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Cost-effective bacterial detection system based on dynamic properties enhanced with nanoplasmonic biomarkers: a promising analytical approach with significantly reduced detection time.

We demonstrate sensitive electric field measurements by coherent homodyne amplification of electric-field induced second harmonic generation (E-FISH) technique. In the process of E-FISH, an applied electric field breaks centrosymmetryof an otherwise homogeneous medium, in turn promoting generation of second harmonic frequency of an incident field. Due to weak third-order hyperpolarizability and requirement of an applied field to break the symmetry, the E-FISH technique has been mainly used to study high fields, also requiring strong optical field and sensitive detection. Here we superimpose the E-FISH signal with an auxilliary beam,also termed a local oscillator (LO), at double of the incident frequency. Coherent superposition of the LO and the E-FISH output allows for a homodyne amplification of the otherwise weak nonlinear signal. We have demonstrated an increase of signal to noise by a factor of 7, which results in a measurement time reduction of a factor of 49. This technique, local oscillator E-FISH, LOE-FISH, has a number of advantages: Detection with intensified detectors is not required. Furthermore, instead of mJ pulsed lasers, we can work with μJ pulsed lasers, which allows measuring at repetition rates of MHz and opens single shot and real time capability. The LOE-FISH technique increases in sensitivity at lower electric field. Our work is a demonstration of the principle. Already with our first results from the demonstration one can see the high potential of LOE-FISH.

Fine-needle aspiration cytology is a minimally invasive diagnostic strategy. However, collected specimens have to undergo a series of complex preparation steps, thus limiting the use of fine-needle...

This article presents an optical platform for studying the dynamics of nanoparticle assisted pulsed laser optoporation of individual living cells. Here plasmonic nanoparticles (NPs) act as markers of the exact spatial position of living cell membranes and as an enhancer for localized pulsed laser perforation. High contrast NP imaging using reflected light microscopy (RLM) allows accurate and automatic laser targeting at individual NPs for spatially controlled laser optoporation of single cells at a single point. The NP imaging method is compatible with fluorescence microscopy and a cellular incubator that allows study of real-time perforation kinetics of live cells and the optomechanical interaction of NPs with membranes. These parameters are of great interest for the development and experimental implementation of the technology of pulsed laser optoporation and transfection applied to single living cells as well as to bulk-level assays.

We report on experimental observations of phenomenological self-trapping in plasmonic colloids of varying plasmon peaks in the visible/near infrared. A femtosecond (fs) oscillator is used in both pulsed (35 fs, 76 MHz) and continuous wave (cw) operation for comparison. We show that for both modes and for all examined colloids (and under typically applied external focusing conditions in self-trapping studies in colloidal media) nonlinear propagation is governed by thermal defocusing of the focused beam, which precedes the steady-state regime reached by particle diffusion, even far from the plasmon resonance (or equivalently for non-plasmonic colloids, even for low absorption coefficients). A strategy for the utilization of high repetition fs pulses to mitigate thermal lensing and promote gradient force-induced self-trapping is discussed. Notably, nonlinear thermal lensing is further accompanied by natural convection due to the horizontal configuration of the setup. Under resonant illumination, for both fs and cw cases, we observe mode break-up of the beam profile, most likely due to azimuthal modulation instability. Importantly, time-resolved observations of the break-up indicate that in the fs case, thermal convection heat transfer is reduced in magnitude and significantly decoupled in time from thermal conduction, presumably due to temperature increase confinement near the particles. We anticipate that our findings will trigger interest toward the use of high repetition fs pulses for self-channeling applications in nano-colloids.

We evaluate the threshold power for self-focusing in gold nanorod colloids of varying concentration by a power limiting method in the femtosecond filamentation regime. The pulses are tuned near the longitudinal plasmon peak of the nanorods, leading to saturation of linear absorption and reshaping of the particles. We evaluated the last two effects by optical transmission measurements and spectroscopic analysis and estimated that considerable particle deformation does not occur before the collapse of the beam. We performed numerical simulations based on the experimental results, and evaluated only a subtle, monotonically increasing enhancement of the nonlinear refractive index of the host material (water) as the nanoparticles concentration increases. The role of higher-order contributions is discussed. Our work provides an alternative characterization approach of ultrafast nonlinearities in absorbing media. It further emphasizes that self-focusing of intense femtosecond pulses in gold nanocomposites is hampered by the ultrafast modulation of the susceptibility of the metal.

Reliable cytopathological diagnosis requires new methods and approaches for the rapid and accurate determination of all cell types. This is especially important when the number of cells is limited, such as in the cytological samples of fine-needle biopsy. Immunoplasmonic-multiplexed- labeling may be one of the emerging solutions to such problems. However, to be accepted and used by the practicing pathologists, new methods must be compatible and complementary with existing cytopathology approaches where counterstaining is central to the correct interpretation of immunolabeling. In addition, the optical detection and imaging setup for immunoplasmonic-multiplexed-labeling must be implemented on the same cytopathological microscope, not interfere with standard H&amp, E imaging, and operate as a second easy-to-use imaging method. In this article, we present multiplex imaging of four types of nanoplasmonic markers on two types of H&amp, E-stained cytological specimens (formalin-fixed paraffin embedded and non-embedded adherent cancer cells) using a specially designed adapter for SI dark-field microscopy. The obtained results confirm the effectiveness of the proposed optical method for quantitative and multiplex identification of various plasmonic NPs, and the possibility of using immunoplasmonic-multiplexed-labeling for cytopathological diagnostics.

The rapid advances of genetic and genomic technology indicate promising therapeutic potential of genetic materials for regulating abnormal gene expressions causing diseases and disorders. However, targeted intracellular delivery of RNA therapeutics still remains a major challenge hindering the clinical translation. In this study, an elaborated plasmonic optoporation approach is proposed to efficiently and selectively transfect specific cells. The site-specific optoporation is obtained by tuning the spectral range of a supercontinuum pulsed picosecond laser in order for each individual cell binding gold nanostar with their unique resonance peak to magnify the local field strength in the near-infrared region and facilitate a selective delivery of small interfering RNA, messenger RNA, and Cas9-ribonucleoprotein into human retinal pigment epithelial cells. Numerical simulations indicate that optoporation is not due to a plasma-mediated process but rather due to a highly localized temperature rise both in time (few nanoseconds) and space (few nanometers). Taking advantage of the numerical simulation and fine-tuning of the optical strategy, the perforated lipid bilayer of targeted cells undergoes a membrane recovery process, important to retain their viability. The results signify the prospects of antibody functionalized nanostar-mediated optoporation as a simple and realistic gene delivery approach for future clinical practices.

Porous alloy nanomaterials are important for applications in catalysis, sensing, and actuation. Chemical and electrochemical etching are two methods to form porous nanostructures by dealloying bimetallic nanoparticles (NPs). However, it is not clear how the NPs evolve during these etching processes. Insight into the morphological and compositional transformations of the NPs during the etching is critical to understanding the nanoscale details of the dealloying process. Here, using in situ liquid phase transmission electron microscopy, the structural evolution of individual AuAg alloy NPs is tracked during both chemical and electrochemical etching of their Ag component. The observations show that the electrochemical etching produces NPs with more uniform pore sizes than the chemical etching and enables tuning the NPs porosity by modulating the electrochemical potential. The results show that at the initial stages of both etching methods, Au-rich passivation layer forms on the surface of the NPs, which is critical in preserving the NP's porous shell as pores form underneath this layer during the etching. These findings describing the selective etching and dealloying of AuAg NPs provide a critical insight needed to control the morphology and composition of porous multimetallic NPs, and paves the way for synthesizing nanomaterials with tailored chemical and physical properties for various applications.

Deep image clustering approaches typically use autoencoder architectures to learn compressed latent representations suitable for clustering tasks. However, they do not effectively regulate the latent space during training, leading to low performance and diminished applicability to different datasets. In this paper, we propose a deep clustering model combining maximum mean discrepancy (MMD) regularization and self-learning clustering optimization to mitigate this problem. Specifically, we first train the network to improve its image reconstruction ability by minimizing both reconstruction loss and MMD divergence from a target distribution. Then, the model gradually learns from its own high-confidence predictions to further optimize the latent distribution. We validate the network’s performance on different benchmark image sets using standard clustering metrics, without changing network configuration or adjusting hyper-parameters between datasets. The proposed model provides top clustering performance across datasets while being more robust than state-of-the-art methods.

Automatic identification of subcellular compartments of proteins in fluorescence microscopy images is an important task to quantitatively evaluate cellular processes. A common problem for the development of deep learning based classifiers is that there is only a limited number of labeled images available for training. To address this challenge, we propose a new approach for subcellular organelles classification combining an effective and efficient architecture based on a compact Convolutional Neural Network and deep embedded clustering algorithm. We validate our approach on a benchmark of HeLa cell microscopy images. The network both yields high accuracy that outperforms state of the art methods and has significantly small number of parameters. More interestingly, experimental results show that our method is strongly robust against limited labeled data for training, requiring four times less annotated data than usual while maintaining the high accuracy of 93.9%.

A parametric investigation is conducted to determine favored threshold conditions for self-focusing under resonant interactions in different plasmonic nano-colloids. The theory predicts lower threshold intensities for anisotropic particles.

Plasmonic nanostructures have raised the interest of biomedical applications of surface-enhanced Raman scattering (SERS). To improve the enhancement and produce sensitive SERS probes, porous Au-Ag alloy nanoparticles (NPs) are synthesized by dealloying Au-Ag alloy NP-precursors with Au or Ag core in aqueous colloidal environment through galvanic replacement reaction. The novel designed core-shell Au-Ag alloy NP-precursors facilitate controllable synthesis of porous nanostructure, and dealloying degree during the reaction has significant effect on structural and spectral properties of dealloyed porous NPs. Narrow-dispersed dealloyed NPs are obtained using NPs of Au/Ag ratio from 10/90 to 40/60 with Au and Ag core to produce solid core@porous shell and porous nanoshells, having rough surface, hollowness, and porosity around 30-60%. The clean nanostructure from colloidal synthesis exhibits a redshifted plasmon peak up to near-infrared region, and the large accessible surface induces highly localized surface plasmon resonance and generates robust SERS activity. Thus, the porous NPs produce intensely enhanced Raman signal up to 68-fold higher than 100 nm AuNP enhancement at single-particle level, and the estimated Raman enhancement around 7800, showing the potential for highly sensitive SERS probes. The single-particle SERS probes are effectively demonstrated in quantitative monitoring of anticancer drug Doxorubicin release.

In this article, we present a multiperiodic nanohole array structure for improved sensing. The structure consists a series of rows of nanoholes, each having a different period in an ascending order. A monochromatic source illuminates the structure, and a resonance condition is met for the row having a momentum matching Bloch wave, which leads to extraordinary optical transmission. With this new plasmonic structure, the sensing signal can be retrieved using the spatial position of the transmission maxima. This setup requires a simple optical setup while achieving increased resolution and accuracy. A resolution of 4.6×10−6 refractive index units is achieved, which is comparable to surface plasmon resonance system based on the Kretchmann configuration.

Vision loss caused by retinal diseases affects hundreds of millions of individuals worldwide. The retina is a delicate central nervous system tissue stratified into layers of cells with distinct roles. Currently, there is a void in treatments that selectively target diseased retinal cells, and current therapeutic paradigms present complications associated with off-target effects. Herein, as a proof of concept, we introduce an in vivo method using a femtosecond laser to locally optoporate retinal ganglion cells (RGCs) targeted with functionalized gold nanoparticles (AuNPs). We provide evidence that AuNPs functionalized with an antibody toward the cell-surface voltage-gated K+ channel subunit KV1.1 can selectively deliver fluorescently tagged siRNAs or fluorescein isothiocyanate-dextran dye into retinal cells when irradiated with an 800 nm 100 fs laser. Importantly, neither AuNP administration nor irradiation resulted in RGC death. This system provides a novel, non-viral-based approach that has the potential to selectively target retinal cells in diseased regions while sparing healthy areas and may be harnessed in future cell-specific therapies for retinal degenerative diseases.

Gold–silver alloy core silica shell nanoparticles (Au/Ag@SiO2 NPs) with fully accordable photophysical properties were developed and used as contrast agent for multiplexed cell-imaging applications. Using a seed growth strategy, these nanoparticular labels can be designed to suit specific experimental needs and be clearly identified based on their distinctive combination of scattering and fluorescence colors. In this Article, the multiplexed cell-imaging capabilities of Au/Ag@SiO2 NPs are presented using four different combinations of core composition and fluorescence colors. In a proof of concept experiment, 86% of total nanoparticles were correctly identified by an optical microscope with a system offering fluorescence and darkfield detection capabilities. This multiplatform identification strategy was successfully applied for the detection of four different nanoparticular architectures in cell-imaging experiments.

Direct microscopy interpretation of fine-needle biopsy cytological samples is routinely used by practicing cytopathologists. Adding possibility to identify selective and multiplexed biomarkers on the same samples and with the same microscopy technique can greatly improve diagnostic accuracy. In this article, we propose to use biomarkers based on designable plasmonic nanoparticles (NPs) with unique optical properties and excellent chemical stability that can satisfy the above-mentioned requirements. By finely controlling the size and composition of gold-silver alloy NPs and gold nanorods, the NPs plasmonic resonance properties, such as scattering efficiency and resonance peak spectral position, are adjusted in order to provide reliable identification and chromatic differentiation by conventional direct microscopy. Efficient darkfield NPs imaging is performed by using a novel circular side illumination adaptor that can be easily integrated into any microscopy setup while preserving standard cytopathology visualization method. The efficiency of the proposed technology for fast visual detection and differentiation of three spectrally distinct NP-markers is demonstrated in different working media, thus confirming the potential application in conventional cytology preparations. It is worth emphasizing that the presented technology does not interfere with standard visualization with immunohistochemical staining, but should rather be considered as a second imaging modality to confirm the diagnostics.

Retinoblastoma (RB) is a rare form of cancer of the retina most prevalent in young children. We successfully show that laser-induced cell disruption, mediated by gold plasmonic nanoparticle (NP), is a potential and efficient therapy to kill the cancerous cells. The proof of concept is demonstrated in vitro on cultured Y79 RB cancer cells with a nanosecond laser at 527 nm, for both attached cells at the bottom of a Petri dish and for floating, clustered cells in a viscous vitreous phantom comprised of hyaluronan. We report a cellular death of 82% after irradiation in classic culture medium and a cellular death of 98% in vitreous phantom, for similar number of NPs in each sample. It is found that the NPs efficiently penetrate the floating Y79 clusters cells in the vitreous phantom, leading to a cellular death of over 85% even within the centre of the aggregates. The proposed treatment technique is based on a similar nanosecond laser used to eliminate floaters in the vitreous, but with much lower (100-1000 times) fluences of 20 J cm-2 .

There is a current void in efficient, cell-specific, retinal drug delivery systems, thus developing a safe, effective, selective drug delivery system would open novel therapeutic avenues. We previously demonstrated that femtosecond (fs) laser irradiation can selectively transfect DNA plasmids into cultured cells in the presence of functionalised gold nanoparticles (AuNPs) (1). Here, we sought out to selectively optoporate retinal cells in vivo with functionalized AuNPs and a 800nm fs laser. The cell-surface Kv1.1 voltage-gated channel was chosen to target retinal ganglion cells (RGCs) in the rat retina. The eyes of anesthetized rats were placed in the beam path of an optical system consisting of a fs laser and an ophthalmoscope for fundus visualization. Following Kv1.1-AuNP and FITC-dextran intravitreal injection and incubation, irradiation resulted in FITC uptake by retinal cells. In addition, similar experiments with Cy3-siRNA clearly show that the technique can effectively deliver siRNA into RGCs. Importantly, neither AuNP intravitreal injection nor irradiation resulted in RGC death, as determined by RBPMS quantification 1 week following AuNP injection and/or irradiation. Since living biological tissues absorb energy very weakly at 800nm, this non-invasive tool may provide a safe, cost effective approach to selectively target retinal cells and limit complications associated with surgical interventions, and potential biological hazards associated with viral-based gene therapy. In addition, given the extensive use of lasers in ophthalmic practice, our proposed technology may be seamlessly inserted to current clinical setups. (1) E. Bergeron et al, Nanoscale, 7, 17836 (2015).

Intracellular delivery of molecular cargo is the basis for a plethora of therapeutic applications, including gene therapy and cancer treatment. An efficient method to perform intracellular delivery is the photoactivation of nanomaterials that bear releasable molecular cargo. However, potential in vivo applications of this method are limited by our ability to deliver nanomaterials and light in tissue. In this paper, we will present method to perform intracellular delivery of molecular cargo on live cells and tissue by using a reusable, needle-like optofluidic probe capable of penetrating soft tissue. The probe consists of a dual-core glass fiber, enabling simultaneous light and liquid delivery. First, we used the optofluidic probe to confine an intracellular delivery mixture, composed of 100 nm gold nanoparticles (AuNP) and membrane-impermeable calcein, in the vicinity of cancer cells and mouse retinal explants. Secondly, we delivered nanosecond (ns) laser pulses (wavelength: 532 nm; duration: 5 ns; 30-90 mJ/cm2) using the same probe and without introducing a AuNP cells incubation step. We found that AuNP photo-activation caused localized and reversible disruption of the cell membrane, enabling calcein delivery into the cytoplasm. We measured 67% intracellular delivery efficacy and showed that the optofluidic probe can be used to treat cells with single-cell precision. The method presented here can facilitate in vivo treatments in soft tissue of small animals (e.g. brain, retina), such us nanomaterial-assisted neuro-stimulation, transfection and tumor elimination.

Irradiating metallic nanostructures by an ultrafast laser beam produces highly localised processes on the nanoscale in the surrounding medium. This particular process is mainly attributed to the surface plasmon resonance of the nanostructures. When these nanomaterials are colloidal nanoparticles, their irradiation by an ultrafast laser could results in a highly localized plasma, heat production, pressure wave in the liquid and finally nanocavitation around the nanoparticles (1-5) . In this invited talk, I will cover recent developments of fundamentals aspects describing multiscale processes on the nanoscale as well as on ultrashort time (femtosecond to nanosecond). Multiphysics models based on electromagnetism, two-temperature model, thermodynamics and fluid dynamics have been developed to describe theoretically the productions of nanoplasma, pressure waves and nanocavitation. Models were successfully compared with experiments including shadowgraphy showing nanobubble dynamics and an all-optical pump-probe technique enabling the detection of plasmonic enhanced nanocavitation and pressure wave generation. Applications are in nanomedicine (6-7) in which ultrafast laser irradiation of these nanoparticles located close to a living cell, can produce a nanosurgery, can stimulate neurons or induce drug delivery. (1) E. Boulais et al, Nanoletters, 12, 4763-4769 (2012); (2) E. Boulais et al, Journal of Photochemistry and Photobiology C, 17, 26-49 (2013); (3) R. Lachaine et al ACS Photonics, 1, 331-336 (2014); (4) A. Dagallier, et al Nanoscale 9 (9), 3023-3032, (2017); (5) V. T. De Lille et al, to be submitted (2018); (6) E. Bergeron et al, Nanoscale, 7, 17836-17847 (2015); (7) F. Lavoie-Cardinal et al Scientific Reports, 6, 20619 (2016).

We present the development of an innovative technology for quantitative multiplexed cytology analysis based on the application of spectrally distinctive plasmonic nanoparticles (NPs) as optical probes and on cost-effective side-illumination multispectral darkfield microscopy (SIM) as the differential NP imaging method. SIM is based on lateral illumination by arrays of discrete color RGB light emitting diodes (LEDs) of spectrally adjusted plasmonic NPs and consecutive detection by the conventional CMOS color camera. We demonstrate the enhanced contrast and higher resolution of our method for individual NP detection in the liquid medium and of NP markers attached on the cell membrane in a cytology preparation by comparing it to the conventional darkfield microscopy (DFM). The proposed illumination and detection system is compatible with current clinical microscopy equipment used by pathologists and can greatly simplify the adaptation of plasmonic NPs as novel reliable and stable biological multiplexed chromatic markers for biodetection and diagnosis.

A semi-analytical formalism for the optical properties of a metal-insulator-metal periodic nanostructure using coupled-mode theory is presented. This structure consists in a dielectric layer in between two metallic layers with periodic one-dimensional nanoslit corrugation. The model is developed using multiple-scattering formalism, which defines transmission and reflection coefficients for each of the interface as a semi-infinite medium. Total transmission is then calculated using a summation of the multiple paths of light inside the structure. This method allows finding an exact solution for the transmission problem in every dimension regime, as long as a sufficient number of diffraction orders and guided modes are considered for the structure. The resonant modes of the structure are found to be related to the metallic slab only and to a combination of both the metallic slab and dielectric layer. This model also allows describing the resonant behavior of the system in the limit of a small dielectric layer, for which discontinuities in the dispersion curves are found. These discontinuities result from the out-of-phase interference of the different diffraction orders of the system, which account for field interaction for both inner interfaces of the structure.

Photons interaction with metallic nanoparticles can excite a resonant plasmon that concentrates energy at the nanoscale. At high intensity, this quasi-particle decays into a photoexcited nanoplasma that triggers the generation of nanobubbles, which can be used for imaging and therapeutic purposes. This highly nonlinear wavelength-dependent process is controlled by the nanoparticle material, shape, and size in intricate ways, which justifies the need for a systematic design approach that currently lacks in the field. To palliate to this, we developed in this work a computational framework that enables the efficient in silico screening of large libraries of spherically symmetric structures and metallic materials. Using this framework, we have investigated the nanocavitation performance of spherical nanoparticles with more than 14 million combinations of materials, shapes, sizes, and irradiation conditions, from which we could distill general principles for the design of durable nanoantennas. In the near-inf...

Controlling the interaction between linking components and nanoobjects is expected to offer exciting new opportunities to induce tunable assembly and modulate the topology and properties of nanostructures. In this work, engineering the exclusive and inclusive host–guest interaction between cucurbit[n]urils and salts is demonstrated as a novel strategy to simultaneously realize tunable assembly of plasmonic nanoparticles (NPs) with controllable size and composition and manipulate the stability of the assemblies. As a proof of concept, one-dimensional nanochains of gold, silver, and their alloys with rigid subnanometer interparticle separation and widely tunable optical properties are generated. These exhibit strong dipole coupling between the plasmonic NPs, confirmed by experiments and theoretical simulations. This controllable assembly principle will lead to significant interest not only in supramolecular chemistry and the interactions between supramolecular and plasmonic NPs but also in interaction mecha...

Metallic nanoparticles are routinely used as nanoscale antenna capable of absorbing and converting photon energy with subwavelength resolution. Many applications, notably in nanomedicine and nanobiotechnology, benefit from the enhanced optical properties of these materials, which can be exploited to image, damage, or destroy targeted cells and subcellular structures with unprecedented precision. Modern inorganic chemistry enables the synthesis of a large library of nanoparticles with an increasing variety of shapes, composition, and optical characteristic. However, identifying and tailoring nanoparticles morphology to specific applications remains challenging and limits the development of efficient nanoplasmonic technologies. In this work, we report a strategy for the rational design of gold plasmonic nanoshells (AuNS) for the efficient ultrafast laser-based nanoscale bubble generation and cell membrane perforation, which constitute one of the most crucial challenges toward the development of effective gene therapy treatments. We design an in silico rational design framework that we use to tune AuNS morphology to simultaneously optimize for the reduction of the cavitation threshold while preserving the particle structural integrity. Our optimization procedure yields optimal AuNS that are slightly detuned compared to their plasmonic resonance conditions with an optical breakdown threshold 30% lower than randomly selected AuNS and 13% lower compared to similarly optimized gold nanoparticles (AuNP). This design strategy is validated using time-resolved bubble spectroscopy, shadowgraphy imaging and electron microscopy that confirm the particle structural integrity and a reduction of 51% of the cavitation threshold relative to optimal AuNP. Rationally designed AuNS are finally used to perforate cancer cells with an efficiency of 61%, using 33% less energy compared to AuNP, which demonstrate that our rational design framework is readily transferable to a cell environment. The methodology developed here thus provides a general strategy for the systematic design of nanoparticles for nanomedical applications and should be broadly applicable to bioimaging and cell nanosurgery.

Surface plasmon resonance (SPR) spectroscopy has been used for the detection of reversible refractive index changes that occur in a redox mediator film during potentiostatic oxydoreduction. We investigated the theoretical and experimental parameters of electrochemical SPR (eSPR) sensing using as an example of the electroactive labels the Methylene Blue (MB) organic dye. The optimal eSPR interrogation method for MB sensing in a solution and in a uniform interfacial thin film formed by stem-loop oligonucleotides tagged with MB are evaluated. Electrochemical activation of the optical SPR response depends on the local MB concentration and can be used to design sensitive and highly selective biosensing approaches.

Precise spatial and temporal control of pressure stimulation at the nanometer scale is essential for the fabrication and manipulation of nano-objects, and for exploring single-molecule behaviour of matter under extreme conditions. However, state-of-the-art nano-mechanical transducers require sophisticated driving hardware and are currently limited to moderate pressure regimes. Here we report a gold plasmonic bowtie (AuBT) nano-antennas array that can generate extreme pressure stimulus of ∼100 GPa in the ps (10−12 s) time scale with sub-wavelength resolution upon irradiation with ultra-short laser pulses. Our method leverages the non-linear interaction of photons with water molecules to excite a nano-plasma in the plasmon-enhanced near-field and induce extreme thermodynamic states. The proposed method utilizes laser pulses, which in contrast to micro- and nano-mechanical actuators offers simplicity and versatility. We present time-resolved shadowgraphic imaging, electron microscopy and simulation data that suggest that our platform can efficiently create cavitation nano-bubbles and generate intense pressure in specific patterns, which can be controlled by the selective excitation of plasmon modes of distinct polarizations. This novel platform should enable probing non-invasively the mechanical response of cells and single-molecules at time and pressure regimes that are currently difficult to reach with other methods.

Intracellular delivery of molecular cargo is the basis for a plethora of therapeutic applications, including gene therapy and cancer treatment. A very efficient method to perform intracellular delivery is the photo-activation of nanomaterials that have been previously directed to the cell vicinity and bear releasable molecular cargo. However, potential in vivo applications of this method are limited by our ability to deliver nanomaterials and light in tissue. Here, we demonstrate intracelullar delivery using a needle-like optofluidic probe capable of penetrating soft tissue. Firstly, we used the optofluidic probe to confine an intracellular delivery mixture, composed of 100 nm gold nanoparticles (AuNP) and membrane-impermeable calcein, in the vicinity of cancer cells. Secondly, we delivered nanosecond (ns) laser pulses (wavelength: 532 nm; duration: 5 ns) using the same probe and without introducing a AuNP cells incubation step. The AuNP photo-activation caused localized and reversible disruption of the cell membrane, enabling calcein delivery into the cytoplasm. We measured 67% intracellular delivery efficacy and showed that the optofluidic probe can be used to treat cells with single-cell precision. Finally, we demonstrated targeted delivery in tissue (mouse retinal explant) ex vivo. We expect that this method can enable nanomaterial-assisted intracellular delivery applications in soft tissue (e.g. brain, retina) of small animals.

Vision loss caused by retinal diseases affects hundreds of millions of individuals worldwide. The retina is a delicate central nervous system tissue stratified into layers of cells with distinct roles. Currently, there is a void in treatments that selectively target diseased retinal cells, and current therapeutic paradigms present complications associated with off-target effects. Herein, as a proof of concept, we introduce an in vivo method using a femtosecond laser to locally optoporate retinal ganglion cells (RGCs) targeted with functionalized gold nanoparticles (AuNPs). We provide evidence that AuNPs functionalized with an antibody toward the cell-surface voltage-gated K

Obesity gives rise to metabolic complications by mechanisms that are poorly understood. Although chronic inflammatory signaling in adipose tissue is typically associated with metabolic deficiencies linked to excessive weight gain, we identified a subset of neuropilin-1 (NRP1)-expressing myeloid cells that accumulate in adipose tissue and protect against obesity and metabolic syndrome. Ablation of NRP1 in macrophages compromised lipid uptake in these cells, which reduced substrates for fatty acid β-oxidation and shifted energy metabolism of these macrophages toward a more inflammatory glycolytic metabolism. Conditional deletion of NRP1 in LysM Cre-expressing cells leads to inadequate adipose vascularization, accelerated weight gain, and reduced insulin sensitivity even independent of weight gain. Transfer of NRP1+ hematopoietic cells improved glucose homeostasis, resulting in the reversal of a prediabetic phenotype. Our findings suggest a pivotal role for adipose tissue-resident NRP1+-expressing macrophages in driving healthy weight gain and maintaining glucose tolerance.

Sn-containing Si and Ge (Ge1-y-xSixSny) alloys are an emerging family of semiconductors with the potential to impact group IV material-based devices. These semiconductors provide the ability to independently engineer both the lattice parameter and bandgap, which holds the premise to develop enhanced or novel photonic and electronic devices. With this perspective, we present detailed investigations of the influence of Ge1-y-xSixSny layers on the optical properties of Si and Ge based heterostructures and nanowires. We found that by adding a thin Ge1-y-xSixSny capping layer on Si or Ge greatly enhances light absorption especially in the near infrared range, leading to an increase in short-circuit current density. For the Ge1-y-xSixSny structure at thicknesses below 30 nm, a 14-fold increase in the short-circuit current is observed with respect to bare Si. This enhancement decreases by reducing the capping layer thickness. Conversely, decreasing the shell thickness was found to improve the short-circuit current in Si/Ge1-y-xSixSny and Ge/Ge1-y-xSixSny core/shell nanowires. The optical absorption becomes very important by increasing the Sn content. Moreover, by exploiting an optical antenna effect, these nanowires show extreme light absorption, reaching an enhancement factor, with respect to Si or Ge nanowires, on the order of 104 in Si/Ge0.84Si0.04Sn0.12 and 12 in Ge/Ge0.84Si0.04Sn0.12. Furthermore, we analyzed the optical response after the addition of a dielectric layer of Si3N4 to the Si/Ge1-y-xSixSny core-shell nanowire and found approximatively a 50% increase in the short-circuit current density for a dielectric layer of thickness equal to 45 nm and both a core radius and a shell thickness greater than 40 nm. The core−shell optical antenna benefits from a multiplication of enhancements contributed by leaky mode resonances in the semiconductor part and antireflection effects in the dielectric part.

In this article, the implementation of electrochemical plasmonic nanostructures functionalized with DNA-based structure-switching sensors is presented. eNanoSPR devices with open and microfluidic measurement cells are developed on the base of nanohole arrays in 100 nm gold film and applied for combined microscopic and electrochemical surface plasmon (eSPR) visualization. eSPR voltammograms and spectroscopy are performed using planar three electrode schematic with plasmonic nanostructure operated as working electrode. Limit of detection of eNanoSPR devices for oligonucleotide hybridization is estimated in the low nanomolar and applications for structure-switching electro-plasmonic sensing in complex liquids are discussed.