Label-free detection of proteins from dried-suspended droplets using surface enhanced Raman scattering

A simple sample preparation method to obtain rich and reproducible surface-enhanced Raman scattering (SERS) spectra from proteins regardless of their surface properties and dimensions for label-free detection and identification is reported. The method uses colloidal silver nanoparticles (AgNPs) as substrates and is based on suspending the droplet of a mixture containing AgNPs and proteins from a hydrophobic surface. Drying the droplet at this suspended configuration allows the accumulation and packing of AgNPs and protein molecules in the middle of the droplet area rather than getting jammed at the edges of drying droplets as the solvent evaporates. A detection limit of down to 0.05 μg mL(-1) for some of the model proteins used in this study is obtained with this simple approach. The advantage of this method is its simplicity and improved sensitivity over other approaches reported in the literature.     

Determining the composition of aqueous microdroplets with broad-band cavity enhanced Raman scattering

We demonstrate that broad-band cavity enhanced Raman scattering (CERS) can be used to determine the composition of binary alcohol-water aerosol droplets over a wide compositional range from 10% v/v to 90% v/v. In contrast to conventional CERS using narrow-band laser excitation, the excitation is provided by a broad-band Nd:YAG pumped dye laser. A change in the spontaneous spectrum resulting from the change of the linewidth of the excitation laser permits tuning of the sensitivity range over which the droplet composition can be determined by CERS. We demonstrate that this change in sensitivity can be estimated using a simulation of the change in the sensitivity to the species in spontaneous bulk phase measurements. We further show that the compositional calibration is independent of droplet radius in the range 33-56 microm. The compositional range over which CERS is sensitive can be controlled and optimised for any particular application by exploiting the dependence of the stimulated Raman scattering on the laser linewidth and wavelength. Thus, quantitative measurements of droplet composition can be made in situ with high accuracy, providing a valuable new tool for analysing aerosol composition.     

Probing the Raman scattering tensors of individual molecules

Single-molecule experiments provide new views into the mechanisms behind surface-enhanced Raman scattering. It was shown previously that spectra of individual rhodamine 6G molecules adsorbed on silver nanocrystal aggregates present stronger fluctuations in two low-frequency bending modes, at 614 and 773 cm(-1). Here we use polarization spectroscopy to show that these bands are enhanced by a resonant process whose transition dipole is rotated by 15+/-10 degrees with respect to the molecular transition dipole. We also show that the polarization function remains stable over the whole time scale of a measurement, indicating that molecular reorientation with respect to the surface is unlikely. Together these findings provide further support to the involvement of a charge-transfer resonance in the enhancement of the low-frequency bands and allow us to suggest a model for the orientation of rhodamine 6G molecules at Raman hot spots.     

Surface enhanced resonance Raman scattering detection by fluorimeter

A commercially available fluorimeter with a white light source is used to detect surface enhanced resonance Raman scattering (SERRS). This approach allows facile tunability of the excitation source for SERRS.     

Raman thermometry measurements of free evaporation from liquid water droplets

Recent theoretical and experimental studies of evaporation have suggested that on average, molecules in the higher-energy tail of the Boltzmann distribution are more readily transferred into the vapor during evaporation. To test these conclusions, the evaporative cooling rates of a droplet train of liquid water injected into vacuum have been studied via Raman thermometry. The resulting cooling rates are fit to an evaporative cooling model based on Knudsen's maximum rate of evaporation, in which we explicitly account for surface cooling. We have determined that the value of the evaporation coefficient (gamma(e)) of liquid water is 0.62 +/- 0.09, confirming that a rate-limiting barrier impedes the evaporation rate. Such insight will facilitate the formulation of a microscopic mechanism for the evaporation of liquid water.     

Surface enhanced Raman scattering of ammonia

A surface enhanced Raman scattering (SERS) spectrum of 0.5 M NH₃ in 4.0 M KCl has been observed on a silver electrode. An approximate enhancement factor of 3 × 10⁵ is calculated, and additional evidence for the enhanced nature of the spectrum is provided by the observation that totally symmetric vibrations are depolarized and by the strong potential dependence of the intensity of surface lines. Assignments have been given to the SERS lines with the low-frequency lines assigned to a AgCl and AgN stretch. The positive shift of the maximum of the intensity versus voltage curve with a lower laser excitation frequency is taken as evidence for the occurrence of a charge transfer process from ammonia to the silver electrode. The fact that the SERS spectrum of NH₃ on Ag can only be observed at large electrolyte concentrations is attributed to the breaking of hydrogen bonding at the electrode-solution interface.     

Probing the structure of single-molecule surface-enhanced Raman scattering hot spots

We present here a detailed study of the specific nanoparticle structures that give rise to single-molecule surface-enhanced Raman scattering (SMSERS). A variety of structures are observed, but the simplest are dimers of Ag nanocrystals. We chose one of these structures for detailed study using electrodynamics calculations and found that the electromagnetic SERS enhancement factors of 10(9) are easily obtained and are consistent with single-molecule SERS activity.     

Probing nanoscale interfaces with electrochemical surface-enhanced Raman scattering

Surface-enhanced Raman scattering (SERS) is a powerful tool for studying nanoscale molecule-metal interfaces across a range of electrochemical applications. SERS combines molecular-level information with a high degree of surface specificity, making it an ideal tool for understanding interfacial processes, from understanding how analytes and electrolytes organize near metal surfaces to following surface-mediated reactions in real time. However, because SERS relies on the excitation of localized surface plasmons, additional effects such as the production of hot charge carriers and photothermal heating can impact electrochemical SERS signals. These effects must also be considered when using SERS for quantitative electrochemical studies.     

Surface enhanced resonance Raman scattering of the perylene chromophore

Vibrational fundamentals, overtones and combination bands of the perylene chromophore, in the N-hexyl-3, 4:9,10-perylenetetracarboxylic diimide (HPTCNH) and other perylene tetracarboxylic derivatives, have been observed using surface enhanced resonance Raman scattering (SERRS) of Langmuir—Blodgett (LB) monolayers on Ag island films. Typical vibrational progressions due to the Franck—Condon (A-term) were seen. The results showed that the mechanism of the RRS effect was not altered by the metal surface, although the RRS signal was enhanced by four orders of magnitude. Polarization properties of the SERRS signal were studied for LB monolayers on a series of SERS active substrates. A frequency dependence of the depolarization ratios was observed.     

Surface enhanced Raman scattering for multiplexed detection

The multiplexed detection of biological analytes from complex mixtures is of crucial importance for the future of intelligent management and detection of disease. This review focuses on recent advances in the use of surface enhanced Raman scattering (SERS) spectroscopy as an analytical technique that can deliver multiplexed detection for a variety of biological target in increasingly complex media. The use of SERS has developed from the multipelxed detection of custom dye molecules to biomolecules such as DNA and proteins. Recent work has also shown the capability of SERS multiplexing for in vivo as well as in vitro applications.     

Practical understanding and use of surface enhanced Raman scattering/surface enhanced resonance Raman scattering in chemical and biological analysis

The unique ability to obtain molecular recognition of an analyte at very low concentrations in situ in aqueous environments using surface enhanced Raman scattering (SERS) and surface enhanced resonance Raman scattering (SERRS) detection makes these spectroscopies of considerable interest. Improved understanding of the effect coupled to improvements in practical techniques make the use of SERS/SERRS much simpler than has been the case in the past. This article is designed as a tutorial review targeted at aiding in the development of practical applications.     

DNA detection by surface enhanced resonance Raman scattering (SERRS)

This Education article outlines the different ways in which surface enhanced resonance Raman scattering (SERRS) can be used for the detection of DNA. The use of various different SERRS detection strategies that have allowed both sensitive and selective detection to be obtained is covered. Detection of DNA by SERRS involves the use of a dye with the DNA, whether as an intercalator or by direct covalent attachment. This generates strong SERRS signals that indicate the presence of the specific DNA sequence. The SERRS detection of DNA in different molecular biological assays is also discussed.     

Determination of the size and composition of multicomponent ethanol/water droplets by cavity-enhanced Raman scattering

Cavity-enhanced Raman scattering is used to determine the size and composition of multicomponent ethanol/water droplets in the concentration range 7.5–19% ethanol by volume. Under the experimental conditions presented here, the integrated CERS signal from ethanol shows an exponential increase with increase in ethanol concentration when compared with the integrated intensity of the water band. The calibration is shown to be invariant with particle size over the droplet radius range 20–35 μm. In addition to providing a method for determining particle size and composition, initial studies show that the evaporation dynamics of these multicomponent droplets can be probed by CERS.     

The role of cavity sites in surface-enhanced Raman scattering

Thermal desorption spectra (TDS) of pyridine from silver films deposited in ultra high vacuum are reported. Marked differences in the TDS are seen depending on the deposition conditions and the thermal history of the films, which have been correlated with surface-enhanced Raman scattering (SERS). These results as well as some of the observations in electrochemical systems are discussed in light of the recent Xe probe analysis carried out by Albano et al.     

Single molecule level detection of allophycocyanin by surface enhanced resonance Raman scattering

Single molecule level detection of the near-infrared fluorescent protein allophycocyanin (APC) has been achieved using surface enhanced resonance Raman scattering (SERRS). The detection limit using the peak height of the 440 cm(-1) band was 1 x 10(-13) mol l(-1), compared to 2 x 10(-12) mol l(-1) for the fluorescence peak at 660 nm.     

Gold particle interaction in regular arrays probed by surface enhanced Raman scattering

Lithographically designed two-dimensional arrays consisting of gold nanoparticles deposited on a smooth gold film are used as substrate to examine the SERS effect of the trans-1,2-bis (4-pyridyl) ethylene molecule. These arrays display two plasmon bands instead of the single one observed for the same arrays of particles but deposited on indium tin oxide coated glass. Laser excitation within the short wavelength band does not bring about any SERS spectrum, while excitation within the long wavelength band yields SERS spectra with a gain per molecule rising up to 10(8). The simultaneous investigation of extinction and Raman spectra of arrays exhibiting various topography parameters enables us to suggest an interpretation for both the occurrence of the two plasmon resonances and for the high Raman enhancement. We suggest to assign the short wavelength band to a plasmon wave propagating at the gold glass interface and the long wavelength one to an air/gold surface plasmon mode modified by particle-particle interaction.     

Surface enhanced Raman scattering arising from multipolar plasmon excitation

Visible and near infrared extinction spectra of gold nanorod regular arrays exhibit several bands assigned to high multipolar order plasmon resonances. These up to ninth order multipolar resonances generate surface enhanced Raman scattering spectra with typically 5 x 10(4) enhancement which is of similar magnitude as those obtained for dipolar excitations.     

Semi-quantitative trace analysis of nuclear fast red by surface enhanced resonance Raman scattering

The dye nuclear fast red has been detected and determined semi-quantitatively by means of surface enhanced resonance Raman scattering (SERRS) and surface enhanced Raman scattering (SERS), using laser exciting wavelengths of 514.5 and 632.8 nm, respectively, by employing a citrate-reduced silver colloid. A good linear correlation is observed for the dependence of the intensities of the SERRS bands at 989 cm⁻¹ (R=0.9897) and 1278 cm⁻¹ (R=0.9872) on dye concentration over the range 10⁻⁹ to 10⁻⁷ M, when using an exciting wavelength of 514.5 nm. At dye concentrations above 10⁻⁷ M, the concentration dependence of the SERRS signals is non-linear. This is almost certainly due to the coverage of the colloidal silver particles being in excess of a full monolayer of the dye. A linear correlation is also observed for the dependence of the intensities of the SERS bands at 989 cm⁻¹ (R=0.9739) and 1278 cm⁻¹ (R=0.9838) on the dye concentration over the range 10⁻⁸ to 10⁻⁶ M when using an exciting wavelength of 632.8 nm. Strong fluorescence prevented collection of resonance Raman scattering (RRS) spectra from powdered samples or aqueous solutions of the dye using an exciting wavelength of 514.5 nm, but weak bands were observed in the spectra obtained from both powdered and aqueous samples of the dye using an exciting wavelength of 632.8 nm. A study of the pH dependence of SERRS/SERS and UV–VIS absorption spectra revealed the presence of different ionisation states of the dye. The limits of detection for nuclear fast red by SERRS (514.5 nm), SERS (632.8 nm) and visible spectroscopy (535 nm) are 9, 89 and 1000 ng ml⁻¹, respectively.     

Metal-molecule Schottky junction effects in surface enhanced Raman scattering

We propose a complementary interpretation of the mechanism responsible for the strong enhancement observed in surface enhanced raman scattering (SERS). The effect of a strong static local electric field due to the Schottky barrier at the metal-molecule junction on SERS is systematically investigated. The study provides a viable explanation to the low repeatability of SERS experiments as well as the Raman peak shifts as observed in SERS and raw Raman spectra. It was found that a strong electrostatic built-in field at the metal-molecule junction along specific orientations can result in 2-4 more orders of enhancement in SERS.     

Surface enhanced Raman scattering based on silver dendrites substrate

A simple method of the reduction of AgNO3 by copper foil in aqueous medium was used to prepare silver dendrites, which can be used as a novel good reproducible surface enhanced Raman scattering (SERS) active substrate. The SERS spectra of 4-pyridinethiol on this novel substrate reflected the different SERS activities on the minuteness and strong Ag dendrites. The electromagnetic coupling enhancement and chemical enhancement mechanisms are used to explain the SERS effect.