Yolk-Shell Hierarchical Pore Au@MOF Nanostructures: Efficient Gas Capture and Enrichment for Advanced Breath Analysis.

Surface-enhanced Raman scattering (SERS) offers a promising, cost-effective alternative for the rapid, sensitive, and quantitative analysis of potential biomarkers in exhaled gases, which is crucial for early disease diagnosis. However, a major challenge in SERS is the effective detection of gaseous analytes, primarily due to difficulties in enriching and capturing them within the substrate's "hotspot" regions. This study introduces an advanced gas sensor combining mesoporous gold (MesoAu) and metal-organic frameworks (MOFs), exhibiting high sensitivity and rapid detection capabilities. The MesoAu provides abundant active sites and interconnected mesopores, facilitating the diffusion of analytes for detection. A ZIF-8 shell enveloping MesoAu further enriches target molecules, significantly enhancing sensitivity. A proof-of-concept experiment demonstrated a detection limit of 0.32 ppb for gaseous benzaldehyde, indicating promising prospects for the rapid diagnosis of early stage lung cancer. This research also pioneers a novel approach for constructing hierarchical plasmonic nanostructures with immense potential in gas sensing.

Plasmonic Dodecahedral-Walled Elongated Nanoframes for Surface-Enhanced Raman Spectroscopy.

Here, elongated pseudohollow nanoframes composed of four rectangular plates enclosing the sides and two open-frame ends with four ridges pointing at the tips for near-field focusing are reported. The side facets act as light-collecting domains and transfer the collected light to the sharp tips for near-field focusing. The nanoframes are hollow inside, allowing the gaseous analyte to penetrate through the entire architecture and enabling efficient detection of gaseous analytes when combined with Raman spectroscopy. The resulting nanostructures are named Au dodecahedral-walled nanoframes. Synthesis of the nanoframes involves shape transformation of Au nanorods with round tips to produce Au-elongated dodecahedra, followed by facet-selective Pt growth, etching of the inner Au, and regrowth steps. The close-packed assembly of Au dodecahedral-walled nanoframes exhibits an attomolar limit of detection toward benzenethiol. This significant enhancement in SERS is attributed to the presence of a flat solid terrace for a large surface area, sharp edges and vertices for strong electromagnetic near-field collection, and open frames for effective analyte transport and capture. Moreover, nanoframes are applied to detect chemical warfare agents, specifically mustard gas simulants, and 20 times higher sensitivity is achieved compared to their solid counterparts.

Speciation Analysis of Metals and Metalloids by Surface Enhanced Raman Spectroscopy.

The presence of metalloids and heavy metals in the environment is of critical concern due to their toxicological impacts. However, not all metallic species have the same risk level. Specifically, the physical, chemical, and isotopic speciation of the metal(loids) dictate their metabolism, toxicity, and environmental fate. As such, speciation analysis is critical for environmental monitoring and risk assessment. In the past two decades, surface-enhanced Raman spectroscopy (SERS) has seen significant developments regarding trace metal(loid) sensing due to its ultrahigh sensitivity, readiness for in situ real-time applications, and cost-effectiveness. However, the speciation of metal(loid)s has not been accounted for in the design and application of SERS sensors. In this Perspective, we examine the potential of SERS for metal(loid) speciation analysis and highlight the advantages, progress, opportunities, and challenges of this application.

Analysis and experimental assessment of an optimized SERS substrate used to detect thiabendazole in apples with high sensitivity.

Pesticides that linger in the environment and ecosystems for an extended period can cause severe and dangerous health problems in humans. To detect pesticides in foods, the development of high-sensitivity and quick screening technologies was required. This research investigated the performance of Au@Ag NPs with varying thicknesses of the silver shell for detecting trace quantities of thiabendazole (TBZ) in apples using surface-enhanced Raman spectroscopy (SERS). The Au@Ag NPs were synthesized by coating 32 nm gold seeds with different thicknesses of silver shell ranging from 2.4 to 8.7 nm, achieved by adjusting the incorporation of AgNO3 and ascorbic acid. The optimized Au@Ag NPs with a 7.3 nm silver shell demonstrated outstanding SERS activity, high sensitivity, and a detection limit of 0.05 µg/mL for TBZ. The R2 values, representing the goodness of fit, were found to be 0.990 and 0.986 for standard and real TBZ samples, respectively, indicating a strong correlation between the measured signal and the TBZ concentration. The recovery analysis showed a reliable and accurate detection capability (96 to 105%), suggesting good reliability and accuracy of the SERS-based detection using the optimal Au@Ag NPs. Overall, this research highlights the potential of SERS with optimal Au@Ag NPs for rapid and effective monitoring of pesticides in the food industry.

Rapid detection of African swine fever virus via SERS probe-modified sandwich hybridization assay.

Rapid and reliable detection method for African swine fever virus (ASFV) is proposed by surface-enhanced Raman spectroscopy (SERS). The ASFV target DNA can be specifically captured by sandwich hybridization between nanomagnetic beads and a SERS probe. Experimental results show that the significant Raman signal of the SERS probe with gold nanoparticles and a molecular reporter DTNB (5,5'-dimercapto-bis (2-nitrobenzoic acid)) can be adopted for detecting the hybridization chain reaction of ASFV DNA. The advantage of the SERS sandwich hybridization assay is the large response range from the single molecule level to 108 copies per mL, which not only can overcome the tedious time required for the amplification reaction but also provides a comparative method to polymerase chain reaction. Furthermore, real samples of African swine fever virus were detected from different subjects of swine fever virus including porcine reproductive respiratory syndrome virus and Japanese encephalitis virus. The proposed biosensor method can rapidly detect ASFV correctly within 15 min as a simple, convenient, low-cost detection approach. The biosensor can be used as a platform for the determination in biological, food, and environmental analytical fields.

Facile Preparation of TiO2NTs/Au@MOF Nanocomposites for High-Sensitivity SERS Sensing of Gaseous VOC.

Surface-enhanced Raman spectroscopy (SERS) is a promising and highly sensitive molecular fingerprint detection technology. However, the development of SERS nanocomposites that are label-free, highly sensitive, selective, stable, and reusable for gaseous volatile organic compounds (VOCs) detection remains a challenge. Here, we report a novel TiO2NTs/AuNPs@ZIF-8 nanocomposite for the ultrasensitive SERS detection of VOCs. The three-dimensional TiO2 nanotube structure with a large specific surface area provides abundant sites for the loading of Au NPs, which possess excellent local surface plasmon resonance (LSPR) effects, further leading to the formation of a large number of SERS active hotspots. The externally wrapped porous MOF structure adsorbs more gaseous VOC molecules onto the noble metal surface. Under the synergistic mechanism of physical and chemical enhancement, a better SERS enhancement effect can be achieved. By optimizing experimental conditions, the SERS detection limit for acetophenone, a common exhaled VOC, is as low as 10-11 M. And the relative standard deviation of SERS signal intensity from different points on the same nanocomposite surface is 4.7%. The acetophenone gas achieves a 1 min response and the signal reaches stability in 4 min. Under UV irradiation, the surface-adsorbed acetophenone can be completely degraded within 40 min. The experimental results demonstrate that this nanocomposite has good detection sensitivity, repeatability, selectivity, response speed, and reusability, making it a promising sensor for gaseous VOCs.

Hot Spot Engineering in Hierarchical Plasmonic Nanostructures.

The controllable nanogap structures offer an effective way to obtain strong and tunable localized surface plasmon resonance (LSPR). A novel hierarchical plasmonic nanostructure (HPN) is created by incorporating a rotating coordinate system into colloidal lithography. In this nanostructure, the hot spot density is increased drastically by the long-range ordered morphology with discrete metal islands filled in the structural units. Based on the Volmer-Weber growth theory, the precise HPN growth model is established, which guides the hot spot engineering for improved LSPR tunability and strong field enhancement. The hot spot engineering strategy is examined by the application of HPNs as the surface-enhanced Raman spectroscopy (SERS) substrate. It is universally suitable for various SERS characterization excited at different wavelengths. Based on the HPN and hot spot engineering strategy, single-molecule level detection and long-range mapping can be realized simultaneously. In that sense, it offers a great platform and guides the future design for various LSPR applications like surface-enhanced spectra, biosensing, and photocatalysis.

Ultrasensitive SERS substrate for label-free therapeutic drug monitoring of chlorpromazine hydrochloride and aminophylline in human serum.

Surface-enhanced Raman spectroscopy (SERS) has been widely used in the field of therapeutic drug monitoring (TDM) because of its powerful fingerprinting capability. In this paper, we used an in situ synthesis method to anchor Ag nanoparticles (AgNPs) on the surface of MIL-101(Cr) to obtain MIL-101(Cr)@Ag. Owing to the large specific surface area and ultra-high porosity of MIL-101(Cr)@Ag, we developed a method for the determination of chlorpromazine hydrochloride (CPZ) and aminophylline (AMP) in human serum by using it as a solid-phase extraction sorbent and SERS substrate. The label-free TDM-SERS method was able to evaluate the levels of CPZ and AMP in serum samples with detection limits as low as 8.91 × 10-2 µg/mL and 3.4 × 10-2 µg/mL, respectively. In addition, influencing factors including sample solution pH, AgNO3 concentration, drug adsorption time, and the amount of sample solution were optimized. This protocol provides a new method with good selectivity, stability, reproducibility, homogeneity, and sensitivity for the determination of small-molecule drug content in serum samples. This label-free TDM-SERS method will help to achieve rapid individualized dosing regimens in clinical practice and has potential applications in the field of TDM.

Preparation of SERS base membrane with cellulose compound dopamine and determination of hypochlorite.

Flexible silver substrates were made by in situ reduction of silver nanoparticles in bacterial cellulose membranes using the unique advantage of dopamine. Subsequently, we modified the substrate with 4-mercaptophenol (4-MP), a molecule capable of specifically recognizing ClO-, and its corresponding SERS signal changes with the concentration of hypochlorite, thus allowing the quantitative detection of ClO- content. The method showed a negative linear correlation (R2 = 0.9567) with the SERS intensity at 1077 cm-1 over the concentration range 0.5-100 µM, and the detection limit was 0.15 µM. The RSD of the SERS intensity at 1077 cm-1 under five batches was 4.2%, which proved the good reproducibility of P-BCM-Ag NP-MP. Finally, the P-BCM-Ag NPs were used for the detection of hypochlorite in cell contents, artificial urine, and clinical serum samples, utilizing spike experiments in all three environments. The recoveries were in the range 90-110% indicating the accuracy of the method for the detection of hypochlorite and validating the promising application of this assay for practical detection in intricate biological samples.

A novel "on-off" SERS nanoprobe based on sulfonated cellulose nanofiber-Ag composite for selective determination of NADH in human serum.

A novel S-CNF-based nanocomposite was created using sulfonated cellulose nanofiber (S-CNF) to enable the detection of NADH in serum by surface-enhanced Raman spectroscopy (SERS). The numerous hydroxyl and sulfonic acid groups on the S-CNF surface absorbed silver ions and converted them to silver seeds, which formed the load fulcrum. After adding a reducing agent, silver nanoparticles (Ag NPs) were firmly adhered to the S-CNF surface to form stable 1D "hot spots." The S-CNF-Ag NP substrate demonstrated outstanding SERS performance, including good uniformity with an RSD of 6.88% and an enhancement factor (EF) of 1.23 × 107. Owing to the anionic charge repulsion effect, the S-CNF-Ag NP substrate still maintains remarkable dispersion stability after 12 months of preservation. Finally, S-CNF-Ag NPs' surface was modified with 4-mercaptophenol (4-MP), a special redox Raman signal molecule, to detect reduced nicotinamide adenine dinucleotide (NADH). The results showed that the detection limit (LOD) of NADH was 0.75 µM; a good linear relationship (R2 = 0.993) was established in the concentration range 10-6 - 10-2 M. The SERS nanoprobe enabled rapid detection of NADH in human serum without any complicated sample pretreatment and provides a new potential to detect biomarkers.

Raman-Based Techniques in Medical Applications for Diagnostic Tasks: A Review.

Raman spectroscopy is a widely developing approach for noninvasive analysis that can provide information on chemical composition and molecular structure. High chemical specificity calls for developing different medical diagnostic applications based on Raman spectroscopy. This review focuses on the Raman-based techniques used in medical diagnostics and provides an overview of such techniques, possible areas of their application, and current limitations. We have reviewed recent studies proposing conventional Raman spectroscopy and surface-enhanced Raman spectroscopy for rapid measuring of specific biomarkers of such diseases as cardiovascular disease, cancer, neurogenerative disease, and coronavirus disease (COVID-19). As a result, we have discovered several most promising Raman-based applications to identify affected persons by detecting some significant spectral features. We have analyzed these approaches in terms of their potentially diagnostic power and highlighted the remaining challenges and limitations preventing their translation into clinical settings.

Advances in surface-enhanced Raman spectroscopy technology for detection of foodborne pathogens.

Rapid control and prevention of diseases caused by foodborne pathogens is one of the existing food safety regulatory issues faced by various countries and has received wide attention from all sectors of society. The development of rapid and reliable detection methods for foodborne pathogens remains a hot research area for food safety and public health because of the limitations of complex steps, time-consuming, low sensitivity, or poor selectivity of commonly used methods. Surface-enhanced Raman spectroscopy (SERS), as a novel spectroscopic technique, has the advantages of high sensitivity, selectivity, rapid and nondestructive detection and has exhibited broad application prospects in the determination of pathogenic bacteria. In this study, the enhancement mechanisms of SERS are briefly introduced, then the characteristics and properties of liquid-phase, rigid solid-phase, and flexible solid-phase are categorized. Furthermore, a comprehensive review of the advances in label-free or label-based SERS strategies and SERS-compatible techniques for the detection of foodborne pathogens is provided, and the advantages and disadvantages of these methods are reviewed. Finally, the current challenges of SERS technology applied in practical applications are listed, and the possible development trends of SERS in the field of foodborne pathogens detection in the future are discussed.

Superhydrophobic 3D-Assembled Metallic Nanoparticles for Trace Chemical Enrichment in SERS Sensing.

The performance of surface-enhanced Raman spectroscopy (SERS) is determined by the interaction between highly diluted analytes and boosted localized electromagnetic fields in nanovolumes. Although superhydrophobic surfaces are developed for analyte enrichment, i.e., to concentrate and transfer analytes toward a specific position, it is still challenging to realize reproducible, uniform, and sensitive superhydrophobic SERS substrates over large scales, representing a major barrier for practical sensing applications. To overcome this challenge, a superhydrophobic SERS chip that combines 3D-assembled gold nanoparticles on nanoporous substrates is proposed, for a strong localized field, with superhydrophobic surface treatment for analyte enrichment. Intriguingly, by concentrating droplets in the volume of 40 µL, the sensitivity of 1 nm is demonstrated using 1,2-bis(4-pyridyl)-ethylene molecules. In addition, this unique chip demonstrates a relative standard deviation (RSD) of 2.2% in chip-to-chip reproducibility for detection of fentanyl at 1 µg mL-1 concentration, revealing its potential for quantitative sensing of chemicals and drugs. Furthermore, the trace analysis of fentanyl and fentanyl-heroin mixture in human saliva is realized after a simple pretreatment process. This superhydrophobic chip paves the way toward on-site and real-time drug sensing to tackle many societal issues like drug abuse and the opioid crisis.

Biomimetic Transparent Nanoplasmonic Meshes by Reverse-Nanoimprinting for Bio-Interfaced Spatiotemporal Multimodal SERS Bioanalysis.

Multicellular systems, such as microbial biofilms and cancerous tumors, feature complex biological activities coordinated by cellular interactions mediated via different signaling and regulatory pathways, which are intrinsically heterogeneous, dynamic, and adaptive. However, due to their invasiveness or their inability to interface with native cellular networks, standard bioanalysis methods do not allow in situ spatiotemporal biochemical monitoring of multicellular systems to capture holistic spatiotemporal pictures of systems-level biology. Here, a high-throughput reverse nanoimprint lithography approach is reported to create biomimetic transparent nanoplasmonic microporous mesh (BTNMM) devices with ultrathin flexible microporous structures for spatiotemporal multimodal surface-enhanced Raman spectroscopy (SERS) measurements at the bio-interface. It is demonstrated that BTNMMs, supporting uniform and ultrasensitive SERS hotspots, can simultaneously enable spatiotemporal multimodal SERS measurements for targeted pH sensing and non-targeted molecular detection to resolve the diffusion dynamics for pH, adenine, and Rhodamine 6G molecules in agarose gel. Moreover, it is demonstrated that BTNMMs can act as multifunctional bio-interfaced SERS sensors to conduct in situ spatiotemporal pH mapping and molecular profiling of Escherichia coli biofilms. It is envisioned that the ultrasensitive multimodal SERS capability, transport permeability, and biomechanical compatibility of the BTNMMs can open exciting avenues for bio-interfaced multifunctional sensing applications both in vitro and in vivo.

Targeted Suppression of Peptide Degradation in Ag-Based Surface-Enhanced Raman Spectra by Depletion of Hot Carriers.

Sample degradation, in particular of biomolecules, frequently occurs in surface-enhanced Raman spectroscopy (SERS) utilizing supported silver SERS substrates. Currently, thermal and/or photocatalytic effects are considered to cause sample degradation. This paper establishes the efficient inhibition of sample degradation using iodide which is demonstrated by a systematic SERS study of a small peptide in aqueous solution. Remarkably, a distinct charge separation-induced surface potential difference is observed for SERS substrates under laser irradiation using Kelvin probe force microscopy. This directly unveils the photocatalytic effect of Ag-SERS substrates. Based on the presented results, it is proposed that plasmonic photocatalysis dominates sample degradation in SERS experiments and the suppression of typical SERS sample degradation by iodide is discussed by means of the energy levels of the substrate under mild irradiation conditions. This approach paves the way toward more reliable and reproducible SERS studies of biomolecules under physiological conditions.

Photochemical synthesis of pink silver and its use for monitoring peptide nitration via surface enhanced Raman spectroscopy (SERS).

Oxidative stress may cause extended tyrosine posttranslational modifications of peptides and proteins. The 3-nitro-L-tyrosine (Nit), which is typically formed, affects protein behavior during neurodegenerative processes, such as Alzheimer's and Parkinson's diseases. Such metabolic products may be conveniently detected at very low concentrations by surface enhanced Raman spectroscopy (SERS). Previously, we have explored the SERS detection of the Nit NO2 bending vibrational bands in a presence of hydrogen chloride (Niederhafner et al., Amino Acids 53:517-532, 2021, ibid). In this article, we describe performance of a new SERS substrate, "pink silver", synthesized photochemically. It provides SERS even without the HCl induction, and the acid further decreases the detection limit about 9 times. Strong SERS bands were observed in the asymmetric (1550-1475 cm-1) and symmetric (1360-1290 cm-1) NO stretching in the NO2 group. The bending vibration was relatively weak, but appeared stronger when HCl was added. The band assignments were supported by density functional theory modeling.

Plasmonic features of free-standing chitosan nanocomposite film with silver and graphene oxide for SERS applications.

A scalable procedure of SERS substrates design was developed using a novel plasmonic structure based on a freestanding chitosan film, silver nanoparticles, and graphene oxide. Chitosan provides a uniform distribution of silver nanoparticles from a colloidal suspension and, therefore, a reproducible Raman signal from local areas of measurements of several tens of microns. The addition of graphene oxide (GO) to the colloidal solution of silver nanoparticles suppresses the tortuous background fluorescence signal from the analyte and leads to an increase in the signal-to-fluorescence background intensity ratio by up to 6 times as compared to structures without GO. The manufactured plasmonic polymer nanocomposite provides a detection limit of down to 100 pM for R6G using a laser wavelength of 532 nm through a portable ×10 objective. The high colloidal stability of GO in water and the use of an aqueous colloid of silver nanoparticles simplify the procedure for creating a substrate by applying the GO-silver composite on the surface of a chitosan film without a need to form a GO film. Therefore, our approach paves a promising avenue to provide more sensitive detection even for the fluorescent analytes with short-wavelength lasers (532, 633 nm) instead of IR (785, 1024 nm) and foster the practical application of the developed plasmonic composites on portable Raman spectrometers.

Ultrasensitive detection of plant hormone abscisic acid-based surface-enhanced Raman spectroscopy aptamer sensor.

Abscisic acid (ABA), as the most common plant hormone in the growth of wheat, can greatly affect the yield when its levels deviate from normal. Therefore, highly sensitive and selective detection of this hormone is greatly needed. In this work, we developed an aptamer sensor based on surface-enhanced Raman spectroscopy (SERS) and applied it for the high sensitivity detection of ABA. Biotin-modified ABA aptamer complement chains were modified on ferrosoferric oxide magnetic nanoparticles (Fe3O4MNPs) and acted as capture probes, and sulfhydryl aptamer (SH-Apt)-modified silver-coated gold nanospheres (Au@Ag NPs) were used as signal probes. Through the recognition of the ABA aptamer and its complementary chains, an aptamer sensor based on SERS was constructed. As SERS internal standard molecules of 4-mercaptobenzoic acid (4-MBA) were encapsulated between the gold core and silver shell of the signal probes; the constructed aptamer sensor generated a strong SERS signal of 4-MBA after magnetic separation. When there were ABA molecules in the detection system, with the preferential binding of ABA aptamer and ABA molecule, the signal probes were released from the capture probes, after magnetic separation, leading to a linear decrease in SERS intensity of 4-MBA. Thus, the detection response was linear over a logarithmic concentration range, with an ultra-low detection limit of 0.67 fM. In addition, the practical use of this assay method was demonstrated in ABA detection from fresh wheat leaves, with a relative error (RE) of 5.43-8.94% when compared with results from enzyme-linked immunosorbent assay (ELISA). The low RE value proves that the aptamer sensor will be a promising method for ABA detection.

Fabrication of a surface-enhanced Raman spectroscopy-based analytical method consisting of multifunctional DNA three-way junction-conjugated porous gold nanoparticles and Au-Te nanoworm for C-reactive protein detection.

C-Reactive protein (CRP) is a biomarker of inflammatory responses and an index for assessing the risk of cardiovascular disease and estimating prognosis. In this study, we constructed a surface-enhanced Raman spectroscopy (SERS) biosensor composed of a multifunctional DNA three-way junction (DNA 3WJ), porous gold nanoplates (pAuNPs), and an Au-Te nanoworm structure for detection of CRP. The pAuNP and Au-Te nanostructures were synthesized by galvanic replacement reactions, and the morphology was confirmed by transmission electron microscopy, scanning electron microscopy, and dynamic light scattering (DLS). To generate the SERS signal, the Au-Te nanostructure was immobilized on an indium-tin oxide substrate, and the thiol-modified CRP aptamer was then self-assembled onto the modified substrate for CRP recognition. To amplify the SERS signal and identify the Raman tag, the multifunctional DNA 3WJ was conjugated with the pAuNPs, and each fragment of 3WJ was functionalized to biotin (pAuNP conjugation), methylene blue (Raman reporter), and CRP aptamer (target binding). The results were confirmed by gel electrophoresis. For conjugation between pAuNPs and DNA 3WJ, avidin was encapsulated in pAuNPs, and the conjugation structure was confirmed by DLS. The fabricated SERS biosensor showed detection limits of 2.23 pM in phosphate-buffered saline and 3.11 pM in diluted human serum. Overall, the proposed biosensor may have potential applications as a SERS biosensor platform.

Highly sensitive near-infrared SERS nanoprobes for in vivo imaging using gold-assembled silica nanoparticles with controllable nanogaps.

BACKGROUND: To take advantages, such as multiplex capacity, non-photobleaching property, and high sensitivity, of surface-enhanced Raman scattering (SERS)-based in vivo imaging, development of highly enhanced SERS nanoprobes in near-infrared (NIR) region is needed. A well-controlled morphology and biocompatibility are essential features of NIR SERS nanoprobes. Gold (Au)-assembled nanostructures with controllable nanogaps with highly enhanced SERS signals within multiple hotspots could be a breakthrough. RESULTS: Au-assembled silica (SiO2) nanoparticles (NPs) (SiO2@Au@Au NPs) as NIR SERS nanoprobes are synthesized using the seed-mediated growth method. SiO2@Au@Au NPs using six different sizes of Au NPs (SiO2@Au@Au50-SiO2@Au@Au500) were prepared by controlling the concentration of Au precursor in the growth step. The nanogaps between Au NPs on the SiO2 surface could be controlled from 4.16 to 0.98 nm by adjusting the concentration of Au precursor (hence increasing Au NP sizes), which resulted in the formation of effective SERS hotspots. SiO2@Au@Au500 NPs with a 0.98-nm gap showed a high SERS enhancement factor of approximately 3.8 × 106 under 785-nm photoexcitation. SiO2@Au@Au500 nanoprobes showed detectable in vivo SERS signals at a concentration of 16 µg/mL in animal tissue specimen at a depth of 7 mm. SiO2@Au@Au500 NPs with 14 different Raman label compounds exhibited distinct SERS signals upon subcutaneous injection into nude mice. CONCLUSIONS: SiO2@Au@Au NPs showed high potential for in vivo applications as multiplex nanoprobes with high SERS sensitivity in the NIR region.