Reproducible Surface-enhanced Raman Scattering Spectra Research Articles

Impact of silver nanoparticles size on SERS for detection and identification of filamentous fungi

Using the proper size of nanoparticles as an active substrate, Surface-enhanced Raman scattering (SERS) can provide a reliable technique for detecting and identifying fungi, including Alternaria alternata, Aspergillus flavus, Fusarium verticilliodes, and Aspergillus parasiticus that have been associated to biodeterioration and biodegradation of cultural heritage materials. In this research spherical silver nanoparticles (AgNPs) of average size of 10, 30 and 60 nm were synthesized using the wet chemical method with good yield and their size and shape distributions were examined using small-angle X-ray scattering (SAXS). The protocol for fungi sample preparation proved to be critical for producing high-quality and reproducible SERS spectra. We found that the effect of AgNPs on SERS signal enhancement is size dependent under the same experimental conditions; the SERS intensity of fungal strains using 60 nm achieved up to 2.3x105 enhancement, about twice as intense as those produced with 30 nm, and 10 nm produced a minor broad weak peak barely discernible around 1400 cm−1, similar to the NR spectra profile in the 550–1700 cm−1 spectral region, and the SERS signals using 60 nm showed high reproducibility, with less than 20% variance. Furthermore, we used principal component analysis (PCA) to statistically classify the SERS spectrum into four separate clusters with 99 percent variability so that the four fungal strains could be clearly detected and identified. The SERS technique, in combination with the PCA developed in this study, provides a simple, rapid, accurate, and cost-effective analytical tool for detecting and identifying filamentous fungal strains.

Improvement of Surface-Enhanced Raman Scattering Method for Single Bacterial Cell Analysis.

Surface-enhanced Raman scattering (SERS) is a useful tool for label-free analysis of bacteria at the single cell level. However, low reproducibility limits the use of SERS. In this study, for the sake of sensitive and reproducible Raman spectra, we optimized the methods for preparing silver nanoparticles (AgNPs) and depositing AgNPs onto a cell surface. We found that fast dropwise addition of AgNO3 into the reductant produced smaller and more stable AgNPs, with an average diameter of 45 ± 4 nm. Compared with that observed after simply mixing the bacterial cells with AgNPs, the SERS signal was significantly improved after centrifugation. To optimize the SERS enhancement method, the centrifugal force, method for preparing AgNPs, concentration of AgNPs, ionic strength of the solution used to suspend the cells, and density of the cells were chosen as impact factors and optimized through orthogonal experiments. Finally, the improved method could generate sensitive and reproducible SERS spectra from single Escherichia coli cells, and the SERS signals primarily arose from the cell envelope. We further verified that this optimal method was feasible for the detection of low to 25% incorporation of 13C isotopes by the cells and the discrimination of different bacterial species. Our work provides an improved method for generating sensitive and reproducible SERS spectra.

Surface plasmon resonance and surface-enhanced Raman scattering activity of SiO2–Au core-cap nanostructure arrays

SiO2–Au core-cap nanostructure arrays were prepared by dip-coating technique combined with wet chemical reduction method. The surface morphologies, structures, and optical properties of the obtained samples were characterized by scanning electron microscopy, X-ray diffraction, and ultraviolet–visible spectrophotometer, respectively. The surface-enhanced Raman scattering (SERS) activity of SiO2–Au core-cap nanostructure arrays substrates was investigated using leucine as probe molecule. And the relationship between the SERS effect and the surface plasmon resonance (SPR) peaks was discussed. High-quality, stable, and reproducible SERS spectra of leucine were successfully obtained. When the maximum SPR peak matched with the excitation wavelength, the substrate gave rise to the highest SERS enhancement. Furthermore, six different fluorescent dyes were also chosen as probe molecules. It was found that the substrate showed good Raman enhancement and highly efficient fluorescence quenching characteristic on these fluorescent dyes.

Paper-based plasmonic platform for sensitive, noninvasive, and rapid cancer screening

Surface-enhanced Raman scattering (SERS) fingerprints of individual molecules offer the possibility of multiplexing as well as cancer screening. A highly sensitive, noninvasive, and rapid cancer screening platform encompassing exfoliative cytology and paper-based SERS technology is described. The SERS substrate which consists of plasmonic gold nanorods (GNRs) adsorbed on a piece of filter paper forms the flexible and three-dimensional heterogeneous scaffold for cancer screening. Different and reproducible SERS spectra are obtained from normal and cancerous cells due to specific biomolecular changes in cancerous cells. A diagnostic algorithm based on the ratio of the spectra values is adopted to distinguish between cells exfoliated from 20 normal and cancerous tissues, and a high sensitivity of 100% and specificity of 100% are achieved by I1600/1440 (peak ratio of signals at 1600–1440cm−1) and I1440/1340 (1440–1340cm−1), which is better than I1600/1340 (1600–1340cm−1) with a sensitivity of 70% and specificity of 60%. The combination of exfoliative cytology and paper-based plasmonic technology enables highly sensitive, rapid, and non-invasive cancer screening and has large clinical potential.

One-step fabrication of sub-10-nm plasmonic nanogaps for reliable SERS sensing of microorganisms

Nanoscale gaps in noble metal films can produce intense electromagnetic enhancement. When Raman-active molecules are positioned in these regions, their surface-enhanced Raman scattering (SERS) signals can be dramatically enhanced. However, the lack of convenient and reliable fabrication methods with ultrasmall nanogaps (<10nm) severely block the application of SERS. Here, we propose a cost-effective and reproducible technique to fabricate the large-area Ag SERS-active substrates which are full of the high-density, sub-10-nm nanogaps by high pressure sputtering, and the enhancement factor (EF) is testified to improve by 103 times compared to the continuous Ag film with a smooth surface (the roughness is 0.5nm) and without nanogaps. Since there are no chemicals used during fabrication, this substrate has a clean surface, which is crucial for acquiring reliable SERS spectra. This SERS-active substrate has then been applied to identify a series of microorganisms, and excellent, reproducible SERS spectra were obtained. Finally, a set of piecewise-linear equations is provided according to the correlation between SERS intensity and rhodamine 6G (R6G) concentration, and the detection limit is calculated to be 0.2×10−8M. These results suggest that the high pressure sputtering is an excellent, reliable technique for fabricating sub-10-nm plasmonic nanogaps, and the SERS-based methodology is very promising for being used in biological sensing field.

Proline and hydroxyproline deposited on silver nanoparticles. A Raman, SERS and theoretical study

The Raman and surface‐enhanced Raman scattering (SERS) spectra of l‐proline (Pro) and trans‐4‐hydroxy‐ l‐proline (Hyp) were recorded. SERS spectra were obtained on colloidal Ag prepared by reduction with hydroxylamine. Allowing sufficient time for Pro and Hyp to adjust in the colloidal solution resulted fundamentally in obtaining unique and reproducible SERS spectra. Hyp stabilizes on the surface more rapidly than Pro. The spectral analysis indicates that Pro interacts with the Ag surface through the carboxylate group. The interaction of Hyp with the metal surface occurs through the amino, methylene and carboxylate moieties of the molecule. The spectroscopic results are supported by quantum chemical calculations, performed using extended Hückel theory (EHT) of the title compounds interacting with an Ag cluster model. The assignment of the Raman bands was supported by a normal coordinate analysis performed through Becke, three‐parameter, Lee–Yang–Parr/6‐311 G* + calculations. Copyright © 2011 John Wiley &amp; Sons, Ltd.

Multiplexing with SERS labels using mixed SAMs of Raman reporter molecules

Surface-enhanced Raman scattering (SERS) offers a tremendous multiplexing capacity for the selective detection of biomolecules in targeted research. SERS labels comprising self-assembled monolayers (SAMs) of Raman reporter molecules on the surface of metal nanoparticles are sensitive and robust probes. Advantages of a SAM include maximum sensitivity, minimal unwanted co-adsorption of molecules from the surroundings, and reproducible SERS spectra with only few dominant Raman bands--all of these independent of a particular SERS substrate. We demonstrate experimentally how to increase the multiplexing capacity of SERS labels by using mixed SAMs with up to three different Raman reporter molecules on the surface of the metal colloid. Type and stoichiometry of a particular Raman label in a multi-component SAM are additional parameters compared with one-component SAMs. All one-, two-, and three-component SAMs on gold nanospheres can be easily discriminated, either by their original SERS spectra or the corresponding bar codes.

In Situ Surface-Enhanced Raman Scattering Analysis of Biofilm

Biofilms represent the predominant form of microbial life on Earth. They are aggregates of microorganisms embedded in a matrix formed by extracellular polymeric substances (EPS). Detailed information about chemical composition and structure of the EPS matrix is relevant e.g. for the optimization of biocides, of antifouling strategies and for biological wastewater treatment. Raman microscopy (RM) is a capable tool that can provide detailed chemical information about biofilm constituents with spatial resolution of optical microscope. However, the sensitivity of RM is limited. Surface-enhanced Raman scattering (SERS), which enables investigations of biomolecules at very low concentration levels, allows overcoming this drawback. To our knowledge, this paper is the first report on reproducible SERS spectra from different constituents of a multispecies biofilm. We believe that the reproducibility is partly owed to the in situ measurement of the biofilm, while up to now SERS measurements of microbiological samples by RM were carried out after sample drying. We employed colloidal silver nanoparticles for in situ SERS measurements by RM. The achieved enhancement factor of up to 2 orders of magnitude illustrates a high potential of SERS for ultrasensitive chemical analysis of biofilms, including the detection of different components and the determination of their relative abundance in the complex biofilm matrix.

Towards single-microorganism detection using surface-enhanced Raman spectroscopy

The identification and discrimination of microorganisms is important not only for clinical reasons but also for pharmaceutical clean room production and food-processing technology. Vibrational spectroscopy such as IR, Raman, and surface-enhanced Raman scattering (SERS) can provide a rapid ‘fingerprint’ on the chemical structure of molecules and is used to obtain a ‘fingerprint’ from microorganisms as well. Because of the requirement that a single bacterium cell and noble metal nanoparticles must be in close contact and the lack of a significant physical support to hold nanoparticles around the single bacterium cell, the acquisition of SERS spectra for a single bacterium using colloidal nanoparticles could be a challenging task. The feasibility of SERS for identification down to a single bacterium is investigated. A Gram-negative bacterium, Escherichia coli, is chosen as a model for the investigation. Because the adsorption of silver nanoparticles onto the bacterial cell is an exclusive way for locating nanoparticles close to the bacterium cell, the absorption characteristics of silver nanoparticles with different surface charges are investigated. It is demonstrated that the citrate-reduced colloidal silver solution generates more reproducible SERS spectra. It is found that E. coli cells aggregate upon mixing with silver colloidal solution, and this may provide an additional benefit in locating the bacterial cell under a light microscope. It is also found that a laser wavelength in the UV region could be a better choice for the study due to the shallow penetration depth. It is finally shown that it is possible to obtain SERS spectra from a single cell down to a few bacterial cells, depending on the aggregation properties of bacterial cells for identification and discrimination.

Reproducible Surface-Enhanced Raman Scattering Spectra of Bacteria on Aggregated Silver Nanoparticles

Surface-enhanced Raman scattering (SERS) is proven to be a powerful technique for rapid identification and discrimination of microorganisms. However, due to the heterogeneous nature of the samples, the acquisition of reproducible spectra hinders the further development of the technique. In this study, we demonstrate the influence of the experimental conditions on SERS spectra. Then, we report a simple sample preparation method coupled with a light microscope attached to a Raman spectrometer to find a proper spot on the sample to acquire reproducible SERS spectra. This method utilizes the excited surface plasmons of the aggregated silver nanoparticles to visualize the spots on the sample. The samples are prepared using the concentrated silver colloidal solutions. The collection time for one spectrum is 10 s and each spectrum is a very good representative of the other spectra acquired from the same sample. The nature of the surface charge of the silver nanoparticles influences the spectral features by determining the strength of the interactions between nanoparticles and bacteria and the aggregation properties of the nanoparticles. Although increasing the colloid concentration in the sample resulted in reproducible spectra from arbitrary points on the sample, a great variation from sample to sample prepared with the different colloidal solution concentrations is observed.