Chiroplasmonic DNA Scaffolded "Fusilli" Structures.

DNA is an ideal template for the design of nanoarchitectures with molecular-like features. Here, we present an optimized assembly strategy for the concatenation of DNA quasi-rings into long scaffolds. Ionic strength, which played a major role during self-assembly, produced the expected high quality only at 15 mM MgCl2. Atomic force microscopy (AFM) characterization showed several micrometer long tubular structures that were used as templates for the positioning of plasmonic nanoparticles (NPs) along a three-dimensional helical path using DNA tethers. As imaged by high-resolution scanning transmission electron microscopy (HR-STEM) and modeled by theoretical calculations, the NPs distributed into a "fusilli" fashion (i.e., a helical pasta shape), displaying chiroptical activity as revealed by a bisignated CD absorption, centered at the plasmon resonance wavelength. The present structures contribute to enrich the ever-developing arena of chiroplasmonic DNA-based nanomaterials and demonstrate that large assemblies are attainable for their future application to develop metamaterials.

Surface Enhanced Raman Spectroscopy for Quantitative Analysis: Results of a Large-Scale European Multi-Instrument Interlaboratory Study.

Surface-enhanced Raman scattering (SERS) is a powerful and sensitive technique for the detection of fingerprint signals of molecules and for the investigation of a series of surface chemical reactions. Many studies introduced quantitative applications of SERS in various fields, and several SERS methods have been implemented for each specific application, ranging in performance characteristics, analytes used, instruments, and analytical matrices. In general, very few methods have been validated according to international guidelines. As a consequence, the application of SERS in highly regulated environments is still considered risky, and the perception of a poorly reproducible and insufficiently robust analytical technique has persistently retarded its routine implementation. Collaborative trials are a type of interlaboratory study (ILS) frequently performed to ascertain the quality of a single analytical method. The idea of an ILS of quantification with SERS arose within the framework of Working Group 1 (WG1) of the EU COST Action BM1401 Raman4Clinics in an effort to overcome the problematic perception of quantitative SERS methods. Here, we report the first interlaboratory SERS study ever conducted, involving 15 laboratories and 44 researchers. In this study, we tried to define a methodology to assess the reproducibility and trueness of a quantitative SERS method and to compare different methods. In our opinion, this is a first important step toward a "standardization" process of SERS protocols, not proposed by a single laboratory but by a larger community.

Predictions on the SERS enhancement factor of gold nanosphere aggregate samples.

Colloidal gold nanostructures are nowadays widely involved in sensor applications. One of the most interesting techniques that takes advantage of them is certainly the Surface Enhanced Raman Scattering (SERS) effect, even if it is often considered a tricky technique due to structural constraints required by the nanostructured substrates to obtain high enhancement factors (EFs), i.e. the presence of hot spots. Because of the easy preparation and high number of hot spots, aggregated gold nanospheres seem to be the most efficient through the SERS colloids, but their characteristic high disorder makes them unpredictable and difficult to compare between different batches. For this reason, less SERS effective, but more regular and organized substrates are usually preferred. In this study, a method based on Boundary Element Method (BEM) simulation is used to accurately predict the colloidal SERS EFs of gold nanoparticle (AuNP) aggregates, starting from their experimental extinction spectra. Surprisingly, it was found that larger aggregates do not exhibit stronger hot spots, but rather higher amounts of them, influencing the overall predicted EFs, which well reflect the results obtained experimentally.

Detection of low-quantity anticancer drugs by surface-enhanced Raman scattering.

Ultrasensitive detection of low-quantity drugs is important for personalized therapeutic approaches in several diseases and, in particular, for cancer treatment. In this field, surface-enhanced Raman scattering (SERS) can be very useful for its ability to precisely identify analytes from their unique vibrational spectra, with very high sensitivity. Here, we report a study about SERS detection of sunitinib, paclitaxel and irinotecan, i.e. three commonly used antineoplastic drugs, and of SN-38, i.e. the metabolite of irinotecan, dissolved in methanol solutions. By using commercial Klarite substrates, we found that sunitinib, irinotecan and SN-38 have detection limits of 20-70 ng, which is below the threshold for applications in cancer therapy. Conversely, the SERS signal was not appreciable with paclitaxel, and this is explained by the absence of optical resonances in the visible range. Overall, our results show that ultrasensitive SERS detection of sunitinib, irinotecan and SN-38 is feasible, encouraging further development of this technology also for other drugs with similar molecular structure especially for those analytes with absorption bands in the visible range.

Magneto-plasmonic Au-Fe alloy nanoparticles designed for multimodal SERS-MRI-CT imaging.

Diagnostic approaches based on multimodal imaging are needed for accurate selection of the therapeutic regimens in several diseases, although the dose of administered contrast drugs must be reduced to minimize side effects. Therefore, large efforts are deployed in the development of multimodal contrast agents (MCAs) that permit the complementary visualization of the same diseased area with different sensitivity and different spatial resolution by applying multiple diagnostic techniques. Ideally, MCAs should also allow imaging of diseased tissues with high spatial resolution during surgical interventions. Here a new system based on multifunctional Au-Fe alloy nanoparticles designed to satisfy the main requirements of an ideal MCA is reported and their biocompatibility and imaging capability are described. The MCAs show easy and versatile surface conjugation with thiolated molecules, magnetic resonance imaging (MRI) and computed X-ray tomography (CT) signals for anatomical and physiological information (i.e., diagnostic and prognostic imaging), large Raman signals amplified by surface enhanced Raman scattering (SERS) for high sensitivity and high resolution intrasurgical imaging, biocompatibility, exploitability for in vivo use and capability of selective accumulation in tumors by enhanced permeability and retention effect. Taken together, these results show that Au-Fe nanoalloys are excellent candidates as multimodal MRI-CT-SERS imaging agents.

SERS nanostructures with engineered active peptides against an immune checkpoint protein.

The immune checkpoint programmed death ligand 1 (PD-L1) protein is expressed by tumor cells and it suppresses the killer activity of CD8+ T-lymphocyte cells binding to the programmed death 1 (PD-1) protein of these immune cells. Binding to either PD-L1 or PD1 is used for avoiding the inactivation of CD8+ T-lymphocyte cells. We report, for the first time, Au plasmonic nanostructures with surface-enhanced Raman scattering (SERS) properties (SERS nanostructures) and functionalized with an engineered peptide (CLP002: Trp-His-Arg-Ser-Tyr-Tyr-Thr-Trp-Asn-Leu-Asn-Thr), which targets PD-L1. Molecular dynamics calculations are used to describe the interaction of the targeting peptide with PD-L1 in the region where the interaction with PD-1 occurs, showing also the poor targeting activity of a peptide with the same amino acids, but a scrambled sequence. The results are confirmed experimentally since a very good targeting activity is observed against the MDA-MB-231 breast adenocarcinoma cancer cell line, which overexpresses PD-L1. A good activity is observed, in particular, for SERS nanostructures where the CLP002-engineered peptide is linked to the nanostructure surface with a short charged amino acid sequence and a long PEG chain. The results show that the functionalized SERS nanostructures show very good targeting of the immune checkpoint PD-L1.

LDI-MS assisted by chemical-free gold nanoparticles: enhanced sensitivity and reduced background in the low-mass region.

Gold nanoparticles (AuNPs) assisted laser desorption ionization mass spectrometry (LDI-MS) emerged as an effective technique for the detection of analytes with high sensitivity. The surface chemistry and the size of AuNPs are the crucial parameters for lowering the detection limits and increasing the selectivity of LDI-MS. Here we show that chemical-free size selected AuNPs, obtained by laser ablation synthesis in solution (LASiS), have very low background in the low mass region (<500 Da), contrary to citrate stabilized AuNPs (citrate-AuNPs) and dihydroxyacetophenone (DHAP). This allowed better performances for the picomole detection of low mass analytes like arginine, fructose, atrazine, anthracene and paclitaxel. The results suggest that chemical-free LASiS-AuNPs can be an excellent matrix for nanoparticle-assisted LDI-MS.

Alternative SERRS probes for the immunochemical localization of ovalbumin in paintings: an advanced mapping detection approach.

In the field of analytical chemistry, many scientific efforts have been devoted to develop experimental procedures for the characterization of organic substances present in heterogeneous artwork samples, due to their challenging identification. In particular, performances of immunochemical techniques have been recently investigated, optimizing ad hoc systems for the identification of proteins. Among all the different immunochemical approaches, the use of metal nanoparticles - for surface enhanced Raman scattering (SERS) detection - remains one of the most powerful methods that has still not been explored enough for the analysis of artistic artefacts. For this reason, the present research work was aimed at proposing a new optimized and highly efficient indirect immunoassay for the detection of ovalbumin. In particular, the study proposed a new SERRS probe composed of gold nanoparticles (AuNPs) functionalised with Nile Blue A and produced with an excellent green and cheap alternative approach to the traditional chemical nanoparticles synthesis: the laser ablation synthesis in solution (LASiS). This procedure allows us to obtain stable nanoparticles which can be easily functionalized without any ligand exchange reaction or extensive purification procedures. Moreover, the present research work also focused on the development of a comprehensive analytical approach, based on the combination of potentialities of immunochemical methods and Raman analysis, for the simultaneous identification of the target protein and the different organic and inorganic substances present in the paint matrix. An advanced mapping detection system was proposed to achieve the exact spatial location of all the components through the creation of false colour chemical maps.

High performance multi-purpose nanostructured thin films by inkjet printing: Au micro-electrodes and SERS substrates.

Nanostructured thin metal films are exploited in a wide range of applications, spanning from electrical to optical transducers and sensors. Inkjet printing has become a compliant technique for sustainable, solution-processed, and cost-effective thin films fabrication. Inspired by the principles of green chemistry, here we show two novel formulations of Au nanoparticle-based inks for manufacturing nanostructured and conductive thin films by using inkjet printing. This approach showed the feasibility to minimize the use of two limiting factors, namely stabilizers and sintering. The extensive morphological and structural characterization provides pieces of evidence about how the nanotextures lead to high electrical and optical performances. Our conductive films (sheet resistance equal to 10.8 ± 4.1 Ω per square) are a few hundred nanometres thick and feature remarkable optical properties in terms of SERS activity with enhancement factors as high as 107 averaged on the mm2 scale. Our proof-of-concept succeeded in simultaneously combining electrochemistry and SERS by means of real-time tracking of the specific signal of mercaptobenzoic acid cast on our nanostructured electrode.

Identification and quantification of tire wear particles by employing different cross-validation techniques: FTIR-ATR Micro-FTIR, Pyr-GC/MS, and SEM.

Tire wear particles (TWPs) are one of the environment's most important emission sources of microplastics. In this work, chemical identification of these particles was carried out in highway stormwater runoff through cross-validation techniques for the first time. Optimization of a pre-treatment method (i.e., extraction and purification) was provided to extract TWPs, avoiding their degradation and denaturation, to prevent getting low recognizable identification and consequently underestimates in the quantification. Specific markers were used for TWPs identification comparing real stormwater samples and reference materials via FTIR-ATR, Micro-FTIR, and Pyrolysis-gas-chromatography-mass spectrometry (Pyr-GC/MS). Quantification of TWPs was carried out via Micro-FTIR (microscopic counting); the abundance ranged from 220,371 ± 651 TWPs/L to 358,915 ± 831 TWPs/L, while the higher mass was 39,6 ± 9 mg TWPs/L and the lowest 31,0 ± 8 mg TWPs/L. Most of the TWPs analyzed were less than 100 µm in size. The sizes were also confirmed using a scanning electron microscope (SEM), including the presence of potential nano TWPs in the samples. Elemental analysis via SEM supported that a complex mixture of heterogeneous composition characterizes these particles by agglomerating organic and inorganic particles that could derive from brake and road wear, road pavement, road dust, asphalts, and construction road work. Due to the analytical lack of knowledge about TWPs chemical identification and quantification in scientific literature, this study significantly contributes to providing a novel pre-treatment and analytical methodology for these emerging contaminants in highway stormwater runoff. The results of this study highlight the uttermost necessity to employ cross-validation techniques, i.e., FTIR-ATR, Micro-FTIR, Pyr-GC/MS, and SEM for the TWPs identification and quantification in the real environmental samples.

Mechanical and Tribological Properties of Ta-N and Ta-Al-N Coatings Deposited by Reactive High Power Impulse Magnetron Sputtering.

In this article, the depositions and functional characterizations of Ta-N and Ta-Al-N coatings for protection purposes, grown by reactive high-power impulse magnetron sputtering onto silicon substrates, are described. Nitride films were grown while changing the substrate polarization voltage (i.e., the applied bias voltage) during the process. Moreover, the effects of adding Al to form a ternary system and the resulting variation of the coatings' mechanical and tribological properties have been widely investigated by nanoindentation, scratch, and wear tests. Micro-Raman characterization has been applied to the wear tracks to explore the comprehensive tribo-environment and wear mechanism. Interestingly, Ta-Al-N films, despite significantly improved mechanical properties, show a premature failure with respect to Ta-N coatings. The wear mechanisms of Ta-N and Ta-Al-N systems were revealed to be very different. Indeed, Ta-Al-N films suffer higher oxidation phenomena during wear, with the formation of an oxidized surface tribofilm and a reduced wear resistance, while Ta-N coatings undergo plastic deformation at the wear surface, with a slightly adhesive effect.

Enhancement of Magnetic Surface-Enhanced Raman Scattering Detection by Tailoring Fe3O4@Au Nanorod Shell Thickness and Its Application in the On-site Detection of Antibiotics in Water.

Surface-enhanced Raman scattering (SERS) has become a promising method for the detection of contaminants or biomolecules in aqueous media. The low interference of water, the unique spectral fingerprint, and the development of portable and handheld equipment for in situ measurements underpin its predominance among other spectroscopic techniques. Among the SERS nanoparticle substrates, those composed of plasmonic and magnetic components are prominent examples of versatility and efficiency. These substrates harness the ability to capture the target analyte, concentrate it, and generate unique hotspots for superior enhancement. Here, we have evaluated the use of gold-coated magnetite nanorods as a novel multifunctional magnetic-plasmonic SERS substrate. The nanostructures were synthesized starting from core-satellite structures. A series of variants with different degrees of Au coatings were then prepared by seed-mediated growth of gold, from core-satellite structures to core-shell with partial and complete shells. All of them were tested, using a portable Raman instrument, with the model molecule 4-mercaptobenzoic acid in colloidal suspension and after magnetic separation. Experimental results were compared with the boundary element method to establish the mechanism of Raman enhancement. The results show a quick magnetic separation of the nanoparticles and excellent Raman enhancement for all the nanoparticles both in dispersion and magnetically concentrated with limits of detection up to the nM range (∼50 nM) and a quantitative calibration curve. The nanostructures were then tested for the sensing of the antibiotic ciprofloxacin, highly relevant in preventing antibiotic contaminants in water reservoirs and drug monitoring, showing that ciprofloxacin can be detected using a portable Raman instrument at a concentration as low as 100 nM in a few minutes, which makes it highly relevant in practical point-of-care devices and in situ use.

3D Printed Microfluidic Device for Magnetic Trapping and SERS Quantitative Evaluation of Environmental and Biomedical Analytes.

Surface-enhanced Raman scattering (SERS) is an ideal technique for environmental and biomedical sensor devices due to not only the highly informative vibrational features but also to its ultrasensitive nature and possibilities toward quantitative assays. Moreover, in these areas, SERS is especially useful as water hinders most of the spectroscopic techniques such as those based on IR absorption. Despite its promising possibilities, most SERS substrates and technological frameworks for SERS detection are still restricted to research laboratories, mainly due to a lack of robust technologies and standardized protocols. We present herein the implementation of Janus magnetic/plasmonic Fe3O4/Au nanostars (JMNSs) as SERS colloidal substrates for the quantitative determination of several analytes. This multifunctional substrate enables the application of an external magnetic field for JMNSs retention at a specific position within a microfluidic channel, leading to additional amplification of the SERS signals. A microfluidic device was devised and 3D printed as a demonstration of cheap and fast production, with the potential for large-scale implementation. As low as 100 µL of sample was sufficient to obtain results in 30 min, and the chip could be reused for several cycles. To show the potential and versatility of the sensing system, JMNSs were exploited with the microfluidic device for the detection of several relevant analytes showing increasing analytical difficulty, including the comparative detection of p-mercaptobenzoic acid and crystal violet and the quantitative detection of the herbicide flumioxazin and the anticancer drug erlotinib in plasma, where calibration curves within diagnostic concentration intervals were obtained.

Wavy graphene sheets from electrochemical sewing of corannulene.

The presence of non-hexagonal rings in the honeycomb carbon arrangement of graphene produces rippled graphene layers with valuable chemical and physical properties. In principle, a bottom-up approach to introducing distortion from planarity of a graphene sheet can be achieved by careful insertion of curved polyaromatic hydrocarbons during the growth of the lattice. Corannulene, the archetype of such non-planar polyaromatic hydrocarbons, can act as an ideal wrinkling motif in 2D carbon nanostructures. Herein we report an electrochemical bottom-up method to obtain egg-box shaped nanographene structures through a polycondensation of corannulene that produces a new conducting layered material. Characterization of this new polymeric material by electrochemistry, spectroscopy, electron microscopy (SEM and TEM), scanning probe microscopy, and laser desorption-ionization time of flight mass spectrometry provides strong evidence that the anodic polymerization of corannulene, combined with electrochemically induced oxidative cyclodehydrogenations (Scholl reactions), leads to polycorannulene with a wavy graphene-like structure.

SERRS multiplexing with multivalent nanostructures for the identification and enumeration of epithelial and mesenchymal cells.

Liquid biopsy represents a new frontier of cancer diagnosis and prognosis, which allows the isolation of tumor cells released in the blood stream. The extremely low abundance of these cells needs appropriate methodologies for their identification and enumeration. Herein we present a new protocol based on surface enhanced resonance Raman scattering (SERRS) gold multivalent nanostructures to identify and enumerate tumor cells with epithelial and mesenchimal markers. The validation of the protocol is obtained with spiked samples of peripheral blood mononuclear cells (PBMC). Gold nanostructures are functionalized with SERRS labels and with antibodies to link the tumor cells. Three types of such nanosystems were simultaneously used and the protocol allows obtaining the identification of all individual tumor cells with the help of a Random Forest ensemble learning method.

Graphite-Based Geothermometry on Almahata Sitta Ureilitic Meteorites.

The thermal history of carbon phases, including graphite and diamond, in the ureilite meteorites has implications for the formation, igneous evolution, and impact disruption of their parent body early in the history of the Solar System. Geothermometry data were obtained by micro-Raman spectroscopy on graphite in Almahata Sitta (AhS) ureilites AhS 72, AhS 209b and AhS A135A from the University of Khartoum collection. In these samples, graphite shows G-band peak centers between 1578 and 1585 cm-1 and the full width at half maximum values correspond to a crystallization temperature of 1266 °C for graphite for AhS 209b, 1242 °C for AhS 72, and 1332 °C for AhS A135A. Recent work on AhS 72 and AhS 209b has shown graphite associated with nanodiamonds and argued that this assemblage formed due to an impact-event. Our samples show disordered graphite with a crystalline domain size ranging between about 70 and 140 nm. The nanometric grain-size of the recrystallized graphite indicates that it records a shock event and thus argues that the temperatures we obtained are related to such an event, rather than the primary igneous processing of the ureilite parent body.

Nanoparticles Engineering by Pulsed Laser Ablation in Liquids: Concepts and Applications.

Laser synthesis emerges as a suitable technique to produce ligand-free nanoparticles, alloys and functionalized nanomaterials for catalysis, imaging, biomedicine, energy and environmental applications. In the last decade, laser ablation and nanoparticle generation in liquids has proven to be a unique and efficient technique to generate, excite, fragment and conjugate a large variety of nanostructures in a scalable and clean way. In this work, we give an overview on the fundamentals of pulsed laser synthesis of nanocolloids and new information about its scalability towards selected applications. Biomedicine, catalysis and sensing are the application areas mainly discussed in this review, highlighting advantages of laser-synthesized nanoparticles for these types of applications and, once partially resolved, the limitations to the technique for large-scale applications.

A surface enhanced Raman scattering based colloid nanosensor for developing therapeutic drug monitoring.

Competitive reactions, on the surface of plasmonic nanostructures, allow exploiting SERS signals for quantitative Therapeutic Drug Monitoring. As an example, the concentration of Erlotinib, an anti-EGFR small molecule, used for the treatment of non-small cell lung and pancreatic cancer, is determined. The numerous side effects and the variability of patient responses make Erlotinib a good candidate for monitoring. The new SERS based sensor can estimate Erlotinib down to nanomolar concentration and is based on the chemical reaction of the drug and of a competitor SERS reporter on the surface of gold nanostructures. Colloid solutions of naked gold nanoparticles obtained by laser ablation in solution were used for obtaining nanostructures with very efficient hot spots for SERS and with a clean surface for chemistry. Detection of the drug in the nanomolar concentration range is shown to be possible also in spiked plasma samples.

Thiolated Graphene Oxide Nanoribbons as Templates for Anchoring Gold Nanoparticles: Two-Dimensional Nanostructures for SERS.

Graphene oxide nanoribbons (GONRs), obtained from the oxidative unzipping of carbon nanotubes, have been investigated as building blocks towards reaching active platforms in surface-enhanced Raman scattering (SERS). The complete development of carbon nanomaterials is strongly related to the exploitation of their chemical versatility, so this work is focused on the positive effect that a specific chemical functionalization provides to the SERS effect when gold nanoparticles are used. The covalent derivatization of GONRs with terminal thiol groups boosts their interaction with different types of gold nanoparticles (namely, 'naked' or citrate-stabilized), and the resulting two-dimensional aggregates show an intense enhancement of the Raman scattering from the carbon nanostructures because of their two-dimensional extended aggregation pattern. The SERS effect has been corroborated by theoretical calculations and a conceptual proof of SERS-based sensing.

PreS1 peptide-functionalized gold nanostructures with SERRS tags for efficient liver cancer cell targeting.

Early detection is the most effective mean of improving prognosis for many fatal diseases such as cancer. In this context, the Surface Enhanced Resonance Raman Scattering (SERRS) technique is being proposed as alternative to fluorescent methods in detection of biomarkers, because SERRS nanostructures are bright as fluorescent tags but more stable and clearly detectable using the narrow Raman "fingerprints" of a suitable reporter. Here we show that biocompatible SERRS active gold nanostructures, functionalized with an engineered PreS1 peptide (AuNP@PEG-PreS1), detect the presence of the SerpinB3 antigen overexpressed on liver tumor cells, a biomarker of the onset of liver cell carcinomatous transformation. A proper engineering of the targeting unit, linked to the nanostructure by a polymer chain, affords a sensitivity and specificity larger than 80%, at subnanomolar concentrations. Taking into account the high sensitivity of SERRS and that SB3 overexpression is an early event in liver cell carcinomatous transformation, AuNP@PEG-PreS1 nanostructures could be used in routine diagnostic activities, to improve the accuracy of HCC detection in particular in patients with chronic liver diseases.