacity of PSNPs and PSNPs-COOH is strongest in the DIA-QS porous media at pH = 7, and is weak at pH = 9 and pH = 5. The inhibition of migration at pH = 9 can be attributed to the dissolution of the DIA surface under alkaline conditions and the formation of pore and defect structures, which provide more deposition sites for PSNPs and PSNPs-COOH. The presence of humic acid (HA) leads to an increase in the mobility of PSNPs and PSNPs-COOH, and the mobility is enhanced with HA concentration. The mobility of PSNPs and PSNPs-COOH in DIA-QS decreases with ionic valence and ionic strength, and PSNPs-COOH is more significantly inhibited compared to PSNPs.

Polystyrene nanoparticles with different particle sizes cause autophagy by ROS/ERS/FOXO1 axis in the Cyprinus carpio kidney affecting immunological function

Microplastic pollution poses challenges for ecosystems worldwide, and nanoplastics (NPs, 1–1000 nm) have been identified as persistent pollutants. However, although some studies have described the hazards of NPs to aquatic organisms, the toxicological processes of NPs in the common carp kidney and the biotoxicity of differently sized NPs remain unclear. In this study, we used juvenile common carp as an in vivo model that were constantly exposed to freshwater at 1000 μg/L polystyrene nanoparticle (PSNP) concentrations (50, 100, and 400 nm) for 28 days. Simultaneously, we constructed an in vitro model utilizing grass fish kidney cells (CIK) to study the toxicological effects of PSNPs of various sizes. We performed RT-PCR and Western blot assays on the genes involved in FOXO1, HMGB1, HIF-1α, endoplasmic reticulum stress, autophagy, and immunoreaction. According to these results, exposure to PSNPs increased reactive oxygen species (ROS) levels, and the carp kidneys experienced endoplasmic reticulum stress. Additionally, PSNPs promoted renal autophagy by activating the ROS/ERS/FOXO1 (ERS: endoplasmic reticulum stress) pathway, and it affected immunological function by stimulating the ROS/HMGB1/HIF-1α signaling pathway. This study provides new insights into the contamination hazards of NPs in freshwater environments, as well as the harm they pose to the human living environments. The relationship between particle size and the degree of damage caused by PSNPs to organisms is a potential future research direction.

Single-nanoparticle electrophoretic mobility determination and trapping using active-feedback 3D tracking.

Nanoparticles (NP) are versatile materials with widespread applications across medicine and engineering. Despite rapid incorporation into drug delivery, therapeutics, and many more areas of research and development, there is a lack of robust characterization methods. Light scattering techniques such as dynamic light scattering (DLS) and electrophoretic light scattering (ELS) use an ensemble-averaged approach to the characterization of nanoparticle size and electrophoretic mobility (EM), leading to inaccuracies when applied to polydisperse or heterogeneous populations. To address this lack of single-nanoparticle characterization, this work applies 3D Single-Molecule Active Real-time Tracking (3D-SMART) to simultaneously determine NP size and EM on a per-particle basis. Single-nanoparticle EM is determined by using active feedback to "lock on" to a single particle and apply an oscillating electric field along one axis. A maximum likelihood approach is applied to extract the single-particle EM from the oscillating nanoparticle position along the field-actuated axis, while mean squared displacement is used along the non-actuated axes to determine size. Unfunctionalized and carboxyl-functionalized polystyrene NPs are found to have unique EM based on their individual size and surface characteristics, and it is demonstrated that single-nanoparticle EM is a more precise tool for distinguishing unique NP preparations than diffusion alone, able to determine the charge number of individual NPs to an uncertainty of less than 30. This method also explored individual nanoparticle EM in various ionic strengths (0.25-5 mM) and found decreased EM as a function of increasing ionic strength, in agreement with results determined via bulk characterization methods. Finally, it is demonstrated that the electric field can be manipulated in real time in response to particle position, resulting in one-dimensional electrokinetic trapping. Critically, this new single-nanoparticle EM determination and trapping method does not require microfluidics, opening the possibility for the exploration of single-nanoparticle EM in live tissue and more comprehensive characterization of nanoparticles in biologically relevant environments.