Plasmonic dielectric antennas for hybrid optical nanotweezing and optothermoelectric manipulation of single nanosized extracellular vesicles.

This paper showcases an experimental demonstration of near-field optical trapping and dynamic manipulation of an individual extracellular vesicle. This is accomplished through the utilization of a plasmonic dielectric nanoantenna designed to support an optical anapole state-a non-radiating optical state resulting from the destructive interference between electric and toroidal dipoles in the far-field, leading to robust near-field enhancement. To further enhance the field intensity associated with the optical anapole state, a plasmonic mirror is incorporated, thereby boosting trapping capabilities. In addition to demonstrating near-field optical trapping, the study achieves dynamic manipulation of extracellular vesicles by harnessing the thermoelectric effect. This effect is induced in the presence of an ionic surfactant, cetyltrimethylammonium chloride (CTAC), combined with plasmonic heating. Furthermore, the thermoelectric effect improves trapping stability by introducing a wide and deep trapping potential. In summary, our hybrid plasmonic-dielectric trapping platform offers a versatile approach for actively transporting, stably trapping, and dynamically manipulating individual extracellular vesicles.

Evaluating the glymphatic system via magnetic resonance diffusion tensor imaging along the perivascular spaces in brain tumor patients.

PURPOSE: To investigate glymphatic system function in patients with brain tumors, including both primary and secondary tumors, using diffusion tensor imaging along perivascular spaces (DTI-ALPS). METHODS: We retrospectively analyzed the MR DTI of 24 patients with unilateral brain tumors and compared them with age and sex-matched controls. We compared the DTI-ALPS index of the ipsi- and contralateral brain hemispheres. The region of interest was placed in the periventricular vessels adjacent to the lateral ventricles. Differences between sex, age, and kind of tumor (primary or brain metastasis) were evaluated. Correlations between DTI-ALPS index and age and the tumor's apparent diffusion coefficient (ADC) were also investigated. RESULTS: The DTI-ALPS index was significantly lower (p < 0.05) in the tumor-affected hemisphere (mean = 1.26 ± 0.24) than contralateral (mean = 1.43 ± 0.28). A comparison with healthy controls revealed no significant difference on the matched ipsilateral side. However, the DTI-ALPS index of the contralateral side of the patients was larger than the HC. Additionally, no statistically significant differences were found when analyzing the DTI-ALPS index vs. age, sex, and tumor entity. Additionally, we did not find a correlation between the DTI-ALPS index and patient age or tumor ADC. CONCLUSION: The decreased DTI-ALPS index in the tumor-affected hemisphere may be related to impaired glymphatic system function. However, cancer is often a systemic disease; thus, the DTI-ALPS index from the contralateral brain hemisphere may not generally be considered as a normal control. Nonetheless, the DTI-ALPS index does not only reflect diffusion in the perivascular spaces but it can also be influenced by factors such as axonal degeneration. Therefore, it does not directly reflect brain waste clearance and changes in the index should be interpreted carefully.

Towards rapid colorimetric detection of extracellular vesicles using optofluidics-enhanced color-changing optical metasurface.

Efficient transportation and delivery of analytes to the surface of optical sensors are crucial for overcoming limitations in diffusion-limited transport and analyte sensing. In this study, we propose a novel approach that combines metasurface optics with optofluidics-enabled active transport of extracellular vesicles (EVs). By leveraging this combination, we show that we can rapidly capture EVs and detect their adsorption through a color change generated by a specially designed optical metasurface that produces structural colors. Our results demonstrate that the integration of optofluidics and metasurface optics enables spectrometer-less and label-free colorimetric read-out for EV concentrations as low as 107 EVs/ml, achieved within a short incubation time of two minutes.

How do Antimicrobial Peptides Interact with the Outer Membrane of Gram-Negative Bacteria? Role of Lipopolysaccharides in Peptide Binding, Anchoring, and Penetration.

Gram-negative bacteria possess a complex structural cell envelope that constitutes a barrier for antimicrobial peptides that neutralize the microbes by disrupting their cell membranes. Computational and experimental approaches were used to study a model outer membrane interaction with an antimicrobial peptide, melittin. The investigated membrane included di[3-deoxy-d-manno-octulosonyl]-lipid A (KLA) in the outer leaflet and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) in the inner leaflet. Molecular dynamics simulations revealed that the positively charged helical C-terminus of melittin anchors rapidly into the hydrophilic headgroup region of KLA, while the flexible N-terminus makes contacts with the phosphate groups of KLA, supporting melittin penetration into the boundary between the hydrophilic and hydrophobic regions of the lipids. Electrochemical techniques confirmed the binding of melittin to the model membrane. To probe the peptide conformation and orientation during interaction with the membrane, polarization modulation infrared reflection absorption spectroscopy was used. The measurements revealed conformational changes in the peptide, accompanied by reorientation and translocation of the peptide at the membrane surface. The study suggests that melittin insertion into the outer membrane affects its permeability and capacitance but does not disturb the membrane's bilayer structure, indicating a distinct mechanism of the peptide action on the outer membrane of Gram-negative bacteria.

Mirror-Enhanced Plasmonic Nanoaperture for Ultrahigh Optical Force Generation with Minimal Heat Generation.

Double Nanohole Plasmonic Tweezers (DNH) have emerged as a powerful approach for confining light to sub-wavelength volume, enabling the trapping of nanoscale particles much smaller than the wavelength of light. However, to circumvent plasmonic heating effects, DNH tweezers are typically operated off-resonance, resulting in reduced optical forces and field enhancements. In this study, we introduce a novel DNH design with a reflector layer, enabling on-resonance illumination while minimizing plasmonic heating. This design efficiently dissipates heat and redistributes the electromagnetic hotspots, making them more accessible for trapping nanoscale particles and enhancing light-matter interactions. We also demonstrate low-power trapping and release of small extracellular vesicles. Our work opens new possibilities for trapping-assisted Surface Enhanced Raman Spectroscopy (SERS), plasmon-enhanced imaging, and single photon emission applications that demand strong light-matter interactions.

Scalable trapping of single nanosized extracellular vesicles using plasmonics.

Heterogeneous nanoscale extracellular vesicles (EVs) are of significant interest for disease detection, monitoring, and therapeutics. However, trapping these nano-sized EVs using optical tweezers has been challenging due to their small size. Plasmon-enhanced optical trapping offers a solution. Nevertheless, existing plasmonic tweezers have limited throughput and can take tens of minutes for trapping for low particle concentrations. Here, we present an innovative approach called geometry-induced electrohydrodynamic tweezers (GET) that overcomes these limitations. GET generates multiple electrohydrodynamic potentials, allowing parallel transport and trapping of single EVs within seconds. By integrating nanoscale plasmonic cavities at the center of each GET trap, single EVs can be placed near plasmonic cavities, enabling instant plasmon-enhanced optical trapping upon laser illumination without detrimental heating effects. These non-invasive scalable hybrid nanotweezers open new horizons for high-throughput tether-free plasmon-enhanced single EV trapping and spectroscopy. Other potential areas of impact include nanoplastics characterization, and scalable hybrid integration for quantum photonics.

Anapole-Assisted Low-Power Optical Trapping of Nanoscale Extracellular Vesicles and Particles.

This study addresses the challenge of trapping nanoscale biological particles using optical tweezers without the photothermal heating effect and the limitation presented by the diffraction limit. Optical tweezers are effective for trapping microscopic biological objects but not for nanoscale specimens due to the diffraction limit. To overcome this, we present an approach that uses optical anapole states in all-dielectric nanoantenna systems on distributed Bragg reflector substrates to generate strong optical gradient force and potential on nanoscale biological objects with negligible temperature rise below 1 K. The anapole antenna condenses the accessible electromagnetic energy to scales as small as 30 nm. Using this approach, we successfully trapped nanosized extracellular vesicles and supermeres (approximately 25 nm in size) using low laser power of only 10.8 mW. This nanoscale optical trapping platform has great potential for single molecule analysis while precluding photothermal degradation.

Optofluidic transport and assembly of nanoparticles using an all-dielectric quasi-BIC metasurface.

Manipulating fluids by light at the micro/nanoscale has been a long-sought-after goal for lab-on-a-chip applications. Plasmonic heating has been demonstrated to control microfluidic dynamics due to the enhanced and confined light absorption from the intrinsic losses of metals. Dielectrics, the counterpart of metals, has been used to avoid undesired thermal effects due to its negligible light absorption. Here, we report an innovative optofluidic system that leverages a quasi-BIC-driven all-dielectric metasurface to achieve subwavelength scale control of temperature and fluid motion. Our experiments show that suspended particles down to 200 nanometers can be rapidly aggregated to the center of the illuminated metasurface with a velocity of tens of micrometers per second, and up to millimeter-scale particle transport is demonstrated. The strong electromagnetic field enhancement of the quasi-BIC resonance increases the flow velocity up to three times compared with the off-resonant situation by tuning the wavelength within several nanometers range. We also experimentally investigate the dynamics of particle aggregation with respect to laser wavelength and power. A physical model is presented and simulated to elucidate the phenomena and surfactants are added to the nanoparticle colloid to validate the model. Our study demonstrates the application of the recently emerged all-dielectric thermonanophotonics in dealing with functional liquids and opens new frontiers in harnessing non-plasmonic nanophotonics to manipulate microfluidic dynamics. Moreover, the synergistic effects of optofluidics and high-Q all-dielectric nanostructures hold enormous potential in high-sensitivity biosensing applications.

Exosomes trapping, manipulation and size-based separation using opto-thermo-electrohydrodynamic tweezers.

Owing to the heterogeneity of exosomes in size and biomolecular composition, there is a need for new approaches for trapping, manipulating, and sorting of single exosomes in solution. Due to their small size ranging from 30 nm to 150 nm and their relatively low refractive index, their stable trapping using optical tweezers has been met with challenges. Trapping exosomes in an optical trap requires nearly 100 mW of input power, which predisposes them to photo-induced damage and membrane rupture at the laser focus. Here, we report a high stability opto-thermo-electrohydrodynamic tweezer for the stable stand-off trapping of single exosomes based on a concentric nanohole array (CNA) using laser illumination and an a.c. field. The CNA system generates two regions of electrohydrodynamic potentials several microns away from the laser focus where single exosomes are trapped. We demonstrate the rapid trapping within seconds, and selective dynamic manipulation of exosomes based on size using only 4.2 mW of input laser power. The proposed platform opens up a promising approach for stabilizing single exosomes in solution and controlling their distribution based on size without the risk of photo-induced damage.

High-speed nanoscale optical trapping with plasmonic double nanohole aperture.

Optical trapping with plasmonic double nanohole (DNH) apertures has proven to be an efficient method for trapping sub-50 nm particles due to their suppressed plasmonic heating effect and very high electric field enhancement in the gap region of the aperture. However, plasmonic tweezers are generally diffusion-limited, requiring particles to diffuse down to a few tens of nanometres from the high field enhancement regions before they can be trapped. The loading of target particles to the plasmonic hotspots can take several minutes for diluted samples. In this work, rapid particle transport and trapping of a 25 nm polystyrene sphere is demonstrated, leveraging an electrothermoplasmonic flow induced upon application of an AC field in the presence of a laser-induced temperature gradient. Using this approach, we demonstrate the rapid transport of a 25 nm polystyrene particle across a distance of 63 µm and trapping at the DNH under 16 s. This platform shows great potential for applications involving simultaneous trapping and plasmon-enhanced spectroscopies, such as Raman enhancement via the intense electric field enhancement in the DNH gap.

Multiplexed Long-Range Electrohydrodynamic Transport and Nano-Optical Trapping with Cascaded Bowtie Photonic Crystal Nanobeams.

Photonic crystal cavities with bowtie defects that combine ultrahigh Q and ultralow mode volume are theoretically studied for low-power nanoscale optical trapping. By harnessing the localized heating of the water layer near the bowtie region, combined with an applied alternating current electric field, this system provides long-range electrohydrodynamic transport of particles with average radial velocities of 30 µm/s towards the bowtie region on demand by switching the input wavelength. Once transported to a given bowtie region, synergistic interaction of optical gradient and attractive negative thermophoretic forces stably trap a 10 nm quantum dot in a potential well with a depth of 10 k_{B}T using a mW input power.

On-chip integrated quantum emitter with 'trap-enhance-guide': a simulation approach.

To address the challenges of developing a scalable system of an on-chip integrated quantum emitter, we propose to leverage the loss in our hybrid plasmonic-photonic structure to simultaneously achieve Purcell enhancement as well as on-chip maneuvering of nanoscale emitter via optical trapping with guided excitation-emission routes. In this report, we have analyzed the feasibility of the functional goals of our proposed system in the metric of trapping strength (∼8KBT), Purcell factor (>1000∼), and collection efficiency (∼10%). Once realized, the scopes of the proposed device can be advanced to develop a scalable platform for integrated quantum technology.

Engineering Electromagnetic Field Distribution and Resonance Quality Factor Using Slotted Quasi-BIC Metasurfaces.

Dielectric metasurfaces governed by bound states in the continuum (BIC) are actively investigated for achieving high-quality factors and strong electromagnetic field enhancements. Traditional approaches reported for tuning the performance of quasi-BIC metasurfaces include tuning the resonator size, period, and structure symmetry. Here we propose and experimentally demonstrate an alternative approach through engineering slots within a zigzag array of elliptical silicon resonators. Through analytical theory, three-dimensional electromagnetic modeling, and infrared spectroscopy, we systematically investigate the spectral responses and field distributions of the slotted metasurface in the mid-IR. Our results show that by introducing slots, the electric field intensity enhancement near the apex and the quality factor of the quasi-BIC resonance are increased by a factor of 2.1 and 3.3, respectively, in comparison to the metasurface without slots. Furthermore, the slotted metasurface also provides extra regions of electromagnetic enhancement and confinement, which holds enormous potential in particle trapping, sensing, and emission enhancement.

Transferring Microclusters of P. aeruginosa Biofilms to the Air-Liquid Interface of Bronchial Epithelial Cells for Repeated Deposition of Aerosolized Tobramycin.

As an alternative to technically demanding and ethically debatable animal models, the use of organotypic and disease-relevant human cell culture models may improve the throughput, speed, and success rate for the translation of novel anti-infectives into the clinic. Besides bacterial killing, host cell viability and barrier function appear as relevant but seldomly measured readouts. Moreover, bacterial virulence factors and signaling molecules are typically not addressed in current cell culture models. Here, we describe a reproducible protocol for cultivating barrier-forming human bronchial epithelial cell monolayers on Transwell inserts and infecting them with microclusters of pre-grown mature Pseudomonas aeruginosa PAO1 biofilms under the air-liquid interface conditions. Bacterial growth and quorum sensing molecules were determined upon tobramycin treatment. The host cell response was simultaneously assessed through cell viability, epithelial barrier function, and cytokine release. By repeated deposition of aerosolized tobramycin after 1, 24, and 48 h, bacterial growth was controlled (reduction from 10 to 4 log10 CFU/mL), which leads to epithelial cell survival for up to 72 h. E-cadherin's cell-cell adhesion protein expression was preserved with the consecutive treatment, and quorum sensing molecules were reduced. However, the bacteria could not be eradicated and epithelial barrier function was impaired, similar to the currently observed situation in the clinic in lack of more efficient anti-infective therapies. Such a human-based in vitro approach has the potential for the preclinical development of novel anti-infectives and nanoscale delivery systems for oral inhalation.

Electrothermoplasmonic Trapping and Dynamic Manipulation of Single Colloidal Nanodiamond.

Low-power trapping of nanoscale objects can be achieved by using the enhanced fields near plasmonic nanoantennas. Unfortunately, in this approach the trap site is limited to the position of the plasmonic hotspots and continuous dynamic manipulation is not feasible. Here, we report a low-frequency electrothermoplasmonic tweezer (LFET) that provides low-power, high-stability and continuous dynamic manipulation of a single nanodiamond. LFET harnesses the combined action of the laser illumination of a plasmonic nanopillar antenna array and low-frequency alternating current (ac) electric field to establish an electrohydrodynamic potential capable of the stable trapping and dynamic manipulation of single nanodiamonds. We experimentally demonstrate the fast transport, trapping, and dynamic manipulation of a single nanodiamond using a low-frequency ac field below 5 kHz and low-laser power of 1 mW. This nanotweezer platform for nanodiamond manipulation holds promise for the scalable assembly of single photon sources for quantum information processing and low noise quantum sensors.

A Custom-Made Device for Reproducibly Depositing Pre-metered Doses of Nebulized Drugs on Pulmonary Cells in vitro.

The deposition of pre-metered doses (i.e., defined before and not after exposition) at the air-liquid interface of viable pulmonary epithelial cells remains an important but challenging task for developing aerosol medicines. While some devices allow quantification of the deposited dose after or during the experiment, e.g., gravimetrically, there is still no generally accepted way to deposit small pre-metered doses of aerosolized drugs or pharmaceutical formulations, e.g., nanomedicines. Here, we describe a straightforward custom-made device, allowing connection to commercially available nebulizers with standard cell culture plates. Designed to tightly fit into the approximately 12-mm opening of either a 12-well Transwell® insert or a single 24-well plate, a defined dose of an aerosolized liquid can be directly deposited precisely and reproducibly (4.8% deviation) at the air-liquid interface (ALI) of pulmonary cell cultures. The deposited dose can be controlled by the volume of the nebulized solution, which may vary in a range from 20 to 200 µl. The entire nebulization-deposition maneuver is completed after 30 s and is spatially homogenous. After phosphate-buffered saline (PBS) deposition, the viability and barrier properties transepithelial electrical resistance (TEER) of human bronchial epithelial Calu-3 cells were not negatively affected. Straightforward in manufacture and use, the device enables reproducible deposition of metered doses of aerosolized drugs to study the interactions with pulmonary cell cultures grown at ALI conditions.

Tobramycin Liquid Crystal Nanoparticles Eradicate Cystic Fibrosis-Related Pseudomonas aeruginosa Biofilms.

Pseudomonas aeruginosa biofilms cause persistent and chronic infections, most known clinically in cystic fibrosis (CF). Tobramycin (TOB) is a standard anti-pseudomonal antibiotic; however, in biofilm infections, its efficacy severely decreases due to limited permeability across the biofilm matrix. Herewith, a biomimetic, nanostructured, lipid liquid crystal nanoparticle-(LCNP)-formulation is discovered to significantly enhance the efficacy of TOB and eradicate P. aeruginosa biofilm infections. Using an advanced, biologically-relevant co-culture model of human CF bronchial epithelial cells infected with P. aeruginosa biofilms at an air-liquid interface, nebulized TOB-LCNPs completely eradicated 1 × 109 CFU mL-1 of P. aeruginosa after two doses, a 100-fold improvement over the unformulated antibiotic. The enhanced activity of TOB is not observed with a liposomal formulation of TOB or with ciprofloxacin, an antibiotic that readily penetrates biofilms. It is demonstrated that the unique nanostructure of the LCNPs drives the enhanced penetration of TOB across the biofilm barrier, but not through the healthy lung epithelium barrier, significantly increasing the available antibiotic concentration at the site of infection. The LCNPs are an innovative strategy to improve the performance of TOB as a directed pulmonary therapy, enabling the administration of lower doses, reducing the toxicity, and amplifying the anti-biofilm activity of the anti-pseudomonal antibiotic.

A pulmonary mucus surrogate for investigating antibiotic permeation and activity against Pseudomonas aeruginosa biofilms.

BACKGROUND: Pulmonary infections associated with Pseudomonas aeruginosa can be life-threatening for patients suffering from chronic lung diseases such as cystic fibrosis. In this scenario, the formation of biofilms embedded in a mucus layer can limit the permeation and the activity of anti-infectives. OBJECTIVES: Native human pulmonary mucus can be isolated from endotracheal tubes, but this source is limited for large-scale testing. This study, therefore, aimed to evaluate a modified artificial sputum medium (ASMmod) with mucus-like viscoelastic properties as a surrogate for testing anti-infectives against P. aeruginosa biofilms. METHODS: Bacterial growth in conventional broth cultures was compared with that in ASMmod, and PAO1-GFP biofilms were imaged by confocal microscopy. Transport kinetics of three antibiotics, tobramycin, colistin, and ciprofloxacin, through native mucus and ASMmod were studied, and their activity against PAO1 biofilms grown in different media was assessed by determination of metabolic activity and cfu. RESULTS: PAO1(-GFP) cultured in human pulmonary mucus or ASMmod showed similarities in bacterial growth and biofilm morphology. A limited permeation of antibiotics through ASMmod was observed, indicating its strong barrier properties, which are comparable to those of native human mucus. Reduced susceptibility of PAO1 biofilms was observed in ASMmod compared with LB medium for tobramycin and colistin, but less for ciprofloxacin. CONCLUSIONS: These findings underline the importance of mucus as a biological barrier to antibiotics. ASMmod appears to be a valuable surrogate for studying mucus permeation of anti-infectives and their efficacy against PAO1 biofilms.

Disease Models: Lung Models for Testing Drugs Against Inflammation and Infection.

Lung diseases have increasingly attracted interest in the past years. The all-known fear of failing treatments against severe pulmonary infections and plans of the pharmaceutical industry to limit research on anti-infectives to a minimum due to cost reasons makes infections of the lung nowadays a "hot topic." Inhalable antibiotics show promising efficacy while limiting adverse systemic effects to a minimum. Moreover, in times of increased life expectancy in developed countries, the treatment of chronic maladies implicating inflammatory diseases, like bronchial asthma or chronic obstructive pulmonary disease, becomes more and more exigent and still lacks proper treatment.In this chapter, we address in vitro models as well as necessary in vivo models to help develop new drugs for the treatment of various severe pulmonary diseases with a strong focus on infectious diseases. By first presenting the essential hands-on techniques for the setup of in vitro models, we intend to combine these with already successful and interesting model approaches to serve as some guideline for the development of future models. The overall goal is to maximize time and cost-efficacy and to minimize attrition as well as animal trials when developing novel anti-infective therapeutics.