Raman spectroscopy has been shown by various groups over the last two decades to have significant capability in discriminating disease states in bodily fluids, cells and tissues. Recent development in instrumentation, optics and manufacturing approaches has facilitated the design and demonstration of various novel in vivo probes, which have applicability for myriad of applications. This review focusses on key considerations and recommendations for application specific clinical Raman probe design and construction. Raman probes can be utilised as clinical tools able to provide rapid, non-invasive, real-time molecular analysis of disease specific changes in tissues. Clearly the target tissue location, the significiance of spectral changes with disease and the possible access routes to the region of interest will vary for each clinical application considered. This review provides insight into design and construction considerations, including suitable probe designs and manufacturing materials compatible with Raman spectroscopy.

The objective of this study is to use time-resolved (TR) Raman spectroscopy, spatially offset Raman spectroscopy (SORS), and a combination of these approaches to obtain high quality Raman spectra from materials hidden underneath an opaque layer. Both TR Raman and SORS are advanced techniques that allow for an increased relative selectivity of photons from deeper layers within a sample. Time-resolved detection reduces fluorescence background, and the selectivity for the second layer is improved. By combining this with spatially offset excitation we additionally increased selectivity for deeper layers. Test samples were opaque white polymer blocks of several mm thicknesses. Excitation was carried out with a frequency-doubled Ti:sapphire laser at 460 nm, 3 ps pulse width and 76 MHz repetition rate. Detection was either with a continuous-wave CCD camera or in time-resolved mode using an intensified CCD camera with a 250 ps gate width. The Raman photons were collected in backscatter mode, with or without lateral offset. By measuring the delay of the Raman signal from the second layer (polyethylene terephthalate/PET/Arnite), the net photon migration speeds through Teflon, polythene, Delrin and Nylon were determined. Raman spectra could be obtained from a second layer of PET through Teflon layers up to 7 mm of thickness. The ability to obtain chemical information through layers of diffusely scattering materials has powerful potential for biomedical applications.

The detection of explosives concealed behind opaque, diffusely scattering materials is a challenge that requires noninvasive analytical techniques for identification without having to manipulate the package. In this context, this study focuses on the application of time-resolved Raman spectroscopy (TRRS) with a picosecond pulsed laser and an intensified charge-coupled device (ICCD) detector for the noninvasive identification of explosive materials through several millimeters of opaque polymers or plastic packaging materials. By means of a short (250 ps) gate which can be delayed several hundred picoseconds after the laser pulse, the ICCD detector allows for the temporal discrimination between photons from the surface of a sample and those from deeper layers. TRRS was applied for the detection of the two main isomers of dinitrotoluene, 2,4-dinitrotoluene, and 2,6-dinitrotoluene as well as for various other components of explosive mixtures, including akardite II, diphenylamine, and ethyl centralite. Spectra were obtained through different diffuse scattering white polymer materials: polytetrafluoroethylene (PTFE), polyoxymethylene (POM), and polyethylene (PE). Common packaging materials of various thicknesses were also selected, including polystyrene (PS) and polyvinyl chloride (PVC). With the demonstration of the ability to detect concealed, explosives-related compounds through an opaque first layer, this study may have important applications in the security and forensic fields.

Time resolved Raman spectroscopy (TRRS) can provide subsurface information from multi-layered samples of transparent and translucent evaporative and silicate minerals up to several centimetres thick. Depth information was obtained using 3-ps pulsed laser excitation at 720 nm and a gated intensified charge-coupled device detector with stepwise increasing delay times. Blocks of different minerals were used as first, second or third layers, and Raman spectra from deeper layers could be detected through 10 mm of translucent calcite and up to 40 mm of transparent halite crystals. Measurements by conventional confocal Raman, as well as spatially offset Raman spectroscopy were also successful in distinguishing different mineral layers. This study establishes the great potential for the use of Raman spectroscopy in future planetary exploration, where TRRS could be used as a non-invasive tool for profiling the (sub-)surface at millimetre-depth resolution.

General rightsThis document is made available in accordance with publisher policies. Please cite only the published version using the reference above. AbstractHere we demonstrate the first use of a multifibre Raman probe that fits inside the bore of a hypodermic needle. A Raman probe utilising multiple collection fibres provides improved signal collection efficiency in biological samples compared with a previous 2-fibre design. Furthermore, probe performance (signal-to-noise ratios (SNRs)) compared favourably with those achieved in previous Raman microscope experiments able to distinguish benign, primary malignancies and secondary malignancies in lymph nodes.Experimental measurements presented in this paper give an indication of the sampling volume of the Raman needle probe in lymphoid tissues. Liquid tissue phantoms were used, containing scattering media encompassing a range of scattering properties similar to a variety of tissue types, including lymph node tissues. To validate the appropriateness of phantoms, the sampling depth of the probe was also measured in excised lymph node tissue. Greater than 50% of Raman photons collected were found to originate between the tip of the needle and a depth of 500 µm into the tissue.The needle probe presented here achieves comparable spectral quality to numerous studies previously demonstrating Raman disease discrimination. It is expected that this approach could achieve targeted subcutaneous tissue measurements and be viable for use for the in vivo Raman diagnostics of solid organs located within a few cm below the skin's surface.

Time-resolved and spatially offset Raman spectroscopies have previously been demonstrated for depth analysis through strongly scattering, non-transparent materials. In this study, several series of tissue phantoms were created with varied compositions and thicknesses to compare the potential of these different Raman techniques for biomedical applications. Polydimethylsiloxane (PDMS) phantoms were made with TiO2 particles suspended as a scattering agent, mimicking the scattering properties of biological tissues. The phantom layers contained embedded biomineral simulating inclusions (sphere or layer-shaped) with varied carbonate to phosphate ratios. The tissue phantoms were studied using Time Resolved Raman Spectroscopy (TRRS), Spatially Offset Raman Spectroscopy (SORS), and their combination, using a single instrumental setup with picosecond pulsed excitation at 720 nm and two different detectors. A comparison is made of the efficiency of these techniques to resolve chemical information from these heterogeneous scattering phantom samples. Measurements with continuous wave detection were found to offer a better signal-to-noise ratio than with TRRS, and in SORS measurements ratios of target to matrix signal were found to vary depending on the structural geometry and optical properties of the phantoms. Anomalous SORS behaviour, in which the relative contribution from the target decreases with offset, was observed in cases where the target was highly scattering and the top layer was relatively transparent. Time gating with an intensified charge-coupled device (ICCD) detector can yield more direct information on the depth of the hidden material.

Raman spectroscopy is one of the major characterization methods employed over the last few decades as a nondestructive technique for the study of heterogeneous catalysts and related catalytic reactions. However, the promise of practical applicability on millimeter-sized catalyst bodies, such as extrudates, has not been fulfilled completely. Large fluorescence signals and the highly scattering nature of the extrudates often hamper its practical usage. Different approaches to overcome this problem were examined, including the use of time-resolved Raman spectroscopy (TRRS), spatially offset Raman spectroscopy (SORS), surface-enhanced Raman spectroscopy (SERS), and combinations of these techniques. This paper demonstrates that especially TRRS can provide chemical information at depth within catalyst bodies, overcoming fluorescence background signals and allowing for visualization of analytes at different depths. It also examines the application of time-resolved SERS within catalyst bodies to gain insight into localized activity. With these options a wider applicability of Raman spectroscopy for industrial catalysis research becomes within reach.

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