AbstractTwo-dimensional transition metal dichalcogenides, particularly MoS2, are interesting materials for many applications in aerospace research, radiation therapy and bioscience more in general. Since in many of these applications MoS2-based nanomaterials can be placed in an aqueous environment while exposed to ionizing radiation, both experimental and theoretical studies of their behaviour under these conditions is particularly interesting. Here, we study the effects of tiny imparted doses of 511 keV photons to MoS2 nanoflakes in water solution. To the best of our knowledge, this is the first study in which ionizing radiation on 2D-MoS2 occurs in water. Interestingly, we find that, in addition to the direct interaction between high-energy photons and nanoflakes, reactive chemical species, generated by γ-photons induced radiolysis of water, come into play a relevant role. A radiation transport Monte Carlo simulation allowed determining the elements driving the morphological and spectroscopical changes of 2D-MoS2, experimentally monitored by SEM microscopy, DLS, Raman and UV–vis spectroscopy, AFM, and X-ray photoelectron techniques. Our study demonstrates that radiolysis products affect the Molybdenum oxidation state, which is massively changed from the stable + 4 and + 6 states into the rarer and more unstable + 5. These findings will be relevant for radiation-based therapies and diagnostics in patients that are assuming drugs or contrast agents containing 2D-MoS2 and for aerospace biomedical applications of 2DMs investigating their actions into living organisms on space station or satellites.

Food allergies are an exceptional response of the immune system caused by the ingestion of specific foods. The main foods responsible for allergic reactions are milk, eggs, seafood, soy, peanuts, tree nuts, wheat, and their derived products. Chicken egg ovalbumin (OVA), a common allergen molecule, is often used for the clarification process of wine. Traces of OVA remain in the wine during the fining process, and they can cause significant allergic reactions in sensitive consumers. Consequently, the European Food Safety Authority (EFSA) and the American Food and Drug Administration (FDA) have shown the risks for allergic people to assume allergenic foods and food ingredients, including eggs. Commonly, OVA detection requires sophisticated and time-consuming analytical techniques. Intending to develop a faster assay, we designed a proof-of-concept non-Faradaic impedimetric immunosensor for monitoring the presence of OVA in wine. Polyclonal antibodies anti-OVA were covalently immobilised onto an 11-mercaptoundecanoic-acid (11-MUA)-modified gold surface. The developed immunosensor was able to detect OVA in diluted white wine without the need for an external probe or any pre-treatment step with a sensitivity of 0.20 µg/mL, complying with the limit established by the resolution OIV/COMEX 502–2012 for the quantification of allergens in wine.

Surface-enhanced Raman scattering (SERS) is of growing interest for a wide range of applications, especially for biomedical analysis, thanks to its sensitivity, specificity, and multiplexing capabilities. A crucial role for successful applications of SERS is played by the development of reproducible, efficient, and facile procedures for the fabrication of metal nanostructures (SERS substrates). Even more challenging is to extend the fabrication techniques of plasmonic nano-textures to atomic force microscope (AFM) probes to carry out tip-enhanced Raman spectroscopy (TERS) experiments, in which spatial resolution below the diffraction limit is added to the peculiarities of SERS. In this short review, we describe recent studies performed by our group during the last ten years in which novel nanofabrication techniques have been successfully applied to SERS and TERS experiments for studying bio-systems and molecular species of environmental interest.

The development of sensitive methods for the detection of endotoxin molecules, such as lipopolysaccharides (LPS), is essential for food safety and health control. Conventional analytical methods used for LPS detection are based on the pyrogen test, plating and culture-based methods, and the limulus amoebocyte lysate method (LAL). Alternatively, the development of reliable biosensors for LPS detection would be highly desirable to solve some critical issues, such as high cost and a long turnaround time. In this work, we present a label-free Surface-Enhanced Raman Spectroscopy (SERS)-based method for LPS detection in its free form. The proposed method combines the benefits of plasmonic enhancement with the selectivity provided by a specific anti-lipid A antibody (Ab). A high-enhancing nanostructured silver substrate was coated with Ab. The presence of LPS was quantitatively monitored by analyzing the changes in the Ab spectra obtained in the absence and presence of LPS. A limit of detection (LOD) and quantification (LOQ) of 12 ng/mL and 41 ng/mL were estimated, respectively. Importantly, the proposed technology could be easily expanded for the determination of other biological macromolecules.

Nanoparticles (NPs) coated with hyaluronic acid (HA) seem to be increasingly promising for targeted therapy due to HA chemical versatility, which allows them to bind drugs of different natures, and their affinity with the transmembrane receptor CD-44, overexpressed in tumor cells. However, an essential aspect for clinical use of NPs is formulation stability over time. For these reasons, analytical techniques capable of characterizing their physico-chemical properties are needed. In this work, poly(lactide-co-glycolide) (PLGA) NPs with an average diameter of 100-150 nm, coated with a few 10 s of nm of HA, were synthesized. For stability characterization, two complementary investigative techniques were used: Dynamic Light Scattering (DLS) and Surface-Enhanced Raman Scattering (SERS) spectroscopy. The first technique provided information on size, polidispersity index, and zeta-potential, and the second provided a deeper insight on the NP surface chemicals, allowing distinguishing of HA-coated NPs from uncoated ones. Furthermore, in order to estimate formulation stability over time, NPs were measured and monitored for two weeks. SERS results showed a progressive decrease in the signal associated with HA, which, however, is not detectable by the DLS measurements.

Controlling light at the nanoscale, the fascinating goal of nanophononics, allows to mold the flow of light in previously unknown ways, providing access to the study of the interaction of single molecules with confined optical fields. One of the most powerful spectroscopic approach to nanoscale is Tip-Enhanced Raman Spectroscopy (TERS). TERS combines the fingerprint character of Raman spectroscopy with the high spatial resolution of scanning probe microscopies (SPM), by taking advantage of plasmonic nanostructures placed at the apex of a SPM probe. Thanks to the excitation of Localized Surface Plasmon Resonances (LSPR), such system behaves as a nano-antenna, able to explore surfaces with nanometric spatial resolution and sensitivity up to single molecule level. Herein, we report on the fabrication and characterization of newly conceived nano-antennas. In particular, we illustrate a top-down approach for TERS tip fabrication, based on the solid-state dewetting of metal-sputtered Atomic-Force Microscopy probes. Such approach produces AFM-TERS tips, exhibiting high reproducibility and efficiency.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants that are typically released into the environment during the incomplete combustion of fossil fuels. Due to their relevant carcinogenicity, mutagenicity, and teratogenicity, it is urgent to develop sensitive and cost-effective strategies for monitoring them, especially in aqueous environments. Surface-enhanced Raman spectroscopy (SERS) can potentially be used as a reliable approach for this purpose, as it constitutes a valid alternative to traditional techniques, such as liquid and gas chromatography. Nevertheless, the development of an SERS-based platform for detection PAHs has so far been hindered by the poor adsorption of PAHs onto silver- and gold-based SERS-active substrates. To overcome this limitation, several research efforts have been directed towards the development of functionalized SERS substrates for the improvement of PAH adsorption. However, these strategies suffer from the interference that functionalizing molecules can produce in SERS detection. Herein, we demonstrate the feasibility of label-free detection of pyrene by using a highly porous 3D-SERS substrate produced by an inductively coupled plasma (ICP). Thanks to the coral-like nanopattern exhibited by our substrate, clear signals ascribable to pyrene molecules can be observed with a limit of detection of 23 nM. The observed performance can be attributed to the nanoporous character of our substrate, which combines a high density of hotspots and a certain capability of trapping molecules and favoring their adhesion to the Ag nanopattern. The obtained results demonstrate the potential of our substrates as a large-area, label-free SERS-based platform for chemical sensing and environmental control applications.

Two-dimensional nanomaterials, such as MoS2 nanosheets, have been attracting increasing attention in cancer diagnosis and treatment, thanks to their peculiar physical and chemical properties. Although the mechanisms which regulate the interaction between these nanomaterials and cells are not yet completely understood, many studies have proved their efficient use in the photothermal treatment of cancer, and the response to MoS2 nanosheets at the single-cell level is less investigated. Clearly, this information can help in shedding light on the subtle cellular mechanisms ruling the interaction of this 2D material with cells and, eventually, to its cytotoxicity. In this study, we use confocal micro-Raman spectroscopy to reconstruct the thermal map of single cells targeted with MoS2 under continuous laser irradiation. The experiment is performed by analyzing the water O-H stretching band around 3,400 cm−1 whose tetrahedral structure is sensitive to the molecular environment and temperature. Compared to fluorescence-based approaches, this Raman-based strategy for temperature measurement does not suffer fluorophore instability, which can be significant under continuous laser irradiation. We demonstrate that irradiation of human breast cancer MCF7 cells targeted with MoS2 nanosheets causes a relevant photothermal effect, which is particularly high in the presence of MoS2 nanosheet aggregates. Laser-induced heating is strongly localized near such particles which, in turn, tend to accumulate near the cytoplasmic membrane. Globally, our experimental outcomes are expected to be important for tuning the nanosheet fabrication process.

Polycyclic aromatic hydrocarbons (PAHs) are persistent organic pollutants mainly originating from the incomplete combustion of fossil fuels. Health and cancer risk arising from these compounds makes it quite urgent to monitor them in air, soil, and water. Since their concentrations in the environment are usually low (order of ng/L in water), highly sensitive techniques are required. Thanks to the large amount of structural information that it can offer, Surface Enhanced Raman Spectroscopy (SERS) has proven to be an effective alternative to traditional chromatographic techniques for the detection of PAHs. By exploiting the plasmonic properties of metallic nanostructures, this technique allows the detection of very low analyte concentration (nano-and pico-molar range), achieving even the limit of single-molecule detection. However, due to the ineffective adsorption of PAHs onto metallic SERS-active substrates, these surfaces are generally functionalized to increase their adhesion. Such chemical modifications can surely improve the selectivity of selected compounds but might hinder the effective direct contact of analytes to the substrate surface contributing to the background SERS signal. Herein, we propose a simple and effective nanofabrication procedure to produce large-scale 3D nanoporous Ag-based SERS substrates which exhibit excellent spatial reproducibility and EF of about 10 7 . Our substrates can detect polycyclic aromatic hydrocarbons dispersed in water without any functionalization of the surface, reaching a limit of detection (LOD) of ~20 nM for pyrene (PYR). As matter of fact, the nanoporous nature combines a high density of hotspots with the capacity to trap PAH molecules through not covalent bonding, facilitating their adhesion to the metallic surface. In perspective, our outcomes demonstrate the high potentiality of our approach for chemical sensing and environmental control applications.

We present a simple and effective nanofabrication procedure to produce large-scale 3D nanoporous Ag-based SERS substrates which exhibit excellent spatial reproducibility and EF of about 108. Our substrates can detect polycyclic aromatic hydrocarbons dispersed in water without any functionalization of the surface, reaching a LOD of ~ 20 nM.

Detection of analytes in aqueous solution with high specificity and sensitivity is of paramount importance in many fields of science, ranging from biomedicine, environmental control, and food quality assessment. Surface-enhanced Raman scattering (SERS) has proven to be a cutting-edge analytical technique for this purpose, by combining the high selectivity of Raman features with the high sensitivity deriving from the plasmonic amplification of Raman signals. Herein, we report a facile and quite effective approach to fabricate large-area Ag-based SERS substrates, exhibiting a porous, coral-like nanotexture. Due to their intrinsic large surface-area and high hot-spot density, the produced substrates appear quite promising for the detection of analytes at trace levels. The nanoporous substrates are produced by Solid-State Dewetting (SSD) of thin Ag-films. In particular, ~30 nm thickness Ag-films are first deposited on glass coverslips by magnetron sputtering. Then, marked roughening is induced by exposing the films to an Inductively Coupled Plasma (ICP) discharge, using synthetic air as feeding gas. The performances of our SERS substrates are characterized in terms of morphology and enhancement factor using CV as probe molecule.

The fabrication of plasmonic nanostructures with a reliable, low cost and easy approach has become a crucial and urgent challenge in many fields, including surface-enhanced Raman spectroscopy (SERS) based applications. In this frame, nanoporous metal films are quite attractive, due to their intrinsic large surface area and high density of metal nanogaps, acting as hot-spots for Raman signal enhancement. In this paper, we report a detailed study on the fabrication of nanoporous silver-based SERS substrates, obtained by the application of two successive treatments with an Inductively Coupled Plasma (ICP) system, using synthetic air and Ar as feeding gases. The obtained substrates exhibit a quite broad plasmonic response, covering the Vis-NIR range, and an enhancement factor reaching 6.5 [Formula: see text], estimated by using 4-mercaptobenzoic acid (4-MBA) as probe molecule at 532 nm. Moreover, the substrates exhibit a quite good spatial reproducibility on a centimeter scale, which assures a good signal stability for analytical measurements. Globally, the developed protocol is easy and cost effective, potentially usable also for mass production thanks to the remarkable inter-batches reproducibility. As such, it holds promise for its use in SERS-based sensing platforms for sensitive detection of targets molecules.

Hypothesis: While the phase behavior of aqueous surfactant solutions is usually described in term of the equilibrium microstructures of lyotropic liquid crystals, the transformations which take place when a phase turns into another one, either by changing the concentration or the temperature, are still to be elucidated. A simultaneous determination of concentration and microstructure is at order to elucidate the phase behavior under changing conditions, such as in a dissolution experiment. Experiments: Confocal Raman micro-spectroscopy and time-lapse polarized light microscopy are combined to study the phase transitions taking place in the dissolution of a common anionic surfactant (sodium laurylethersulfate) in water. Findings: By comparing Raman concentration profiles and polarized light images, it is found that the aqueous solution, with initial surfactant concentration of 72 wt%, undergoes a sequence of complex microstructural transformations including distortion of the initial lamellar phase, formation of an intermediate striated texture, which can be considered as a precursor of a cubic phase, and a heterogeneous hexagonal phase going through a transition region before turning into a micellar phase. The effects of the sodium counter-ion and of water confinement are also investigated by analyzing the OH-stretching bands. (C) 2019 Elsevier Inc. All rights reserved.

Tip-enhanced Raman spectroscopy is a powerful tool for the analysis of system interfaces, enabling access to chemical information with nanometric spatial resolution and sensitivity up to the single molecule level. Such features are due to the presence of proper metallic nanostructures at the TERS probe apex, which, via the excitation of a plasmonic field, confine light to a nanometric region. The nano-sized characteristic of such metallic structures intrinsically renders the fabrication of high performing and reproducible TERS probes still a challenge. In this paper, we present a facile, rapid and effective approach to prepare Ag-based TERS probes. The fabrication process proposed herein is based on spinodal dewetting of Ag-coated AFM-probes through a RF plasma treatment. The obtained probes appear covered with a coral-like silver nanotexture, endowed with an excellent plasmonic activity. Intriguingly, such a texture can be easily tuned by changing some process parameters, such as Ag film thickness and exposure time to the plasma. The as-prepared TERS probes show a high TERS enhancement, reaching 10(7), and allow a good spatial resolution, down to 10 nm. Finally, we suggest an easy and effective procedure to restore oxidized TERS tips following exposure to ambient air, which can be applied to all types of Ag-based TERS tips.

The phenomenon of glistenings, often appearing in intraocular lenses (IOLs) of patients after some time from the surgical operation, is potentially able to induce a poor quality of vision and, therefore, frustrate IOL implantation itself. In this paper, we combine optical microscopy with micro-Raman spectroscopy to get a deeper insight on the mechanism ruling, at microscopic scale, glistening formation. In particular, we have analyzed two types of IOLs, characterized by a different internal hydrophobicity but a similar polymer hydration coefficient. Raman imaging of single microvacuoles reveals that water creeps into the polymeric network, which traps water. Finally, applying the Principal Component Analysis (PCA) to Raman data, we provide information on the probable mechanism leading to water trapping in the two kinds of analyzed IOLs.

Tear fluid is a heterogeneous solution containing mainly proteins, lipids, mucins and electrolytes, which regulates the physiology of the human eye. The complex composition of tears can be altered in the presence of eye inflammations. The use of contact lenses is one of the most frequent causes of inflammatory responses of the eye, with the related discomfort often causing the wearer to give up using them. In this paper, we exploit the potentiality of Raman Spectroscopy to analyse the biochemical changes in tear fluid in a contact lens wearer. In particular, we analysed the tear fluid collected from a volunteer as a function of the wearing time for two types of monthly contact lenses (Hydrogel and Si-Hydrogel). Our experimental results show an alteration of the relative concentrations of proteins and lipids in both of the analysed cases. More importantly, our results highlight the diagnostic sensitivity of Raman analysis to select the proper contact lens type for each wearer and optimise the lens wearing conditions.

Sensitive detection of altered proteins expression in plasma membranes is of fundamental importance, for both diagnostic and prognostic purposes. Surface-Enhanced Raman Scattering (SERS) has proven to be a quite sensitive approach to detect proteins, even in very diluted samples. However, proteins detection in complex environment, such as the cellular membrane, is still a challenge. Herein, we demonstrate a SERS-based platform to reveal the overexpression of target proteins in cell membranes. As a proof of concept, we implemented ectopic expression of carbonic anhydrase IX (CA IX) and epidermal growth factor receptor (EGFR) in the plasma membrane of the SKOV3 tumor cell line. Our outcomes demonstrate that SERS signals from cells put in contact with a hyperuniform SERS substrate allow highlighting subtle differences in the biochemical composition of cell membranes, normally hidden in spontaneous Raman confocal microscopy. This opens new opportunities for a label-free membrane analysis and bio-sensing in a broader sense.