Evaluation of the SERS performances of Tabun and VX label-free detection in complex and multicomponent fluids.

Nerve agents represent a serious threat to security worldwide. Chemical terrorism has become an alarming danger since the technological progresses have simplified the production of nerve agents. Therefore, there is an immediate demand for a fast and precise detection of these compounds on-site and real-time. In this perspective, Surface-Enhanced Raman Scattering (SERS) has emerged as a well-suited alternative for on-field detection. SERS performances of unfunctionalized SERS substrates were evaluated in realistic samples. Two nerve agents, Tabun and VX, were diluted in two matrix models: a contact lens solution, and a caffeine-based eye serum. The performance two research-grade instruments and two portable devices were compared. Despite the use of a small sampling volume of complex matrices without any sample pre-treatment, we achieved Tabun detection in both media, with a practical limit of detection (LOD) in the range of 7-9 ppm in contact lens liquid, and of 10.2 ppm in eye serum. VX detection turned out to be more challenging and was achieved only in contact lens solution, with a practical LOD in the range of 0.6-5 ppm. These results demonstrate the feasibility of on-field detection of nerve agents with SERS, that could be implemented when there is suspicion of chemical threat.

Optics miniaturization strategy for demanding Raman spectroscopy applications.

Raman spectroscopy provides non-destructive, label-free quantitative studies of chemical compositions at the microscale as used on NASA's Perseverance rover on Mars. Such capabilities come at the cost of high requirements for instrumentation. Here we present a centimeter-scale miniaturization of a Raman spectrometer using cheap non-stabilized laser diodes, densely packed optics, and non-cooled small sensors. The performance is comparable with expensive bulky research-grade Raman systems. It has excellent sensitivity, low power consumption, perfect wavenumber, intensity calibration, and 7 cm-1 resolution within the 400-4000 cm-1 range using a built-in reference. High performance and versatility are demonstrated in use cases including quantification of methanol in beverages, in-vivo Raman measurements of human skin, fermentation monitoring, chemical Raman mapping at sub-micrometer resolution, quantitative SERS mapping of the anti-cancer drug methotrexate and in-vitro bacteria identification. We foresee that the miniaturization will allow realization of super-compact Raman spectrometers for integration in smartphones and medical devices, democratizing Raman technology.

Nitroaromatic explosives' detection and quantification using an attention-based transformer on surface-enhanced Raman spectroscopy maps.

Rapidly and accurately detecting and quantifying the concentrations of nitroaromatic explosives is critical for public health and security. Among existing approaches, explosives' detection with Surface-Enhanced Raman Spectroscopy (SERS) has received considerable attention due to its high sensitivity. Typically, a preprocessed single spectrum that is the average of the entire or a selected subset of a SERS map is used to train various machine learning models for detection and quantification. Designing an appropriate averaging and preprocessing procedure for SERS maps across different concentrations is time-consuming and computationally costly, and the averaging of spectra may lead to the loss of crucial spectral information. We propose an attention-based vision transformer neural network for nitroaromatic explosives' detection and quantification that takes raw SERS maps as the input without any preprocessing. We produce two novel SERS datasets, 2,4-dinitrophenols (DNP) and picric acid (PA), and one benchmark SERS dataset, 4-nitrobenzenethiol (4-NBT), which have repeated measurements down to concentrations of 1 nM to illustrate the detection limit. We experimentally show that our approach outperforms or is on par with the existing methods in terms of detection and concentration prediction accuracy. With the produced attention maps, we can further identify the regions with a higher signal-to-noise ratio in the SERS maps. Based on our findings, the molecule of interest detection and concentration prediction using raw SERS maps is a promising alternative to existing approaches.

Methotrexate Detection in Serum at Clinically Relevant Levels with Electrochemically Assisted SERS on a Benchtop, Custom Built Raman Spectrometer.

Therapeutic drug monitoring (TDM) is an essential clinical practice for optimizing drug dosing, thereby preventing adverse effects of drugs with a narrow therapeutic window, slow clearance, or high interperson pharmacokinetic variability. Monitoring methotrexate (MTX) during high-dose MTX (HD-MTX) therapy is necessary to avoid potentially fatal side effects caused by delayed elimination. Despite the efficacy of HD-MTX treatment, its clinical application in resource-limited settings is constrained due to the relatively high cost and time of analysis with conventional analysis methods. In this work, we developed (i) an electrochemically assisted surface-enhanced Raman spectroscopy (SERS) method for detecting MTX in human serum at a clinically relevant concentration range and (ii) a benchtop, Raman detection system with an integrated potentiostat, software, and data analysis unit that enables mapping of small areas of SERS substrates and quantitative SERS-based analysis. In the assay, by promoting electrostatic attraction between gold-coated nanopillar SERS substrates and MTX molecules in aqueous samples, a detection limit of 0.13 µM with a linear range of 0.43-2 µM was achieved in PBS. The implemented sample cleanup through gel filtration proved to be highly effective, resulting in a similar detection limit (0.55 µM) and linear range (1.81-5 µM) for both PBS and serum. The developed and optimized assay could also be used on the in-house built, Raman device. We showed that MTX detection can be carried out in less than 30 min with the Raman device, paving the way toward the TDM of MTX at the point-of-need and in resource-limited environments.

Quantification of Methotrexate in Human Serum Using Surface-Enhanced Raman Scattering-Toward Therapeutic Drug Monitoring.

Therapeutic drug monitoring (TDM) can improve clinical care when using drugs with pharmacokinetic variability and a narrow therapeutic window. Rapid, reliable, and easy-to-use detection methods are required in order to decrease the time of analysis and can also enable TDM in resource-limited settings or even at bedside. Monitoring methotrexate (MTX), an anticancer drug, is critical since it is needed to follow the drug clearance rate and decide how to administer the rescue drug, leucovorin (LV), in order to avoid toxicity and even death. We show that with the optimized nanopillar-assisted separation (NPAS) method using surface-enhanced Raman scattering, we were able to measure MTX in PBS and serum in the linear range of 5-150 µM and confirmed that MTX detection can be carried out even in the presence of LV. Additionally, when NPAS was combined with centrifugal filtration, a quantification limit of 2.1 µM for MTX in human serum sample was achieved. The developed detection method enables fast detection (10 min) and quantification of MTX from human serum (>90% accuracy). Furthermore, we show the potential of the developed method for TDM, when quantifying MTX from clinical samples, collected from patients who are undergoing high-dose MTX therapy.

Large plasmonic color metasurfaces fabricated by super resolution deep UV lithography.

In this paper, we demonstrate plasmonic color metasurfaces as large as ∼60 cm2 fabricated by deep UV projection lithography employing an innovative combination of resolution enhancement techniques. Briefly, in addition to the established off-axis dipole illumination, double- and cross-exposure resolution enhancement of lithography, we introduce a novel element, the inclusion of transparent assist features to the mask layout. With this approach, we demonstrate the fabrication of relief arrays having critical dimensions such as 159 nm nanopillars or 210 nm nanoholes with 300 nm pitches, which is near the theoretical resolution limit expressed by the Rayleigh criterion for the 248 nm lithography tool used in this work. The type of surface structure, i.e. nanopillar or nanohole, and their diameters can be tailored simply by changing the width of the assist features included in the mask layout. By coating the obtained nanopatterns with thin layers of either Au or Al, we observe color spectra originating from the phenomenon known as localized surface plasmon resonance (LSPR). We demonstrate the generation of color palettes representing a broad spectral range of colors, and we employ finite element modelling to corroborate the measured LSPR fingerprint spectra. Most importantly, the ∼60 cm2 nanostructure arrays can be written in only a few minutes, which is a tremendous improvement compared to the more established techniques employed for fabricating similar structures.

Controlled Drug Release from Biodegradable Polymer Matrix Loaded in Microcontainers Using Hot Punching.

Microcontainers are reservoir-based advanced drug delivery systems (DDS) that have proven to increase the bioavailibity of the small-molecule drugs, targeting of biomolecules, protection of vaccines and improved treatment of Pseudomonas aeruginosa. However, high-throughput loading of these micron-sized devices with drug has been challenging. Hot punching is a new technique that is a fast, simple and single-step process where the microdevices are themselves used as mold to punch biocompatible and biodegradable drug-polymer films, thereby loading the containers. Here, we investigate the effect of hot punching on the drug distribution as well as drug release from the loaded drug-polymer matrices. Zero-order sustained drug release is observed for the model drug Furosemide embedded in biodegradable polymer, Poly-ε-caprolactone, which is attributed to the unique spatial distribution of Furosemide during the loading process.

Experimental and First-Principles Spectroscopy of Cu2SrSnS4 and Cu2BaSnS4 Photoabsorbers.

Cu2BaSnS4 (CBTS) and Cu2SrSnS4 (CSTS) semiconductors have been recently proposed as potential wide band gap photovoltaic absorbers. Although several measurements indicate that they are less affected by band tailing than their parent compound Cu2ZnSnS4, their photovoltaic efficiencies are still low. To identify possible issues, we characterize CBTS and CSTS in parallel by a variety of spectroscopic methods complemented by first-principles calculations. Two main problems are identified in both materials. The first is the existence of deep defect transitions in low-temperature photoluminescence, pointing to a high density of bulk recombination centers. The second is their low electron affinity, which emphasizes the need for an alternative heterojunction partner and electron contact. We also find a tendency for downward band bending at the surface of both materials. In CBTS, this effect is sufficiently large to cause carrier-type inversion, which may enhance carrier separation and mitigate interface recombination. Optical absorption at room temperature is exciton-enhanced in both CBTS and CSTS. Deconvolution of excitonic effects yields band gaps that are about 100 meV higher than previous estimates based on Tauc plots. Although the two investigated materials are remarkably similar in an idealized, defect-free picture, the present work points to CBTS as a more promising absorber than CSTS for tandem photovoltaics.

Quantifying Optical Absorption of Single Plasmonic Nanoparticles and Nanoparticle Dimers Using Microstring Resonators.

The wide and ever-increasing applications of thermoplasmonics demand the need for sensitive and reliable tools to probe optical absorptions of individual nanoparticles. However, most of the currently available techniques focus only on measuring the surface temperature of nanostructures in a particular medium and are either invasive or suffer from low sensitivity, lengthy calibration, or the inability to probe single structures with nanogaps. Here, we present for the first time the use of micromechanical SiN string resonators for quantifying optical absorption cross sections of individual plasmonic nanostructures. Monomers and dimers of nanospheres, nanostars, shell-isolated nanoparticles, and nanocubes are probed. A reliable data treatment method is developed to obtain the absorption cross sections as a function of responsivity across a string. The presented method exhibits an excellent sensitivity of ∼89 Hz/K. This allows quantification of optical absorption cross sections of individual plasmonic structures even when their plasmon resonance wavelengths are far from the laser excitation wavelength. The experimentally obtained optical absorption cross sections agree well with the simulations. Influencing factors including polarization, surface morphology, and nanogap size are discussed. The developed method and the obtained optical absorption profiles facilitate future development and optimization of thermoplasmonic applications.

Selective surface-enhanced Raman scattering detection of Tabun, VX and Cyclosarin nerve agents using 4-pyridine amide oxime functionalized gold nanopillars.

We have earlier demonstrated sensitive detection of low the volatile nerve agents Tabun, Cyclosarin and VX by using handheld Raman instrumentation in conjunction with surface-enhanced Raman scattering (SERS) attained with gold and silver coated Si nanopillar substrates. In the present proof-of-concept study, the gold substrates chemically are functionalized to realize selectivity towards organophosphorus compounds (OPs) with high sensitivity. A potential capturer and reporter molecule, chemical nerve agent antidote, 4-pyridine amide oxime, is evaluated due to its high Raman cross section, high chemical affinity towards gold, and binding specificity to the target substances Tabun, VX and Cyclosarin via the oxime group. Upon selective and covalent binding, the SERS probe undergoes structural changes which are reflected in the spectral SERS responses, making it suitable for indirect monitoring of nerve agents in aqueous solution. With the probe attached to the hotspots of Au-coated Si nanopillars, the SERS signals distinctly discriminate between specific and non-specific analyte binding of Tabun, Cyclosarin and VX down to sub ppm levels. SERS spectrum of 4-PAO is measured after microliter drop coating of aqueous sample solution onto the functionalized substrates and subsequent water evaporation from surfaces. This binding assay is complemented by letting functionalized substrates being immersed into sample solutions 1 h before measuring. Binding specific SERS response decreases in following order: Tabun > VX > Cyclosarin. Overall, the concept looks promising, as expected the candidate probe 4-PAO introduces selectivity to the nanopillar gold substrates without loss of sensitivity.

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.

Quantitative SERS Assay on a Single Chip Enabled by Electrochemically Assisted Regeneration: A Method for Detection of Melamine in Milk.

Reusability of sensors is relevant when aiming to decrease variation between measurements, as well as cost and time of analysis. We present an electrochemically assisted surface-enhanced Raman spectroscopy (SERS) platform with the capability to reverse the analyte-surface interaction, without damaging the SERS substrate, allowing for efficient sensor reuse. The platform was used in combination with a sample pretreatment step, when detecting melamine from milk. We found that the electrochemically enhanced analyte-surface interaction results in significant improvement in detection sensitivity, with detection limits (0.01 ppm in PBS and 0.3 ppm in milk) below the maximum allowed levels in food samples. The reversibility of interaction enabled continuous measurement in aqueous solution and a complete quantitative assay on a single SERS substrate.

Nanopillar-Assisted SERS Chromatography.

Practical implementation of surfaced enhanced Raman spectroscopy (SERS) sensing is hindered by complexity of real-life samples, which often requires long and costly pretreatment and purification. Here, we present a novel nanopillar-assisted SERS chromatography (NPC-SERS) method for simultaneous quantitation of target molecules and analysis of complex, multicomponent fluids, e.g., human urine spiked with a model drug paracetamol (PAR). Gold-coated silicon nanopillar (AuNP) SERS substrates and a centrifugal microfluidic platform are tactfully combined, which allows (i) a precise and fully automated sample manipulation and (ii) spatial separation of different molecular species on the AuNP substrate. The NPC-SERS technique provides a novel approach for wetting the stationary phase (AuNP) using the "wicking effect", and thus minimizes dilution of analytes. Separation of PAR and the main human urine components (urea, uric acid, and creatinine) has been demonstrated. Quantitative detection of PAR with ultrawide linear dynamic range (0-500 ppm) is achieved by analyzing the spreading profiles of PAR on the AuNP surface. NPC-SERS transforms SERS into a sensing technique with general applicability, facilitating rapid and quantitative detection of analytes in complex biofluids, such as saliva, blood, and urine.

Label-Free Quantification of Anticancer Drug Imatinib in Human Plasma with Surface Enhanced Raman Spectroscopy.

Therapeutic drug monitoring (TDM) for anticancer drug imatinib has been suggested as the best way to improve the treatment response and minimize the risk of adverse reactions in chronic myelogenous leukemia (CML) and gastrointestinal stromal tumor (GIST) patients. TDM of oncology treatments with standard analytical methods, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS) is, however, complex and demanding. This paper proposes a new method for quantitation of imatinib in human plasma, based on surface enhanced raman spectroscopy (SERS) and multivariate calibration using partial least-squares regression (PLSR). The best PLSR model was obtained with three latent variables in the range from 123 to 5000 ng/mL of imatinib, providing a standard error of prediction (SEP) of 510 ng/mL. The method was validated in accordance with international guidelines, through the estimate of figures of merit, such as precision, accuracy, systematic error, analytical sensitivity, limits of detection, and quantitation. Moreover, the feasibility and clinical utility of this approach have also been verified using real plasma samples taken from deidentified patients. The results were in good agreement with a clinically validated LC-MS/MS method. The new SERS method presented in this preliminary work showed simplicity, short analysis time, good sensitivity, and could be considered a promising platform for TDM of imatinib treatment in a point-of-care setting.

Detecting forensic substances using commercially available SERS substrates and handheld Raman spectrometers.

Ultra-sensitive in-field measurements of most forensic substances still today remain a challenge for first responders and forensic investigators. Handheld Raman spectroscopy equipment is getting more and more routinely used in the field for evidence collection, however, restricted to measurements of pure or high concentration samples. Here, surface-enhanced Raman scattering (SERS) sensing of common forensic substances with commercially available SERS substrates and handheld spectrometers, have been investigated. 3D Finite Element Method (FEM) and Density Functional Theory (DFT) simulations were used to interpret the high SERS enhancement of the Ag nanopillar substrate and the detection of the substances, respectively. The forensic generality and high performance of the analytical method were demonstrated by explicit detection of close to unprecedented amounts, down to femtograms, of Cyclosarin, RDX, Amphetamine and Picric acid. Implications are ultra-sensitive in-field SERS detection of these substances with commercial equipment.

Large-Scale, Lithography-Free Production of Transparent Nanostructured Surface for Dual-Functional Electrochemical and SERS Sensing.

In this work, we present a dual-functional sensor that can perform surface-enhanced Raman spectroscopy (SERS) based identification and electrochemical (EC) quantification of analytes in liquid samples. A lithography-free reactive ion etching process was utilized to obtain nanostructures of high aspect ratios distributed homogeneously on a 4 in. fused silica wafer. The sensor was made up of three-electrode array, obtained by subsequent e-beam evaporation of Au on nanostructures in selected areas through a shadow mask. The SERS performance was evaluated through surface-averaged enhancement factor (EF), which was ∼6.2 × 105, and spatial uniformity of EF, which was ∼13% in terms of relative standard deviation. Excellent electrochemical performance and reproducibility were revealed by recording cyclic voltammograms. On nanostructured electrodes, paracetamol (PAR) showed an improved quasi-reversible behavior with decrease in peak potential separation (ΔEp ∼ 90 mV) and higher peak currents (Ipa/Ipc ∼ 1), compared to planar electrodes (ΔEp ∼ 560 mV). The oxidation potential of PAR was also lowered by ∼80 mV on nanostructured electrodes. To illustrate dual-functional sensing, quantitative evaluation of PAR ranging from 30 µM to 3 mM was realized through EC detection, and the presence of PAR was verified by its SERS fingerprint.

Nanopillar Filters for Surface-Enhanced Raman Spectroscopy.

We present a simple, robust, and automated molecule extraction technique based on a centrifugal microfluidic platform. Fast and facile extraction of a food adulterant (melamine) from a complex sample medium (milk) on a SERS substrate is demonstrated. The unique characteristic of the detection method is the obtained "filter paper/chromatographic" effect which combines centrifugal force and wetting properties of the SERS substrate. The work addresses issues related to SERS-based detection of analytes in complex media, which is important for realizing next generation SERS platforms applicable for a broad variety of real-life applications.

Hand-Held Femtogram Detection of Hazardous Picric Acid with Hydrophobic Ag Nanopillar SERS Substrates and Mechanism of Elasto-Capillarity.

Picric acid (PA) is a severe environmental and security risk due to its unstable, toxic, and explosive properties. It is also challenging to detect in trace amounts and in situ because of its highly acidic and anionic character. Here, we assess sensing of PA under nonlaboratory conditions using surface-enhanced Raman scattering (SERS) silver nanopillar substrates and hand-held Raman spectroscopy equipment. The advancing elasto-capillarity effects are explained by molecular dynamics simulations. We obtain a SERS PA detection limit on the order of 20 ppt, corresponding attomole amounts, which together with the simple analysis methodology demonstrates that the presented approach is highly competitive for ultrasensitive analysis in the field.

Surface Enhanced Raman Scattering for Quantification of p-Coumaric Acid Produced by Escherichia coli.

The number of newly developed genetic variants of microbial cell factories for production of biochemicals has been rapidly growing in recent years, leading to an increased need for new screening techniques. We developed a method based on surface-enhanced Raman scattering (SERS) coupled with liquid-liquid extraction (LLE) for quantification of p-coumaric acid (pHCA) in the supernatant of genetically engineered Escherichia coli (E. coli) cultures. pHCA was measured in a dynamic range from 1 µM up to 50 µM on highly uniform SERS substrates based on leaning gold-capped nanopillars, which showed an in-wafer signal variation of only 11.7%. LLE using dichloromethane as organic phase was combined with the detection in order to increase selectivity and sensitivity by decreasing the effect of interfering compounds from the analytes of interest. The difference in pHCA production yield between three genetically engineered E. coli strains was successfully evaluated using SERS and confirmed with high-performance liquid chromatography. As this novel approach has potential to be automated and parallelized, it can be considered for high-throughput screening in metabolic engineering.

SERS detection of the biomarker hydrogen cyanide from Pseudomonas aeruginosa cultures isolated from cystic fibrosis patients.

Pseudomonas aeruginosa is the primary cause of chronic airway infections in cystic fibrosis (CF) patients. Persistent infections are seen from the first P. aeruginosa culture in about 75% of young CF patients, and it is important to discover new ways to detect P. aeruginosa at an earlier stage. The P. aeruginosa biomarker hydrogen cyanide (HCN) contains a triple bond, which is utilized in this study because of the resulting characteristic C≡N peak at 2135 cm-1 in a Raman spectrum. The Raman signal was enhanced by surface-enhanced Raman spectroscopy (SERS) on a Au-coated SERS substrate. After long-term infection, a mutation in the patho-adaptive lasR gene can alter the expression of HCN, which is why it is sometimes not possible to detect HCN in the breath of chronically infected patients. Four P. aeruginosa reference strains and 12 clinical P. aeruginosa strains isolated from CF children were evaluated, and HCN was clearly detected from overnight cultures of all wild type-like isolates and half of the later isolates from the same patients. The clinical impact could be that P. aeruginosa infections could be detected at an earlier stage, because daily breath sampling with an immediate output could be possible with a point-of-care SERS device.