Surface-enhanced Raman spectroscopy (SERS) combines molecular fingerprint specificity with potential single-molecule sensitivity. Therefore, the SERS technique is an attractive tool for sensing molecules in trace amounts within the field of chemical and biochemical analytics. Since SERS is an ongoing topic, which can be illustrated by the increased annual number of publications within the last few years, this review reflects the progress and trends in SERS research in approximately the last three years. The main reason why the SERS technique has not been established as a routine analytic technique, despite its high specificity and sensitivity, is due to the low reproducibility of the SERS signal. Thus, this review is dominated by the discussion of the various concepts for generating powerful, reproducible, SERS-active surfaces. Furthermore, the limit of sensitivity in SERS is introduced in the context of single-molecule spectroscopy and the calculation of the 'real' enhancement factor. In order to shed more light onto the underlying molecular processes of SERS, the theoretical description of SERS spectra is also a growing research field and will be summarized here. In addition, the recording of SERS spectra is affected by a number of parameters, such as laser power, integration time, and analyte concentration. To benefit from synergies, SERS is combined with other methods, such as scanning probe microscopy and microfluidics, which illustrates the broad applications of this powerful technique.

Raman spectroscopy is a valuable tool in various research fields. The technique yields structural information from all kind of samples often without the need for extensive sample preparation. Since the Raman signals are inherently weak and therefore do not allow one to investigate substances in low concentrations, one possible approach is surface-enhanced (resonance) Raman spectroscopy. Here, rough coin metal surfaces enhance the Raman signal by a factor of 10(4)-10(15), depending on the applied method. In this review we discuss recent developments in SERS spectroscopy and their impact on different research fields.

The application of a liquid/liquid microsegmented flow for serial high-throughput microanalytical systems shows promising prospects for applications in clinical chemistry, pharmaceutical research, process diagnostics, and analytical chemistry. Microscopy and microspectral analytics offer powerful approaches for the analytical readout of droplet based assays. Within the generated segments, individuality and integrity are retained during the complete diagnostic process making the approach favored for analysis of individual microscaled objects like cells and microorganisms embedded in droplets. Here we report on the online application of surface-enhanced micro-Raman spectroscopy for the detection and quantization of analytes in a liquid/liquid segmented microfluidic system. Data acquisition was performed in microsegments down to a volume of 180 nl. With this approach, we overcome the well-known problem of adhesion of colloid/analyte conjugates to the optical windows of detection cuvettes, which causes the so-called "memory effect". The combination of the segmented microfluidic system with the highly sensitive SERS technique reaches in a reproducible quantification of analytes with the SERS technique.

The development of fast identification techniques of viruses is an ongoing important research topic. Conventional virus detection and identification is generally based on various different microbiological methods. However, these techniques are not suitable for the analysis of single virus particles. Therefore, our goal is to establish tip-enhanced Raman scattering (TERS), providing vibrational spectroscopic information with a spatial resolution less than 50 nm, to characterize single viruses at a molecular level. Here we report, to the best of our knowledge for the first time, about TERS spectra of a tobacco mosaic virus, showing the great capability of this technique. However, the application of the TERS technique for a rapid and direct detection of different species of single viruses is under development, which is useful for a wide range of analytical fields.

Supported lipid structures and human cells (human dermal derived keratinocyte, HaCaT) were investigated using tip-enhancedRaman spectroscopy (TERS) to use the high spatial resolution capabilities of TERS, which is assumed to be less than 10 nm, to determine specific components on the cell surface. As lipids are a main component of cellular membranes, the correlation of spectral properties of pure lipids with respect to the complex biological sample was investigated. Induced by dynamic structural changes as well as nanoscale effects, a particular spectral feature of the lipid TERS spectra is found to vary, and a similar spectral deviation appears among the TERS spectra measured on the cell. Modifications of the cell surface alone cannot cause such behaviour. In contrast to soft lipid agglomerates, the cells were fixed and therefore hampered for intrinsic structural changes. Hence, the main contribution for the cell TERS spectra variation results from nanoscale effects, determined by different spectral characteristics compared to conventional Raman spectroscopy. The present results demonstrate the capability of TERS to provide a detailed and fast insight into the composition of the cell surface, even allowing the detection of single components.

In order to detect biomolecules, different approaches using for instance biological, spectroscopic or imaging techniques are established. Due to the broad variety of these methods, this review is focused on surface enhanced Raman spectroscopy (SERS) as an analytical tool in biomolecule detection. Here, the molecular specificity of Raman spectroscopy is combined with metallic nanoparticles as sensor platform, which enhances the signal intensity by several orders of magnitude. Within this article, the characterization of diverse biomolecules by means of SERS is explained and moreover current application fields are presented. The SERS intensity and as a consequence thereof the reliable detection of the biomolecule of interest is effected by distance, orientation and affinity of the molecule towards the metal surface. Furthermore, the great capability of the SERS technique for cutting-edge applications like pathogen detection and cancer diagnosis is highlighted. We wish to motivate by this comprehensive and critical summary researchers from various scientific background to create their own ideas and schemes for a SERS-based detection and analysis of biomolecules.

A first vibrational mapping on the nanometer scale was performed on a protein (streptavidin) labelled supported phospholipid film by means of tip-enhanced Raman spectroscopy (TERS). For this purpose a TERS spectral map was measured on the biomembrane model, using a step size far below the diffraction limit. Considering the model composition, spectra were classified as either typical for lipids, proteins or both simultaneously, in a qualitative manner. Subsequently, the spectroscopic information was assigned with respect to the topographic features. Since a spatial differentiation between different compositional domains is difficult to achieve by topographic features only, the combination of morphology and spectral data enables a much more detailed characterization of biomembranes.

New types of microfabricated surface-enhanced Raman spectroscopy (SERS) active substrates produced by electron beam lithography and ion beam etching are introduced. In order to achieve large enhancement factors by using the lightning rod effect, we prepare arrays consisting of sharp-edged nanostructures instead of the commonly used dots. Two experimental methods are used for fabrication: a one-stage process, leading to gold nanostar arrays and a two-stage process, leading to gold nanodiamond arrays. Our preparation process guarantees high reproducibility. The substrates contain a number of arrays for practical applications, each 200x200 microm2 in size. To test the SERS activity of these nanostar and nanodiamond arrays, a monolayer of the dye crystal violet is used. Enhancement factors are estimated to be at least 130 for the nanodiamond and 310 for the nanostar arrays.