Surface-enhanced Raman Scattering Nanotags Research Articles

In situ synthesis of gold nanoparticles on layered double hydroxide nanoparticles for multiplexing molecular imaging of single cells

Quasi-spherical gold nanoparticles (AuNPs) are the most widely used surface-enhanced Raman scattering (SERS) nanotags. However, their surface plasmon resonance properties are significantly restricted by their poor colloidal stability in biological media. Herein, we developed a new strategy for in situ synthesis of AuNPs on layered double hydroxide (LDH) nanoparticles to improve the colloidal stability of AuNPs for SERS imaging with multiplexing capability using single laser excitation. The hybridized LDH-AuNPs were constructed via in situ growth of AuNPs on lactic acid-LDH nanoparticles. The size of AuNPs was tuned by changing the concentration of LDH. The LDH-AuNPs were further functionalized with Raman reporter molecules (Ra) to obtain LDH-AuNPs-Ra as SERS nanotags, which were examined in terms of the colloidal stability and photostability in biological media as well as cellular uptake in J774A.1 macrophage cell culture by single cell mapping via confocal Raman microscopy. The cellular uptake of multiple LDH-AuNPs-Ra functionalized with different Raman reporter molecules were further investigated, where the multiplexing capability of SERS assay endows simultaneous detection of multiple LDH-AuNPs-Ra distribution after cellular uptake for cell imaging. In summary, the LDH-supported in situ synthesis of AuNPs have shown the merits of stabilizing AuNPs by using LDH for SERS imaging, as well as demonstrating the multiplexing molecular imaging of single cells. This work thus provides a promising strategy and a new bioimaging nanoplatform for various biomedical studies in the near future.

Multiplex signal amplification strategy-based early-stage diagnosis of Parkinson's disease on a SERS-enabled LoC system.

In this paper, a multiplex signal amplification strategy was developed for the determination of miR-214 and miR-221 on a surface-enhanced Raman scattering (SERS)-enabled lab-on-a-chip (LoC) system to realize the early-stage diagnosis of Parkinson's disease (PD). The gold nanobipyramids (GNBPs) with great monodispersity were functionalized with Raman reporter molecules and hairpin DNA 1, serving as the SERS nanotags. The presence of targets can initial the strand displacement amplification (SDA) reaction and the numerous short-stranded trigger DNA (tDNA) can be released under the action of polymerase and nicking enzyme. Then, the tDNA can trigger the catalytic hairpin assembly (CHA) event between the SERS nanotags and the capture nanoprobes (Magnetic beads (MBs) modified with hairpin DNA 2), resulting in the aggregation of GNBPs on the MBs surface. The multiplex signal amplification contributed by the SDA-CHA strategy and the magnet-induced aggregation effect can ultimately lead to the significant improvement of the detection sensitivity and the limit of detection (LOD) was low to aM level with reproducibility and specificity meanwhile. Furthermore, a MPTP-induced PD mice model was established to verify the practicability and the expression level of miR-214 and miR-221 at different stages analyzed with the LoC system was confirmed by qRT-PCR.

Engineering a SERS Sensing Nanoplatform with Self-Sterilization for Undifferentiated and Rapid Detection of Bacteria

The development of a convenient, sensitive, rapid and self-sterilizing biosensor for microbial detection is important for the prevention and control of foodborne diseases. Herein, we designed a surface-enhanced Raman scattering (SERS) sensing nanoplatform based on a capture-enrichment-enhancement strategy to detect bacteria. The gold-Azo@silver-cetyltrimethylammonium bromide (Au-Azo@Ag-CTAB) SERS nanotags were obtained by optimizing the synthesis process conditions. The results showed that the modification of CTAB enabled the nanotags to bind to different bacteria electrostatically. This SERS sensing nanoplatform was demonstrated to be fast (15 min), accurate and sensitive (limit of detection (LOD): 300 and 400 CFU/mL for E. coli and S. aureus, respectively). Of note, the excellent endogenous antibacterial activity of CTAB allowed the complete inactivation of bacteria after the assay process, thus effectively avoiding secondary contamination.

Reporter Molecules Embedded Au@Ag Core-Shell Nanospheres as SERS Nanotags for Cardiac Troponin I Detection

Rapid and accurate detection of acute myocardial infarction can improve patients' chances of survival. Cardiac troponin I (cTn I) is an important diagnostic biomarker for acute myocardial infarction. However, current immunoassays are insufficient to accurately measure cTn I, as they have limited detection sensitivity and are time-consuming. Surface-enhanced Raman scattering (SERS) is a brilliant fingerprints diagnostic technique characterised by ultrasensitivity, fast response, and qualitative and quantitative analysis capabilities. In this study, reporter molecules (4-Mercaptobenzoic acid, 4-MBA) embedded Au@Ag core-shell nanospheres as SERS nanotags were prepared for the detection of cTn I. As the Raman reporters were embedded between the core and the shell, they could be protected from the external environment and nanoparticle aggregation. Excellent SERS performances were obtained due to the enhanced local electromagnetic field in the gap of core and shell metals. In a standard phosphate buffered saline (PBS) environment, the limit of detection for cTn I was 0.0086 ng mL-1 (8.6 ppt) with a good linear relationship. The excellent Raman detection performance was attributed to the localized surface plasmon resonance effect and strong electromagnetic field enhancement effect produced by the gap between the Au core and the Ag shell. The SERS nanotags we prepared were facile to synthesize, and the analysis procedure could be completed quickly (15 min), which made the detection of cTn I faster. Therefore, the proposed SERS nanotags have significant potential to be a faster and more accurate tool for acute myocardial infarction diagnostics.

Rapid, Point-of-Care scFv-SERS Assay for Femtogram Level Detection of SARS-CoV-2.

Rapid, sensitive, on-site identification of SARS-CoV-2 infections is an important tool in the control and management of COVID-19. We have developed a surface-enhanced Raman scattering (SERS) immunoassay for highly sensitive detection of SARS-CoV-2. Single-chain Fv (scFv) recombinant antibody fragments that bind the SARS-CoV-2 spike protein were isolated by biopanning a human scFv library. ScFvs were conjugated to magnetic nanoparticles and SERS nanotags, followed by immunocomplex formation and detection of the SARS-CoV-2 spike protein with a limit of detection of 257 fg/mL in 30 min in viral transport medium. The assay also detected B.1.1.7 (“alpha”), B.1.351 (“beta”), and B.1.617.2 (“delta”) spike proteins, while no cross-reactivity was observed with the common human coronavirus HKU1 spike protein. Inactivated whole SARS-CoV-2 virus was detected at 4.1 × 104 genomes/mL, which was 10–100-fold lower than virus loads typical of infectious individuals. The assay exhibited higher sensitivity for SARS-CoV-2 than commercial lateral flow assays, was compatible with viral transport media and saliva, enabled rapid pivoting to detect new virus variants, and facilitated highly sensitive, point-of-care diagnosis of COVID-19 in clinical and public health settings.

An Electrochemical and Raman Scattering Dual Detection Biosensor for Rapid Screening and Biomolecular Profiling of Cancer Biomarkers

Detecting circulating biomarkers sensitively and quantitatively is paramount for cancer screening, diagnosis, and treatment selection. Particularly, screening of a panel of circulating protein biomarkers followed by mapping of individual biomarkers could assist better diagnosis and understanding of the cancer progression mechanisms. Herein, we present a miniaturized biosensing platform with dual readout schemes (electrochemical and Surface enhanced Raman scattering (SERS)) for rapid cancer screening and specific biomarker expressional profiling to support cancer management. Our approach utilizes a controlled nanomixing phenomena under alternative current electrohydrodynamic condition to improve the isolation of cancer-associated circulating proteins (i.e., Epidermal growth factor receptor (EGFR), BRAF, Programmed death-ligand 1 (PD-L1)) with antibody functionalized sensor surface for rapid and efficient isolation of the targets and subsequent labelling with SERS nanotags. The method employs Differential Pulse Voltammetry (DPV) for rapidly screening for the presence of the circulating proteins on biosensor surface irrespective of their type. Upon positive DPV detection, SERS is applied for sensitive read-out of individual biomarkers biomarker levels. In a proof-of-concept study, we demonstrate the dual detection biosensor for analysing circulating BRAF, EGFR and PDL-1 proteins and successfully screened both ensemble and individual biomarker expressional levels as low as 10 pg (1 ng/mL). Our findings clearly indicate the potential of the proposed method for cancer biomarker analysis which may drive the translation of this dual sensing concept in clinical settings.

SERS-based biosensing platform using shape-coded hydrogel microparticles incorporating silver nanoparticles

Biosensors based on surface-enhanced Raman scattering (SERS) represent an efficient tool that can quickly detect even very low concentrations of the target molecule. Recent SERS-based biosensing has been moving toward a multiplex detection platform to analyze samples containing a variety of molecules, such as blood and urine. In this study, shape-coded suspension arrays have been developed for SERS-based multiplex biosensing. The suspension microarrays were fabricated using different shapes of poly(ethylene glycol) (PEG)-based hydrogel microparticles, which were prepared by a simple photopatterning process. The resultant hydrogel particles were coated with silver nanoparticles by a silver mirror reaction and then divided into two groups depending on the shape. As a proof of concept, square-shaped particles were coated with 4-mercaptophenylboronic acid by self-assembly to detect glucose, while circular particles were functionalized with anti-human serum albumin (HSA) to detect HSA. The assay was performed in the concentration range of 1 pg/mL to 1 μg/mL for both glucose and HSA without crosstalk. Target selectivity was also confirmed from Raman mapping, which shows that each group of particles reveals a different pattern. Moreover, we verified the potential of our method in multiplex immunoassays by quantifying two different antigens using SERS nanotags. The performance of the developed SERS-based multiplex immunoassay platform was demonstrated by the detection of a very low concentration of biomolecules with excellent selectivity.

Simultaneous and accurate screening of multiple genetically modified organism (GMO) components in food on the same test line of SERS-integrated lateral flow strip

Herein, a surface-enhanced Raman scattering (SERS)-integrated LFS platform was developed for rapid and simultaneous screening of multiple genetically modified organism (GMO) components (promoter, codon, and terminator) in soybean. Research demonstrated that, on the same test line (T line) of single LFS, three different GMP components can be well distinguished with the help of three SERS nano tags. Good linear correlations between SERS signal and concentration of each GMO component were also obtained for quantitative analysis. Of greater importance, whether these multiple analytes coexisted or not, varied in the same concentration trend or not, these multiple GMP components can be rapidly (15 min) and accurately screened with satisfied sensitivity and specificity by decoding the signals on the same T line. We envision that this decoding platform can further improve the potential of LFS and SERS for practical applications and provide a promising alternative for multiple screening of GMO identification in food.

Simultaneous Detection of Inflammatory Biomarkers by SERS Nanotag-Based Lateral Flow Assay with Portable Cloud Raman Spectrometer.

Inflammatory biomarkers are closely related to infectious diseases. However, traditional clinical tests of laboratory inspection are unable to achieve rapid and accurate detection of these biomarkers on-site due to shortcomings such as complex experimental operation, expensive equipment, and long test time. Herein, we proposed a lateral flow assay (LFA) strip based on surface-enhanced Raman scattering (SERS) nanotags (SERS-LFA strips) for the simultaneous and quantitative detection of dual infection biomarkers, serum amyloid A (SAA) and C-reactive protein (CRP), respectively. In practice, mesoporous silica (mSiO2)-coated Au nanoparticles (Au NPs) were used as the SERS substrate. Mercaptobenzoic acid (MBA) was embedded in the internal gap between Au NPs and the mSiO2 shell to prepare AuMBA@mSiO2 NPs, onto which SAA and CRP antibodies were modified to prepare two AuMBA@mSiO2 SERS nanotags. The Raman intensities of the test and control lines were simultaneously identified for the qualitative detection of SAA and CRP, with limits of detection (LODs) as low as 0.1 and 0.05 ng/mL for SAA and CRP, respectively. Finally, aiming at point-of-care testing (POCT) applications, we used a smartphone-based portable Raman spectrometer to quantitatively analyze the SERS-LFA strips. The Raman signal could still be accurately detected when the concentration of SAA and CRP was 10 ng/mL, which is lower than the LOD required in clinical practice for most diseases. Therefore, taking into account its simple operation and short analysis time, by using a portable Raman spectrometer which can be equipped with a 5G cloud-based healthcare management system, the current strategy based on SERS-LFA provides the potential for the quick and on-site diagnosis of infectious diseases such as sepsis, which is of great significance for medical guidance on the treatment of widely spread infection-related diseases in remote areas that lack well-developed medical resources.

Surface-enhanced Raman scattering nanotags for bioimaging

Surface-enhanced Raman scattering (SERS) technique has shown extraordinary features for biomedical applications. The implementation of SERS nanotags has opened a new era for bioimaging and detections. As a powerful tool, SERS nanotags provide favorable properties such as fingerprint spectrum, narrow peak linewidth, good photostability, and high spatial resolution accompanied by various rational designs of nanoparticles. They have proven as useful imaging agents for in vivo, ex vivo, and in vitro detection of cancerous cells and tissues. This tutorial provides the basic principles of SERS and SERS nanotags, including recent progress of SERS-based bioimaging applications, as well as the outlooks into the future developments toward practical clinical SERS.

Aptamer-based novel Ag-coated magnetic recognition and SERS nanotags with interior nanogap biosensor for ultrasensitive detection of protein biomarker

• An innovative aptamer SERS biosensor for highly sensitive protein biomarker detection was presented. • The biosensor contains SERS tags and novel magnetic capture substrates. • Aptamers against CRP were modified on both SERS tag and magnetic capture substrate for specific recognition. • This method exhibits excellent selectivity and specificity and displays high accuracy in actual human serum samples. • This strategy can be extended to the detection of other biomarkers and hold great promise in the clinical diagnosis. The design of a rapid, simple, and ultrasensitive detection platform of protein biomarkers has become an urgent need in early diagnosis and treatment. Surface-enhanced Raman scattering (SERS) technology, with the unique advantages of ultra-sensitivity and fingerprint recognition, has attracted much attention in biomarker detection. An innovative aptamer SERS biosensor for highly sensitive protein biomarker detection was presented. This biosensor contains a SERS tag (reporter-labeled Au nano-bridged nanogaps particles, Au NNPs) and novel magnetic capture substrate (Ag-coated Fe 3 O 4 -Au NPs, Ag MNPs). C-reactive protein (CRP), a representative biomarker for diagnosing, was used to assess the SERS performance of this biosensor. Aptamers against CRP were modified on both SERS tag and magnetic capture substrate for specific recognition by Au NNPs-CRP-Ag MNPs sandwich assay. This strategy achieved sensitive detection of CRP with a limit of detection (LOD) of 10 fM (1.14 pg/mL), and is much lower than reported analytical methods. Besides, this method exhibits excellent selectivity and specificity for CRP under the interference of other proteins and displays high accuracy in the detection of actual human serum samples. This proposed detection strategy can be extended to the sensitive detection of other biomarker proteins and hold great promise in the clinical diagnosis of diseases.

Recent advances in applications of nanoparticles in SERS in vivo imaging.

Surface-enhanced Raman scattering (SERS) technique has been regarded as one of the most important research methods in the field of single-molecule science. Since the previous decade, the application of nanoparticles for in vivo SERS imaging becomes the focus of research. To enhance the performance of SERS imaging, researchers have developed several SERS nanotags such as gold nanostars, copper-based nanomaterials, semiconducting quantum dots, and so on. The development of Raman equipment is also necessary owing to the current limitations. This review describes the recent advances of SERS nanoparticles and their applications for in vivo imaging in detail. Specific examples highlighting the in vivo cancer imaging and treatment application of SERS nanoparticles. A perspective on the challenges and opportunities of nanoparticles in SERS in vivo imaging is also provided. This article is categorized under: Diagnostic Tools > in vivo Nanodiagnostics and Imaging Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.

Immunomagnetic Capture and Multiplexed Surface Marker Detection of Circulating Tumor Cells with Magnetic Multicolor Surface-Enhanced Raman Scattering Nanotags.

Circulating tumor cells (CTCs) have substantial clinical implications in cancer diagnosis and monitoring. Although significant progress has been made in developing technologies for CTC detection and counting, the ability to quantitatively detect multiple surface protein markers on individual tumor cells remains very limited. In this work, we report a multiplexed method that uses magnetic multicolor surface-enhanced Raman scattering (SERS) nanotags in conjunction with a chip-based immunomagnetic separation to quantitatively and simultaneously detect four surface protein markers on individual tumor cells in whole blood. Four-color SERS nanotags were prepared using magnetic-optical iron oxide-gold core-shell nanoparticles with different Raman reporters to recognize four different cancer markers with respective antibodies. A microfluidic device was fabricated to magnetically capture the nanoparticle-bound tumor cells and to perform online negative staining and single-cell optical detection. The level of each targeted protein was obtained by signal deconvolution of the mixed SERS signals from individual tumor cells using the classic least squares regression method. The method was tested with spiked tumor cells in human whole blood with three different breast cancer cell lines and compared with the results of purified cancer cells suspended in a phosphate buffer solution. The method, with either spiked cancer cells in blood or purified cancer cells, showed a strong correlation with purified cancer cells by enzyme-linked immunosorbent assay, suggesting the potential of our method for the reliable detection of multiple surface markers on CTCs. Combining immunomagnetic enrichment with high specificity, multiplexed targeting for the capture of CTC subpopulations, multicolor SERS detection with high sensitivity and specificity, microfluidics for handling rare cells and magnetic-plasmonic nanoparticles for dual enrichment and detection, our method provides an integrated, yet a simple and an efficient platform that has the potential to more sensitively detect and monitor cancer metastasis.

Fe3O4@Au SERS tags-based lateral flow assay for simultaneous detection of serum amyloid A and C-reactive protein in unprocessed blood sample

Accurate detection of mutiple biomarkers in unprocessed blood via lateral flow immunoassay (LFA) strips remains challenging. Herein, we reported a novel surface-enhanced Raman scattering (SERS)-based LFA strip for ultrasensitive and simultaneous analysis of dual infection biomarkers by using functionalized Fe3O4@Au magnetic nanoparticles (MNPs). Antibody-modified Fe3O4@Au MNPs were designed as SERS nanotags and efficient enrichment tool. The target infection biomarkers were quickly captured and enriched from the blood sample by Fe3O4@Au nanotags. After washing to remove impurities, Fe3O4@Au nanotags acted as high-performance SERS reporters for quantitative detection of the target in the LFA strip. Through this strategy, the proposed Fe3O4@Au based LFA allowed quantified analysis of target infection biomarkers in unprocessed blood samples with the limits of detection of 0.1 ng/mL and 0.01 ng/mL for serum amyloid A (SAA) and C-reactive protein (CRP), respectively. By comparison, the proposed assay for SAA and CRP was 100-fold and 1000-fold more sensitive than standard colloidal gold strip by using the same antibodies, respectively. This magnetic SERS strip also exhibited excellent stability, specificity, and selectivity for complex samples and could be used as an effective tool for infectious disease diagnostics.

SERS biosensors for ultrasensitive detection of multiple biomarkers expressed in cancer cells

The design and fabrication of multifunctional surface-enhanced Raman scattering (SERS) nanotags are key issues in their application to biological imaging of cells and tissues. In this study, highly sensitive, reproducible and long-term stable SERS nanotags were developed for the identification of localized distribution of multiple protein biomarkers expressed on breast cancer cells. To enhance the surface electromagnetic fields of Raman reporter molecules, Ag-encapsulated Au (Ag-Au) hollow nanospheres were synthesized. Strong Raman signal enhancement effects could be achieved by positioning Raman reporter molecules in nanogaps between the Au hollow nanospheres and silver shell. In addition, the signal was also enhanced due to the localization of surface electromagnetic fields through the pinholes on the surface of Au hollow nanospheres. To maintain the long-term stability of the Au hollow-Ag core/shell nanospheres, their surface was coated with a polyethylene glycol (PEG) layer. The biocompatibility of PEGylated Ag-Au hollow nanospheres was investigated using the premix water soluble tetrazolium salt (WST-1) cell viability test. These SERS nanotags also enabled a high-resolution multiplexed live cell imaging. Our proposed SERS imaging technique using the new SERS nanotags provides a new platform for fast and accurate classification of different phenotypes of breast cancer cells.

A boronate-modified molecularly imprinted polymer labeled with a SERS-tag for use in an antibody-free immunoassay for thecarcinoembryonic antigen.

An antibody-free immunoassay that makes use of a boronate affinity molecularly imprinted polymer (MIP) and surface enhanced Raman scattering (SERS) tags is described. It was applied to the specific determination of the carcinoembryonic antigen (CEA) in human serum. For the preparation of the boronate affinity array, a polymer capable of adsorbing glycoproteins was first synthesized on the surface of a glass slide with four spots using 4-vinylbenzeneboronic acid (VPBA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the crosslinking agent, and ethylene glycol and cyclohexanol as porogens. The surface of the VPBA-Co-EGDMA can bind target glycoproteins. After specific capture of the glycoprotein, a "MIP-target glycoprotein-SERS tag" sandwich structure was formed by covalent interaction between the SERS nanotag (consisting of gold nanoparticles and 4-mercaptophenylboronic acid [MPBA]). CEA can be quantified in spiked serum with a detection limit of 0.1ng·mL-1 via the specific Raman band at 1098cm-1. Graphical abstractSchematic representation of the boronate affinity molecularly imprinted polymer (MIPs) array-based SERS sensor for rapid and sensitive detection of thecarcinoembryonic antigen (CEA) from human serum. The boronate affinity MIPs array are used as capture probes, and MPBA@AuNPs are used as SERS tags.

SERS-Based Immunoassays for the Detection of Botulinum Toxins A and B Using Magnetic Beads.

Rapid and sensitive detection of botulinum neurotoxins (BoNTs) is important for immediate treatment with proper antitoxins. However, it is difficult to detect BoNTs at the acute phase of infection, owing to its rarity and ambiguous symptoms. To resolve this problem, we developed a surface-enhanced Raman scattering (SERS)-based immunoassay technique for the rapid and sensitive detection of BoNTs. Magnetic beads and SERS nanotags as capture substrates and detection probes, respectively, and Nile Blue A (NBA) and malachite green isothiocyanate (MGITC) as Raman reporter molecules were used for the detection of two different types of BoNTs (types A and B), respectively. The corresponding limits of detection (LODs) were determined as 5.7 ng/mL (type A) and 1.3 ng/mL (type B). Total assay time, including that for immunoreaction, washing, and detection, was less than 2 h.

A Molecular Beacon Based Surface-Enhanced Raman Scattering Nanotag for Noninvasive Diagnosis of Bladder Cancer.

Current techniques responsible for bladder cancer diagnosis and monitoring are insensitive and invasive. Here, we report a surface-enhanced Raman scattering nanotag for the sensitive diagnosis of bladder cancer using urine samples as a noninvasive approach. The sea-urchin-like Au nanoclusters used in this work exhibit excellent surface-enhanced Raman scattering ability with an enhancement factor of 3.44 × 107. Molecular beacons labeled with Cy3 are covalently anchored to the surface of Au nanoclusters, which serve as a specific recognition site for survivin mRNA. Further a polyethylene glycol coating provides stability and completes the final functionalization. This surface-enhanced Raman scattering nanotag has good efficiency (equilibrium time: 10 min) with high sensitivity (detection threshold: 19.4 nM), high specificity (capable of single-base mismatch recognition) and good stability against nucleases. All these features are also verified in the fluorescence modality. Furthermore, its function was highly maintained in clinical samples from 13 patients with bladder cancer, as evidenced by a sensitivity up to 91.7% and a specificity up to 100%. The nanotag demonstrates its superiority over cytology and has its great clinical value even for early bladder cancer diagnosis. Thus, the nanotag is promising for noninvasive and sensitive diagnosis of bladder cancer.

SERS-Based Pump-Free Microfluidic Chip for Highly Sensitive Immunoassay of Prostate-Specific Antigen Biomarkers.

Highly sensitive analysis of cancer biomarkers demonstrates an important impact in early diagnosis and therapies of cancer. A novel surface-enhanced Raman scattering (SERS) based immunoassay using microfluidic technique was reported for rapid analysis of prostate-specific antigen (PSA) biomarker. It is a useful screening test to discriminate prostate cancer and other diseases related to prostate. A "sandwich" immunoassay based on SERS nanotags, PSA biomarkers, and magnetic beads was applied on a pump-free microfluidic sensor. Magnetic immunocomplexes are isolated and trapped at the detection chamber by a permanent magnet integrated into the chip. The PBS buffer washed magnetic immunocomplexes and brought the free gold nanoparticles to the downsteam channel for waste. Our results show a good linear response in the range from 0.01 to 100 ng mL-1. The limit of detection of the PSA level is estimated to be below 0.01 ng mL-1 using this chip. This detection level of PSA biomarker in human serum can be accomplished in 5 min without manual incubation and a heavy syringe pump. To the best of our knowledge, this is the first SERS-based immunoassay which applied a pump-free microfluidic chip as a detection platform. We believe that the proposed method reveals a valuable potential tool for the diagnosis of prostate cancer.

A high-resolution study of in situ surface-enhanced Raman scattering nanotag behavior in biological systems

The colloidal stability of surface-enhanced Raman scattering (SERS) nanotags (Raman reporter-conjugated plasmonic nanoparticles) significantly affects the accuracy and reproducibility of SERS measurements, particularly in biological systems. Limited understanding of SERS nanotag stability may partly hamper the translation of SERS nanotags from the laboratory to their use in the clinic. In this contribution, we utilized differential centrifugal sedimentation (DCS), a reliable and straightforward technique to comprehensively analyze the colloidal stability of SERS nanotags in biological systems. Compared with other particle characterization techniques, DCS has been shown to have a unique advantage for high-resolution and high-throughput polydisperse particle characterization. DCS data revealed that the universal aggregation prevention practice of coating SERS nanotags with silica or bovine serum albumin layers did not sufficiently stabilize them in common measurement environments (e.g., 1 × PBS). Combined DCS and SERS measurements established a strong correlation between the degrees of nanotag aggregation and signal intensities, further reinforcing the necessity of characterizing SERS nanotag stability for every condition in which they are used. We also found that increasing the protein thickness by the inclusion of extra protein components in the detection environments and antibody functionalization can improve the stability of SERS nanotags. We believe that this study can provide guidelines on appropriate measurement techniques and particle design considerations to assess and improve SERS nanotag stability in complex biological systems.