Two-color interferometric scattering (iSCAT) microscopy reveals structural dynamics in discrete plasmonic molecules.

Plasmonic molecules are discrete assemblies of noble metal nanoparticles (NPs) that are of interest as transducers in optical nanosensors. So far, NPs with diameters of ∼40 nm have been the preferred building blocks for plasmonic molecules intended as optical single molecule sensors due to difficulties associated with detecting smaller NPs through elastic scattering in conventional darkfield microscopy. Here, we apply 405 nm, 445 nm two-color interferometric scattering (iSCAT) microscopy to characterize polyethylene glycol (PEG) tethered dimers of 10 nm and 20 nm Ag NPs and their monomers. Dimers of both NP sizes can be discerned from their respective monomers through changes in the average iSCAT contrast. In the case of 20 nm Ag NPs, dimer formation induces a change in the sign of the iSCAT contrast, providing a characteristic signal for detecting binding events. 20 nm Ag NP dimers with 0.4 kDa and 3.4 kDa polyethylene glycol (PEG) spacers show iSCAT contrast distributions with significantly different averages on both wavelength channels. The iSCAT contrast measured for individual PEG-tethered 10 nm or 20 nm NP dimers as a function of time shows contrast fluctuations indicative of a rich structural dynamics in the assembled plasmonic molecules, which provides an additional metric to discern dimers from monomers and paves the path to a new class of interferometric plasmon rulers.

Membrane fluidity properties of lipid-coated polylactic acid nanoparticles.

Lipid coating is considered a versatile strategy to equip nanoparticles (NPs) with a biomimetic surface coating, but the membrane properties of these nanoassemblies remain in many cases insufficiently understood. In this work, we apply C-Laurdan generalized polarization (GP) measurements to probe the temperature-dependent polarity of hybrid membranes consisting of a lipid monolayer adsorbed onto a polylactic acid (PLA) polymer core as function of lipid composition and compare the behavior of the lipid coated NPs (LNPs) with that of liposomes assembled from identical lipid mixtures. The LNPs were generated by nanoprecipitation of the polymer in aqueous solutions containing two types of lipid mixtures: (i) cholesterol, dipalmitoylphosphatidylcholine (DPPC), and the ganglioside GM3, as well as (ii) dioleoylphosphatidylcholine (DOPC), DPPC and GM3. LNPs were found to exhibit more distinct and narrower phase transitions than corresponding liposomes and to retain detectable phase transitions even for cholesterol or DOPC concentrations that yielded no detectable transitions in liposomes. These findings together with higher GP values in the case of the LNPs for temperatures above the phase transition temperature indicate a stabilization of the membrane through the polymer core. LNP binding studies to GM3-recognizing cells indicate that differences in the membrane fluidity affect binding avidity in the investigated model system.

Roadmap on optical sensors.

Optical sensors and sensing technologies are playing a more and more important role in our modern world. From micro-probes to large devices used in such diverse areas like medical diagnosis, defence, monitoring of industrial and environmental conditions, optics can be used in a variety of ways to achieve compact, low cost, stand-off sensing with extreme sensitivity and selectivity. Actually, the challenges to the design and functioning of an optical sensor for a particular application requires intimate knowledge of the optical, material, and environmental properties that can affect its performance. This roadmap on optical sensors addresses different technologies and application areas. It is constituted by twelve contributions authored by world-leading experts, providing insight into the current state-of-the-art and the challenges their respective fields face. Two articles address the area of optical fibre sensors, encompassing both conventional and specialty optical fibres. Several other articles are dedicated to laser-based sensors, micro- and nano-engineered sensors, whispering-gallery mode and plasmonic sensors. The use of optical sensors in chemical, biological and biomedical areas is discussed in some other papers. Different approaches required to satisfy applications at visible, infrared and THz spectral regions are also discussed.

Two-Color iSCAT Imaging of Ag Nanoparticles Resolves Size and Ambient Refractive Index Changes.

The ability to discern noble metal nanoparticles (NPs) with different sizes and in ambient media with different refractive indices has important applications in imaging and sensing. Here a two-color (405 nm, 445 nm) interferometric scattering (iSCAT) detection scheme is applied to characterize the wavelength-dependent iSCAT contrast of Ag NPs with nominal diameters of 10, 20, 40, and 60 nm and to distinguish between NPs of different sizes. The iSCAT contrast also depends on the ambient refractive index and the relative iSCAT contrast on both channels revealed a spectral red-shift for 40 and 60 nm Ag NPs when the ambient refractive index was increased from n = 1.3892 to n = 1.4328. With the selected wavelength channels, the spectral resolution of the two-color imaging strategy was, however, insufficient to resolve spectral shifts induced by refractive index changes for 10 and 20 nm Ag NPs.

Nanoplastics Weathering and Polycyclic Aromatic Hydrocarbon Mobilization.

Despite increasing efforts to recycle plastic materials, large quantities of plastics waste continue to accumulate in the oceans. Persistent mechanical and photochemical degradation of plastics in the oceans yields micro- and nanoscale plastic particles, which represent potential vectors for mobilizing hydrophobic carcinogens in an aqueous milieu. Yet, the fate and potential threats associated with plastics remain largely unexplored. Herein, we apply an accelerated weathering protocol to consumer plastics to characterize the effect of photochemical weathering on the size, morphology, and chemical composition of nanoplastics under defined conditions and validate that the photochemical degradation is consistent with plastics harvested from the Pacific Ocean. Machine learning algorithms trained with accelerated weathering data successfully classify weathered plastics from nature. We demonstrate that photodegradation of poly(ethylene terephthalate) (PET)-containing plastics produces enough CO2 to induce a mineralization process that results in the deposition of CaCO3 on nanoplastics. Finally, we determine that despite UV-radiation induced photochemical degradation and mineral deposition, nanoplastics retain their ability to sorb, mobilize, and increase bioaccessibility of polycyclic aromatic hydrocarbons (PAHs) in water and under simulated physiological gastric and intestinal conditions.

Ganglioside-Functionalized Nanoparticles for Chimeric Antigen Receptor T-Cell Activation at the Immunological Synapse.

Chimeric Antigen Receptor (CAR) T cell therapy has proven to be an effective strategy against hematological malignancies but persistence and activity against solid tumors must be further improved. One emerging strategy for enhancing efficacy is based on directing CAR T cells to antigen presenting cells (APCs). Activation of CAR T cells at the immunological synapse (IS) formed between APC and T cell is thought to promote strong, persistent antigen-specific T cell-mediated immune responses but requires integration of CAR ligands into the APC/T-cell interface. Here, we demonstrate that CAR ligand functionalized, lipid-coated, biodegradable polymer nanoparticles (NPs) that contain the ganglioside GM3 (GM3-NPs) bind to CD169 (Siglec-1)-expressing APCs and localize to the cell contact site between APCs and CAR T cells upon initiation of cell conjugates. The CD169+ APC/CAR T-cell interface is characterized by a strong optical colocalization of GM3-NPs and CARs, enrichment of F-actin, and recruitment of ZAP-70, indicative of integration of GM3-NPs into a functional IS. Ligands associated with GM3-NPs localized to the APC/T-cell contact site remain accessible to CARs and result in robust T-cell activation. Overall, this work identifies GM3-NPs as a potential antigen delivery platform for active targeting of CD169 expressing APCs and enhancement of CAR T-cell activation at the NP-containing IS.

Plasmonic Enhancement Strategies for Light-Driven Microbe Inactivation.

Light can be an effective antimicrobial. UV-C light, in particular, is now commonly used to sterilize inanimate surfaces, water, and even air. Highly energetic light can, however, also lead to unwanted photodamage and be hazardous. Consequently, conventional light-mediated microbe inactivation is not suitable for all applications. Plasmonic nanostructures can enhance electromagnetic fields in the visible range of the electromagnetic spectrum and show unique light-induced responses that can drive strong antimicrobial effects even for wavelengths that without plasmonic enhancement have little to no antimicrobial impact. Plasmonic nanostructures offer thus a potential strategy to expand the antimicrobial effect of light to wavelength and intensity ranges in which light-associated collateral damages are lower. This Perspective examines selected plasmon-enhanced antimicrobial strategies, elucidates the underlying physico-chemical mechanisms, and discusses applications.

Characterizing Lipid-Coated Mesoporous Silica Nanoparticles as CD169-Binding Delivery System for Rilpivirine and Cabotegravir.

Herein, lipid-coated mesoporous silica nanoparticles (LMSN) are investigated as biomimetic delivery vehicle for two antiretroviral compounds (ARVs), rilpivirine (RPV) and cabotegravir (CAB). Monosialodihexosylganglioside (GM3) is incorporated into the membrane to facilitate LMSN binding to CD169 (Siglec-1)-expressing myeloid cells, that are predominantly expressed in secondary lymphoid tissues in vivo. It is demonstrated that in addition to providing CD169-binding functionalities, the lipid membrane around the silica core provides stealth properties that dampen the inflammatory cytokine response to ARVs-loaded LMSN in human monocyte-derived macrophages. Quantification of RPV and CAB releases from nanoparticles, and assessment of antiviral potency to human immunodeficiency virus (HIV-1) infection in vitro reveals that RPV and CAB co-formulated into LMSN retain optimal antiviral potency for 90 days, even upon storage at room temperature, making LMSN an attractive nanoplatform, immune to cold chain requirements. These findings suggest that GM3-LMSN equip the mesoporous silica nanoparticle (MSN) core with lipid-derived properties for surface passivation and lipid-mediated binding that are of high interest for achieving an effective delivery of ARVs to tissue reservoirs of HIV-1 replication.

Multivalent Ligand-Nanoparticle Conjugates Amplify Reactive Oxygen Species Second Messenger Generation and Enhance Epidermal Growth Factor Receptor Phosphorylation.

The epidermal growth factor (EGF) receptor (EGFR) is heterogeneously distributed on the cellular surface and enriched in clusters with diameters of tens of nanometers. Multivalent presentation of EGF ligand on nanoparticles (NPs) provides an approach for controlling and amplifying the local activation of EGFR in these clusters. Reactive oxygen species (ROS) have been indicated to play a role in the regulation of EGFR activation as second messengers, but the effect of nanoconjugation on EGF-mediated ROS formation and ROS-induced EGFR activation is not well established. The goal of this manuscript is to characterize the multivalent enhancement of EGF-induced ROS formation and to test its effect on EGFR phosphorylation in breast cancer cell models using gold (Au) NPs with a diameter of 81 ± 1 nm functionalized with two different EGF ligand densities (12 ± 7 EGF/NP (NP-EGF12) and 87 ± 6 EGF/NP (NP-EGF87)). In the EGFR overexpressing cell lines MDA-MB-231 and MDA-MB-468, NP-EGF87 achieved a measurable multivalent enhancement of ROS that peaked at concentrations c ROSmax ≤ 25 pM and that were EGFR and nicotinamide adenine dinucleotide phosphate oxidase (NOX) dependent. NP-EGF12 failed to generate comparable ROS levels as NP-EGF87 in the investigated NP input concentration range (0-100 pM). In cells with nearly identical numbers of bound NP-EGF87 and NP-EGF12, the ROS levels for NP-EGF87 were systematically higher, indicating that the multivalent enhancement is exclusively related not only to avidity but also to a stronger stimulation per NP. Importantly, the increase in EGF-induced ROS formation associated with EGF nanoconjugation at c ROSmax resulted in a measurable gain in EGFR phosphorylation, confirming that ROS generation contributes to the multivalent enhancement of EGFR activation in response to NP-EGF87.

Plasmonic photoreactors-coated plastic tubing as combined-active-and-passive antimicrobial flow sterilizer.

Plastic materials are ubiquitous in medical devices and consumer goods. As bacterial contamination of plastic surfaces can pose significant health risks, there is a need for effective approaches both to inactivate bacteria on plastic surfaces and to prevent colonization of plastic surfaces. In this study, we evaluate a plasmonic photoreactor coating for plastic surfaces that provides both active and passive antimicrobial effects and implement a visible light-driven antibacterial flow sterilizer. We demonstrate that this approach inactivates bacteria in an aqueous suspension passed through a photoreactor-coated polyethylene tubing, achieving log reduction values (LRVs) > 5 for both Gram-positive and -negative bacteria under resonant LED illumination. Importantly, the antimicrobial flow sterilizers do not cause a detectable loss of functionality for monoclonal antibodies that were included in this work as an example of high-value biologics that require sterilization. Under ambient light illumination, the plasmonic photoreactor coating exhibits a significant inhibitory effect on bacterial colonization and biofilm formation. The inhibitory effect was substantially weaker for mammalian cells, indicating some selectivity in the protection provided by the coating.

Wavelength-Dependent Bifunctional Plasmonic Photocatalysis in Au/Chalcopyrite Hybrid Nanostructures.

Excited, or "hot" charge carrier generation and transfer driven by the decay of localized surface plasmon resonances (LSPRs) are key steps in plasmonic photocatalysis. Hybrid structures that contain both metal and semiconductor building blocks facilitate the extraction of reactive charge carriers and their utilization for photoelectrocatalysis. Additional functionality arises from hybrid structures that combine noble metal nanostructures with semiconductor components, such as chalcopyrite (CuFeS2) nanocrystals (NCs), which by themselves support quasistatic resonances. In this work, we use a hybrid membrane to integrate Au nanorods (NRs) with a longitudinal LSPR at 745 nm and CuFeS2 NCs with a resonance peak at 490 nm into water-stable nanocomposites for robust and bifunctional photocatalysis of oxygen and hydrogen evolution reactions in a wavelength-dependent manner. Excitation of NRs or NCs in the nanocomposite correlates with increased hydrogen or oxygen evolution, respectively, consistent with a light-driven electron transfer between the metal and semiconductor building blocks, the direction of which depends on the wavelength. The bifunctional photoreactivity of the nanocomposite is enhanced by Cu(I)/Cu(II)-assisted catalysis on the surface of the NCs.

Virus-Mimicking Polymer Nanoparticles Targeting CD169+ Macrophages as Long-Acting Nanocarriers for Combination Antiretrovirals.

Monosialodihexosylganglioside (GM3)-presenting lipid-coated polymer nanoparticles (NPs) that recapitulate the sequestration of human immunodeficiency virus-1 (HIV-1) particles in CD169+ virus-containing compartments (VCCs) of macrophages were developed as carriers for delivery and sustained release of a combination of two antiretrovirals (ARVs), rilpivirine (RPV) and cabotegravir (CAB). RPV and CAB were co-loaded into GM3-presenting lipid-coated polylactic acid (PLA) and poly(lactic-co-glycolic acid) (PLGA) NPs without loss in potency of the drugs. GM3-presenting PLA NPs demonstrated the most favorable release properties and achieved inhibition of HIV-1 infection of primary human macrophages for up to 35 days. Intracellular localization of GM3-presenting PLA NPs in VCCs correlated with retention of intracellular ARV concentrations and sustained inhibition of HIV-1 infection. This work elucidates the design criteria of lipid-coated polymer NPs to utilize CD169+ macrophages as cellular drug depots for eradicating the viral reservoir sites or to achieve long-acting prophylaxis against HIV-1 infection.

Interfacial hydration determines orientational and functional dimorphism of sterol-derived Raman tags in lipid-coated nanoparticles.

Lipid-coated noble metal nanoparticles (L-NPs) combine the biomimetic surface properties of a self-assembled lipid membrane with the plasmonic properties of a nanoparticle (NP) core. In this work, we investigate derivatives of cholesterol, which can be found in high concentrations in biological membranes, and other terpenoids, as tunable, synthetic platforms to functionalize L-NPs. Side chains of different length and polarity, with a terminal alkyne group as Raman label, are introduced into cholesterol and betulin frameworks. The synthesized tags are shown to coexist in two conformations in the lipid layer of the L-NPs, identified as "head-out" and "head-in" orientations, whose relative ratio is determined by their interactions with the lipid-water hydrogen-bonding network. The orientational dimorphism of the tags introduces orthogonal functionalities into the NP surface for selective targeting and plasmon-enhanced Raman sensing, which is utilized for the identification and Raman imaging of epidermal growth factor receptor-overexpressing cancer cells.

Plasmonic nano-antimicrobials: properties, mechanisms and applications in microbe inactivation and sensing.

Bacterial, viral and fungal infections pose serious threats to human health and well-being. The continuous emergence of acute infectious diseases caused by pathogenic microbes and the rapid development of resistances against conventional antimicrobial drugs necessitates the development of new and effective strategies for the safe elimination of microbes in water, food or on surfaces, as well as for the inactivation of pathogenic microbes in human hosts. The need for new antimicrobials has triggered the development of plasmonic nano-antimicrobials that facilitate both light-dependent and -independent microbe inactivation mechanisms. This review introduces the relevant photophysical mechanisms underlying these plasmonic nano-antimicrobials, and provides an overview of how the photoresponses and materials properties of plasmonic nanostructures can be applied in microbial pathogen inactivation and sensing applications. Through a systematic analysis of the inactivation efficacies of different plasmonic nanostructures, this review outlines the current state-of-the-art in plasmonic nano-antimicrobials and defines the application space for different microbial inactivation strategies. The advantageous optical properties of plasmonic nano-antimicrobials also enhance microbial detection and sensing modalities and thus help to avoid exposure to microbial pathogens. Sensitive and fast plasmonic microbial sensing modalities and their theranostic and targeted therapeutic applications are discussed.

Stiffness of HIV-1 Mimicking Polymer Nanoparticles Modulates Ganglioside-Mediated Cellular Uptake and Trafficking.

The monosialodihexosylganglioside, GM3, and its binding to CD169 (Siglec-1) have been indicated as key factors in the glycoprotein-independent sequestration of the human immunodeficiency virus-1 (HIV-1) in virus-containing compartments (VCCs) in myeloid cells. Here, lipid-wrapped polymer nanoparticles (NPs) are applied as a virus-mimicking model to characterize the effect of core stiffness on NP uptake and intracellular fate triggered by GM3-CD169 binding in macrophages. GM3-functionalized lipid-wrapped NPs are assembled with poly(lactic-co-glycolic) acid (PLGA) as well as with low and high molecular weight polylactic acid (PLAlMW and PLAhMW) cores. The NPs have an average diameter of 146 ± 17 nm and comparable surface properties defined by the self-assembled lipid layer. Due to differences in the glass transition temperature, the Young's modulus (E) differs substantially under physiological conditions between PLGA (E PLGA = 60 ± 32 MPa), PLAlMW (E PLA lMW = 86 ± 25 MPa), and PLAhMW (E PLA hMW = 1.41 ± 0.67 GPa) NPs. Only the stiff GM3-presenting PLAhMW NPs but not the softer PLGA or PLAlMW NPs avoid a lysosomal pathway and localize in tetraspanin (CD9)-positive compartments that resemble VCCs. These observations suggest that GM3-CD169-induced sequestration of NPs in nonlysosomal compartments is not entirely determined by ligand-receptor interactions but also depends on core stiffness.

Physiologically Relevant Mechanics of Biodegradable Polyester Nanoparticles.

Despite the extensive use of biodegradable polyester nanoparticles for drug delivery, and reports of the strong influence of nanoparticle mechanics on nano-bio interactions, there is a lack of systematic studies on the mechanics of these nanoparticles under physiologically relevant conditions. Here, we report indentation experiments on poly(lactic acid) and poly(lactide-co-glycolide) nanoparticles using atomic force microscopy. While dried nanoparticles were found to be rigid at room temperature, their elastic modulus was found to decrease by as much as 30 fold under simulated physiological conditions (i.e., in water at 37 °C). Differential scanning calorimetry confirms that this softening can be attributed to the glass transition of the nanoparticles. Using a combination of mechanical and thermoanalytical characterization, the plasticizing effects of miniaturization, molecular weight, and immersion in water were investigated. Collectively, these experiments provide insight for experimentalists exploring the relationship between polymer nanoparticle mechanics and in vivo behavior.

Evolution of near- and far-field optical properties of Au bipyramids upon epitaxial deposition of Ag.

Bimetallic plasmonic nanostructures provide composition and spatial distribution of the individual components in the nanostructure in addition to overall size and morphology as degrees of freedom for tuning near- and far-field optical responses. AgAuAg nanorods (NRs) generated through epitaxial deposition of Ag on the tips of Au bipyramids (BPs) are an important bimetallic model system whose longitudinal dipolar plasmon mode first shows a spectral blue-shift upon initial deposition of Ag on the Au BP tips followed by a red-shift after additional deposition of Ag. Here, we quantify the relative contributions from morphological and compositional effects to the far-field spectral shift of the longitudinal and vertical dipolar plasmon modes during the initial deposition of Ag and compare the near-field in Ag and AgAuAg NRs with lengths between L = 130 nm-280 nm under whitelight illumination through electromagnetic simulations. Subsequently, we experimentally characterize the near-field around AgAuAg NRs with lengths between L = 88.1-749.0 nm at a constant excitation wavelength of 1064 nm on a silicon (Si) support through scattering type near-field scanning microscopy (sNSOM). We detect Fabry-Perot resonance-like higher order multipolar plasmon resonances whose order and near-field pattern depends on the length and composition of the NRs as well as the refractive index of the ambient medium. We find that under oblique illumination higher order multipolar modes with an even symmetry dominate on the high refractive index Si substrate due to strong electromagnetic interactions between the NR and the substrate.

Hybrid Plasmonic Photoreactors as Visible Light-Mediated Bactericides.

Photocatalytic compounds and complexes, such as tris(bipyridine)ruthenium(II), [Ru(bpy)3]2+, have recently attracted attention as light-mediated bactericides that can help to address the need for new antibacterial strategies. We demonstrate in this work that the bactericidal efficacy of [Ru(bpy)3]2+ and the control of its antibacterial function can be significantly enhanced through combination with a plasmonic nanoantenna. We report strong, visible light-controlled bacterial inactivation with a nanocomposite design that incorporates [Ru(bpy)3]2+ as a photocatalyst and a Ag nanoparticle (NP) core as a light-concentrating nanoantenna into a plasmonic hybrid photoreactor. The hybrid photoreactor platform is facilitated by a self-assembled lipid membrane that encapsulates the Ag NP and binds the photocatalyst. The lipid membrane renders the nanocomposite biocompatible in the absence of resonant illumination. Upon illumination, the plasmon-enhanced photoexcitation of the metal-to-ligand charge-transfer band of [Ru(bpy)3]2+ prepares the reactive excited state of the complex that oxidizes the nanocomposite membrane and increases its permeability. The photooxidation induces the release of [Ru(bpy)3]2+, Ag+, and peroxidized lipids into the ambient medium, where they interact synergistically to inactivate bacteria. We measured a 7 order of magnitude decrease in Gram-positive Arthrobacter sp. and a 4 order of magnitude decrease in Gram-negative Escherichia coli colony forming units with the photoreactor bactericides after visible light illumination for 1 h. In both cases, the photoreactor exceeds the bactericidal standard of a log reduction value of 3 and surpasses the antibacterial effect of free Ag NPs or [Ru(bpy)3]2+ by >4 orders of magnitude. We also implement the inactivation of a bacterial thin film in a proof-of-concept study.

Characterizing Large-Scale Receptor Clustering on the Single Cell Level: A Comparative Plasmon Coupling and Fluorescence Superresolution Microscopy Study.

Spatial clustering of cell membrane receptors has been indicated to play a regulatory role in signal initiation, and the distribution of receptors on the cell surface may represent a potential biomarker. To realize its potential for diagnostic purposes, scalable assays capable of mapping spatial receptor heterogeneity with high throughput are needed. In this work, we use gold nanoparticle (NP) labels with an average diameter of 72.17 ± 2.16 nm as bright markers for large-scale epidermal growth factor receptor (EGFR) clustering in hyperspectral plasmon coupling microscopy and compare the obtained clustering maps with those obtained through fluorescence superresolution microscopy (direct stochastic optical reconstruction microscopy, dSTORM). Our dSTORM experiments reveal average EGFR cluster sizes of 172 ± 99 and 150 ± 90 nm for MDA-MB-468 and HeLa, respectively. The cluster sizes decrease after EGFR activation. Hyperspectral imaging of the NP labels shows that differences in the EGFR cluster sizes are accompanied by differences in the average separations between electromagnetically coupled NPs. Because of the distance dependence of plasmon coupling, changes in the average interparticle separation result in significant spectral shifts. For the experimental conditions investigated in this work, hyperspectral plasmon coupling microscopy of NP labels identified the same trends in large-scale EGFR clustering as dSTORM, but the NP imaging approach provided the information in a fraction of the time. Both dSTORM and hyperspectral plasmon coupling microscopy confirm the cortical actin network as one structural component that determines the average size of EGFR clusters.