Real-Time Monitoring of Molecules in Aqueous Solution via a Surface-Functionalized Ag-Anodic Aluminum Oxide Surface-Enhanced Raman Scattering Platform.

Real-time monitoring of molecular species in aqueous solutions is crucial for diverse scientific applications, from biomedical diagnostics to environmental analysis. In this study, we investigate the selective detection and discrimination of specific molecules in aqueous solution samples using a Ag-coated anodized aluminum oxide (Ag-AAO) surface functionalized with thiol molecules. Our investigation harnesses the power of surface-enhanced Raman scattering (SERS) synergized with principal component analysis (PCA) to elucidate the distinctive signatures of aqueous dopamine and l-tyrosine molecules. By scrutinizing the Raman spectra of surface-treated molecules, we unveil nuanced variations driven by the unique functional groups of the thiol molecules and their dynamic interactions with the target molecules in solution. Notably, we observe different alterations in the SERS spectra of Ag-AAO surface-functionalized boronic acid molecules for detection of dopamine and l-tyrosine, even at a concentration as low as 10-8 M. Moreover, the spectral PCA elucidates the discrimination of dopamine and l-tyrosine within the aqueous environment attributed to the different molecular interactions near SERS-active hotspots. Our findings facilitate real-time monitoring of minute analytes with exceptional molecular selectivity, ushering in an era of precise chemical analysis in aqueous solutions.

Label-Free Exosome Analysis by Surface-Enhanced Raman Scattering Spectroscopy with Laser-Ablated Silver Nanoparticle Substrate.

Early diagnostics of breast cancer is crucial to reduce the risk of cancer metastasis and late relapse. Exosome, which contains distinct information of its origin, can be the target object as a liquid biopsy. However, its low sensitivity and inadequate diagnostic tools interfere with the point-of-care testing (POCT) of the exosome. Recently, Surface-enhanced Raman Scattering (SERS) spectroscopy, which amplifies the Raman scattering, has been proved as a promising tool for exosome detection. However, the fabrication process of SERS probe or substrate is still inefficient and far from large-scale production. This study proposes rapid and label-free detection of breast cancer-derived exosomes by statistical analysis of SERS spectra using silver-nanoparticle-based SERS substrate fabricated by selective laser ablation and melting (SLAM). Employing silver nanowires and optimizing laser process parameters enable rapid and low-energy fabrication of SERS substrate. The functionalities including sensitivity, reproducibility, stability, and renewability are evaluated using rhodamine 6G as a probe molecule. Then, the feasibility of POCT is examined by the statistical analysis of SERS spectra of exosomes from malignant breast cancer cells and non-tumorigenic breast epithelial cells. The presented framework is anticipated to be utilized in other biomedical applications, facilitating cost-effective and large-scale production performance.

Au/Ag@ZIF-8 nanocomposite as solid phase extraction adsorbent and SERS substrate for tacrolimus label-free therapeutic drug monitoring in human serum.

Surface Enhanced Raman Scattering (SERS) has been extensively utilized in therapeutic drug monitoring (TDM) due to its rapid detection speed, high sensitivity and straightforward sample pretreatment. In this study, Au/AgNPs were obtained through the reduction of AgNO3 on the surface of AuNPs. Subsequently, Au/AgNPs were embedded into the tetrahedral lattice of ZIF-8 MOFs, resulting in the formation of Au/Ag@ZIF-8 nanocomposites. The Au/Ag@ZIF-8 nanocomposites exhibit a robust electromagnetic enhancement of Au/Ag bimetallic nanoparticles and a considerable adsorption capacity of ZIF-8 MOFs. This enables the pre-enrichment of target molecules in the vicinity of the electromagnetic field of the Au/AgNPs, thereby enhancing the sensitivity of SERS detection. The SERS substrate also exhibits high stability and reproducibility, as well as molecular sieving effects, due to the fact that Au/AgNPs are embedded into the tetrahedral lattice of ZIF-8. A TDM method for tacrolimus (FK506) in human serum was developed by using Au/Ag@ZIF-8 nanocomposites as solid phase extraction (SPE) adsorbent and SERS substrates. The results showed that under the optimized conditions, tacrolimus exhibited satisfactory linearity within the concentration range of 10-5-10-11 mol L-1, with a correlation coefficient (R2) of 0.9944, and the limit of detection (LOD) was as low as 6.4 pg mL-1. The recoveries were observed to range between 92 % and 105 %, with an RSD of below 8 %. The method is highly sensitive, exhibiting a sensitivity that is 3-6 orders of magnitude higher than that of existing analytical techniques. It has the potential to be applied in a clinical setting to biological samples.

Rapid determination of phenolic composition in chamomile (Matricaria recutita L.) using surface-enhanced Raman spectroscopy.

Flavonoids and hydroxycinnamic acids are the main responsible of the antioxidant activity of chamomile (Matricaria recutita L.). Traditional methods for the analysis of the phenolic content in vegetables often suffer from limitations such as being expensive, time-consuming, and complex. In this study, we propose, for the first time, the use of surface-enhanced Raman spectroscopy (SERS) for the rapid determination of the main components of the polyphenolic fraction in chamomile. Results demonstrate that SERS can serve as an alternative or complementary technique to main analytical strategies for qualitative and quantitative determination of polyphenol compounds in plant extracts. The method can be proposed for quasi real-time analysis of herbal teas and infusions, facilitating rapid screening of their main antioxidant components.

Ultrasensitive detection of contaminants in milk using a novel NMS-Ag modified water-resistant paper substrate.

Rapid and precise detection of harmful substances in food products is essential for ensuring public health and safety. This study introduces a novel surface-enhanced Raman spectroscopy (SERS) substrate, composed of a molybdenum disulfide­silver nanocomposite, applied to flexible, water-resistant filter paper for detecting melamine and bisphenol A (BPA) in milk. Optimized molybdenum disulfide (NMS) nanoflowers (NFs) were synthesized through hydrothermal methods and high-temperature annealing, then modified with silver (Ag) nanoparticles to form the NMS-Ag nanocomposite (NMSA6). This substrate greatly enhances the Raman signal, achieving an enhancement factor of approximately 1.49 × 107 and a detection limit as low as 10-11 M for simultaneous multi-component analysis. Finite-difference time-domain (FDTD) simulations confirm the enhancement mechanism. The NMSA6 substrate demonstrates remarkably low detection limits for BPA and melamine, facilitating the analysis of various hazardous substances. These findings highlight the substrate's potential for highly sensitive, label-free detection, presenting a viable tool for food safety monitoring.

IRTIDP: A simple integrated real-time isolation and detection platform for small extracellular vesicles Glypican-1 in pancreatic cancer patients.

Glypican-1 (GPC-1) protein-positive small extracellular vesicles (GPC-1+-sEV) have been proposed as potential biomarkers for early diagnosis of pancreatic cancer. In this study, we present an integrated real-time isolation and detection platform (IRTIDP) to capture and analyze GPC-1+-sEV directly from sera of pancreatic cancer patients. First, CD63 antibody-modified metal-organic framework (MOF) materials were utilized to enrich sEVs with a capture efficiency of 93.93 %. Second, a SERS probe was constructed by Raman reporter 4-MBA and GPC-1 antibody modified SERS active silver nanoparticles (AgNPs), which formed a sandwich complex structure of "MOFs@GPC-1+-sEV@AgNPs-4-MBA" with MOFs-enriched sEVs. The IRTSDP can complete the capture and detection process within 35 min, with a detection limit for 1 GPC-1+-sEV/µL, and linear range between 105∼109 GPC-1+-sEV/mL. Furthermore, this approach has been applied to quantify serum sEV GPC-1 in clinical pancreatic cancer patients. Based on the SERS intensity analysis, pancreatic cancer patients can be distinguished from pancreatic cystadenoma patients and healthy individuals effectively using this innovative platform that provides highly specific and sensitive means for early diagnosis of pancreatic cancer as well as other tumor types.

Highly sensitive SERS nanoplatform based on aptamer and vancomycin for detection of S. aureus and its clinical application.

Staphylococcus aureus (S. aureus) is the most common pathogen in human purulent infections, which can cause local purulent infections, as well as pneumonia, pseudomembranous enteritis, pericarditis, and even systemic infections. The conventional methods including bacteria colony counting, polymerase chain reaction and enzyme-linked immunosorbent assay can't fully meet the requirement of highly sensitive detection of S. aureus due to their own disadvantages. Therefore, it's an urgent need to develop new platform to detect S. aureus in the early infection stage. In this study, a new surface-enhanced Raman scattering (SERS)-based nanoplatform based on dual-recognition of aptamer (Apt) and vancomycin (Van) was developed for the highly sensitive detection of S. aureus. The SERS nanoplatform consisted of two functional parts: aptamer-conjugated Fe3O4 magnetic nanoparticles (Fe3O4-Apt MNPs) for bacteria enrichment and vancomycin modified-Au nanoparticles (Van-Au NPs) as the SERS probes for S. aureus quantitative detection. Upon the target bacteria enrichment, the SERS signals of the supernatant after magnetic separation could be obtained and analyzed under different concentrations of S. aureus. The limit of detection of the proposed assay was found to be 3.27 CFU/mL. We believe that the proposed SERS-based nanoplatform has great potential as a powerful tool in the early detection of specific bacteria.

DNA Origami-Engineered Plasmonic Nanoprobes for Targeted Cancer Imaging.

Plasmonic nanomaterials bearing targeting ligands are of great interest for surface-enhanced Raman scattering (SERS)-based bioimaging applications. However, the practical utility of SERS-based imaging strategies has been hindered by the lack of a straightforward method to synthesize highly sensitive SERS-active nanostructures with high yield and efficiency. In this work, leveraging DNA origami principles, we report the first-in-class design of a SERS-based plasmonically coupled nanoprobe for targeted cancer imaging (SPECTRA). The nanoprobe harnesses a cancer cell targeting DNA aptamer sequence and vibrational tag with stretching frequency in the cell-silent Raman window. Through the integration of aptamer sequence specific for DU145 cells, we show the unique capabilities of SPECTRA for targeted imaging of DU145 cells. Our results demonstrate that the scalability, cost-effectiveness, and reproducibility of this method of fabrication of SERS nanoprobes can serve as a versatile platform for creating nanoprobes with broad applications in the fields of cancer biology and biomedical imaging.

Surface Enhanced Nonlinear Raman Processes for Advanced Vibrational Probing.

Surface enhanced Raman scattering (SERS) is not restricted to the well-known one-photon excited spontaneous Raman process that gives information on molecular composition, structure, and interaction through vibrational probing with high sensitivity. The enhancement mainly originates in high local fields, specifically those provided by localized surface plasmon resonances of metal nanostructures. High local fields can particularly support nonlinear Raman scattering, as it depends on the fields to higher powers. By revealing plasmon-molecule interactions, nonlinear Raman processes provide a very sensitive access to the properties of metal nanomaterials and their interfaces with molecules and other materials. This Perspective discusses plasmon-enhanced spontaneous and coherent nonlinear Raman scattering with the aim of identifying advantages that lead to an advanced vibrational characterization of such systems. The discussion will highlight the aspects of vibrational information that can be gained based on specific advantages of different incoherent and coherent Raman scattering and their surface enhancement. While the incoherent process of surface enhanced hyper Raman scattering (SEHRS) gives highly selective and spectral information complementary to SERS, the incoherent process of surface enhanced pumped anti-Stokes Raman scattering (SEPARS) can help to infer effective nonresonant SERS cross sections and allows to see "hot" vibrational transitions. Surface enhanced coherent anti-Stokes Raman scattering (SECARS) and surface enhanced stimulated Raman scattering (SESRS) combine the advantages of high local fields and coherence, which gives rise to high detection sensitivity and offers possibilities to explore molecule-plasmon interactions for a comprehensive characterization of composite and hybrid structures in materials research, catalysis, and nanobiophotonics.

Unveiling the Mechanism of Plasmon Photocatalysis via Multiquantum Vibrational Excitation.

Plasmon photocatalysis reactions are thought to occur through vibrationally activated reactants, driven by nonthermal energy transfer from plasmon-induced hot carriers. However, a detailed quantum-state-level understanding and quantification of the activation have been lacking. Using anti-Stokes surface-enhanced Raman scattering (SERS) spectroscopy, we mapped the vibrational population distributions of reactants on plasmon-excited nanostructures. Our results reveal a highly nonthermal distribution with an anomalously enhanced population of multiquantum excited states (v ≥ 2). The shape of the distribution and its dependence on local field intensity and excitation wavelength cannot be explained by photothermal heating or vibronic optical transitions of the metal-molecule complex. Instead, it can be modeled by hot electron-molecule energy transfer mediated by the transient negative ions, establishing direct links among nonthermal reactant activation, plasmon-induced hot electrons, and negative ion resonances. Moreover, the presence of multiquantum excited reactants, which are far more reactive than those in the ground state or first excited state, presents opportunities for vibrationally controlling chemical selectivities.

Au@CuS Nanoshells for Surface-Enhanced Raman Scattering Image-Guided Tumor Photothermal Therapy with Accelerated Hepatobiliary Excretion.

Gold-based nanoparticles for surface-enhanced Raman scattering (SERS) imaging show great potential for precise tumor detection and photothermal therapy (PTT). However, the metabolizability of gold nanoparticles (Au NPs) raises big concerns. Herein, we designed a core-shelled nanostructure of copper sulfide (CuS)-coated Au NPs with surface pegylation (PEG-Au@CuS NSs). The excreted Au in the gallbladders at 1 h and 4 h in mice injected with PEG-Au@CuS NSs was 8.2- and 19.1-fold of that with the pegylated Au NPs (PEG-AuNPs) of the same Au particle size, respectively. By loading the Raman reporter 3,3'-Diethylthiatricarbocyanine iodide (DTTC) in the core-shell junction of PEG-Au@CuS NSs, the PEG-Au-DTTC@CuS NSs exhibited the Raman signal-to-noise (S/N) ratio of 4.01 after 24 h of intravenous (IV) injection in the mice bearing an orthotopic CT26-Luc colon tumor. By contrast, the DTTC-coated PEG-AuNPs (PEG-Au-DTTC NPs) achieved an S/N ratio of 2.71. Moreover, PEG-Au-DTTC@CuS NSs exhibited an increased photothermal conversion effect compared with PEG-Au-DTTC NPs excited with an 808-nm laser. PEG-Au-DTTC@CuS NSs enabled intraoperative SERS image-guided photothermal therapy for a complete cure of the colon tumor-bearing mice. Our data demonstrated that the PEG-Au-DTTC@CuS NSs are promising intraoperative Raman image-guided theranostic nanoplatform with enhanced hepatobiliary excretion.

Patterned Gold Nanoparticle Superlattice Film for Wearable Sweat Sensors.

Nanoparticle superlattices are beneficial in terms of providing strong and uniform signals in analysis owing to their closely packed uniform structures. However, nanoparticle superlattices are prone to cracking during physical activities because of stress concentrations, which hinders their detection performance and limits their analytical applications. In this work, template printing methods were used in this study to prepare a patterned gold nanoparticle (AuNP) superlattice film. By adjustment of the size of the AuNP superlattice domain below the critical size of fracture, the mechanical stability of the AuNP superlattice domain is improved. Thus, long-term sustainable high-performance signal output is achieved. The patterned AuNP superlattice film was used to construct a wearable sweat sensor based on surface-enhanced Raman scattering (SERS). The designed sensor showed promise for long-term reliable use in actual scenarios in terms of recommending water replenishment, monitoring hydration states, and tracking the intensity of activity.

One-step synthesized multisize AuAg alloy nanoparticles with high SERS sensitivity in directly detecting SARS-CoV-2 spike protein.

The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in widespread disease transmission, challenging the stability of global healthcare systems. Surface-enhanced Raman scattering (SERS) as an easy operation, fast, and low-cost technology illustrates a good potential in detecting SARS-CoV-2. In the study, one-step fabrication of gold-silver alloy nanoparticles (AuAgNPs) with adjustable metal proportions and diameters is employed as SERS substrates. The angiotensin-converting enzyme 2 (ACE2) functionalized AuAgNPs are applied as sensor surfaces to detect SARS-CoV-2 S protein. By optimizing the SERS substrates, ACE2/Au35Ag65NPs illustrate higher performance in detecting the SARS-CoV-2 S protein with a limit of detection (LOD) of 10 fg/mL in both phosphate-buffered saline (PBS) and pharyngeal swabs solution (PSS). It also provides excellent reproducibility with a relative standard deviation (RSD) of 7.7 % and 7.9 %, respectively. This easily preparable and highly reproducible SERS substrate has good potential in the practical application of detecting SARS-CoV-2.

Study of two-dimensional information writing, reading and error correction at micro/nanoscale based on gold nanosphere arrays.

Surface plasmon driven photocatalytic reactions have great potential for information encryption as well as information security. In this paper, explored the detection concentrations of dye molecule Rhodamine6G (R6G) on three substrates, where complete original Raman spectra signals were still obtained at a concentration of 10-8 M. Utilized photosensitive molecules to investigate the photocatalytic characteristics of 4-nitrobenzenethiol (4-NBT) on three substrates. Excitation light at a wavelength of 633 nm enables local photocatalytic for information signals writing, while 785 nm wavelength excitation light combined with two-dimensional Mapping technology is used for information signal reading. Read information signals are often prone to reading errors due to their own lack of resolution or strong interference from back bottom signals, so error correction processing of information signals is essential. Through comparative exploration, it is found that the ratio method can obtain high-precision and high-resolution information signals, and the interference of the background signals were well suppressed. Leveraging the advantages of Raman fingerprint spectra at the micro/nanoscale, it solves the challenge of incomplete information signals presentation at smaller scales. Additionally, through error correction processing of the information signals, high precision and high-resolution information signals are obtained.

SERS biosensors based on catalytic hairpin self-assembly and hybridization chain reaction cascade signal amplification strategies for ultrasensitive microRNA-21 detection.

A highly sensitive surface-enhanced Raman scattering (SERS) biosensor has been developed for the detection of microRNA-21 (miR-21) using an isothermal enzyme-free cascade amplification method involving catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR). The CHA reaction is triggered by the target miR-21, which causes hairpin DNA (C1 and C2) to self-assemble into CHA products. After AgNPs@Capture captures the resulting CHA product, the HCR reaction is started, forming long-stranded DNA on the surface of AgNPs. A strong SERS signal is generated due to the presence of a large amount of the Raman reporter methylene blue (MB) in the vicinity of the SERS "hot spot" on the surface of AgNPs. The monitoring of the SERS signal changes of MB allows for the highly sensitive and specific detection of miR-21. In optimal conditions, the biosensor exhibits a satisfactory linear range and a low detection limit for miR-21 of 42.3 fM. Additionally, this SERS biosensor shows outstanding selectivity and reproducibility. The application of this methodology to clinical blood samples allows for the differentiation of cancer patients from healthy controls. As a result, the CHA-HCR amplification strategy used in this SERS biosensor could be a useful tool for miRNA detection and early cancer screening.

Portable point-of-care surface enhanced Raman scattering spectroscopy for the quantification of glutathione in whole blood microsamples.

Glutathione (GSH) is a non-protein tripeptide thiol that plays a prominent role in oxidative stress defense. GSH concentration is particularly critical in the neonatal period, especially for premature newborns that face increased susceptibility to oxidative stress. Monitoring GSH levels provides valuable insights into newborn health, helping to tailor care to their specific needs. The aim of this study was the development of a sensor specifically targeted for its use in neonatology, enabling GSH determination in only 2 µL of whole blood. The newly developed sensing system simplifies sample processing, addressing a critical need in clinical applications. Unlike current methods that demand fast pre-processing of relatively large sample volumes, expensive equipment, and skilled personnel, the developed approach streamlines the analytical process. By using 2 µL of whole blood, a single syringe filter for sample treatment, a deuterated internal standard (IS) for signal normalization, and Surface Enhanced Raman Scattering (SERS) spectroscopy with a silver colloid substrate for GSH detection, the set-up's characteristics are compatible with point-of-care applications. The analytical procedure was validated and applied to diverse populations including healthy adults (N = 63) and newborns (N = 35), yielding GSH concentration values ranging from 0.6 to 1.8 and 0.8-2.1 mM, respectively. This new optical sensor offers a quick and cost-effective solution to support the assessment of GSH levels in newborns that can greatly benefit not only neonatal care, but also the study of adult populations for health monitoring.

Optimisation of a Microwave Synthesis of Silver Nanoparticles by a Quality by Design Approach to Improve SERS Analytical Performances.

A major limitation preventing the use of surface-enhanced Raman scattering (SERS) in routine analyses is the signal variability due to the heterogeneity of metallic nanoparticles used as SERS substrates. This study aimed to robustly optimise a synthesis process of silver nanoparticles to improve the measured SERS signal repeatability and the protocol synthesis repeatability. The process is inspired by a chemical reduction method associated with microwave irradiation to guarantee better controlled and uniform heating. The innovative Quality by Design strategy was implemented to optimise the different parameters of the process. A preliminary investigation design was firstly carried out to evaluate the influence of four parameters selected by means of an Ishikawa diagram. The critical quality attributes were to maximise the intensity of the SERS response and minimise its variance. The reaction time, temperature and stirring speed are critical process parameters. These were optimised using an I-optimal design. A robust operating zone covering the optimal reaction conditions (3.36 min-130 °C-600 rpm) associated with a probability of success was modelled. Validation of this point confirmed the prediction with intra- and inter-batch variabilities of less than 15%. In conclusion, this study successfully optimised silver nanoparticles by a rapid, low cost and simple technique enhancing the quantitative perspectives of SERS.

Generic method for the detection of short & long chain PFAS extended to the lowest concentration levels of SERS capability.

The detection of the highly toxic per- and polyfluoroalkyl substances, PFAS, constitutes a challenging task in terms of developing a generic method that could be rapid and applicable simultaneously to both long and short-chain PFAS at ppt concentration level. In the present study, the method introduced by the USA Environmental Protection Agency, EPA, to detect surfactants, using methylene blue, MB, which is identified an ideal candidate for PFAS-MB ion pairing, is extended at the lowest concentration range by a simple additional step that involves the dissociation of the ion pairs in water. In this work, Surface Enhanced Raman Scattering, SERS, is applied via Ag nanocolloidal suspensions to probe MB and indirectly either/or both short-chain (perfluorobutyric acid, PFBA) and long-chain (perfluoloctanoic acid, PFOA) PFAS downt to 5 ppt. This method, which can be further optimized to sub-ppt level via a custom-made SERS-PFAS dedicated Raman system, offers the possibility to be applied to either specific PFAS (both short and long-chain) in a targeted analysis or to total PFAS in a non-targeted analysis at very low detection limits, following any type of MB detection method in aqueous solutions and obviously with any type of SERS substrate.

Combined Electrochemical Deposition and Photo-Reduction to Fabricate SERS-Active Silver Substrates: Characterization and Application for Malachite Green Detection in Aquaculture Water.

This research introduces a novel approach using silver (Ag) nanostructures generated through electrochemical deposition and photo-reduction of Ag on fluorine-doped tin oxide glass substrates (denoted as X-Ag-AgyFTO, where 'X' and 'y' represent the type of light source and number of deposited cycles, respectively) for surface-enhanced Raman spectroscopy (SERS). This study used malachite green (MG) as a Raman probe to evaluate the enhancement factors (EFs) in SERS-active substrates under varied fabrication conditions. For the substrates produced via electrochemical deposition, we determined a Raman EF of 6.15 × 104 for the Ag2FTO substrate. In photo-reduction, the impact of reductant concentration, light source, and light exposure duration were examined on X-Ag nanoparticle formation to achieve superior Raman EFs. Under optimal conditions (9.0 mM sodium citrate, 460 nm blue-LED at 10 W for 90 min), the combination of blue-LED-reduced Ag (B-Ag) and an Ag2FTO substrate (denoted as B-Ag-Ag2FTO) exhibited the best Raman EF of 2.79 × 105. This substrate enabled MG detection within a linear range of 0.1 to 1.0 µM (R2 = 0.98) and a detection limit of 0.02 µM. Additionally, the spiked recoveries in aquaculture water samples were between 90.0% and 110.0%, with relative standard deviations between 3.9% and 6.3%, indicating the substrate's potential for fungicide detection in aquaculture.

In Situ Collection and SERS Detection of Nitrite in Exhalations on Facemasks Based on Wettability Differences.

As one of the common carriers of biological information, along with human urine specimens and blood, exhaled breath condensate (EBC) carries reliable and rich information about the body's metabolism to track human physiological normal/abnormal states and environmental exposures. What is more, EBC has gained extensive attention because of the convenient and nondestructive sampling. Facemasks, which act as a physical filter barrier between human exhaled breath and inhaled substances from the external environment, are safe, noninvasive, and economic devices for direct sampling of human exhaled breath and inhaled substances. Inspired by the ability of fog collection of Namib desert beetle, a strategy for in situ collecting and detecting EBC with surface-enhanced Raman scattering is illustrated. Based on the intrinsic and unique wettability differences between the squares and the surrounding area of the pattern on facemasks, the hydrophilic squares can capture exhaled droplets and spontaneously enrich the analytes and silver nanocubes (AgNCs), resulting in good repeatability in situ detection. Using R6G as the probe molecule, the minimal detectable concentration can reach as low as 10-16 M, and the relative standard deviation is less than 7%. This proves that this strategy can achieve high detection sensitivity and high detection repeatability. Meanwhile, this strategy is applicable for portable nitrite analysis in EBC and may provide an inspiration for monitoring other biomarkers in EBC.