Advanced Microspatially Offset Raman Spectroscopy for Noninvasive Imaging of Concealed Texts and Figures Using Raman Signal, Fluorescence Emission, and Overall Spectral Intensity.

This study explores the possibility of using microspatially offset Raman spectroscopy (micro-SORS) imaging to reconstruct noninvasively letters and figures hidden by opaque layers. Micro-SORS experiments were conducted on mockup samples that mimic real situations encountered in the cultural heritage field, such as sealed letters with inaccessible text and original documents. Subsurface images were obtained using both the characteristic Raman bands of the hidden compounds and their different optical properties from the remaining matrix. In the latter case, contrast obtained through observing a difference in the overall spectral intensity and fluorescence profile rather than any specific Raman bands were used to track the images within the hidden layer. This approach opens new prospects for the use of micro-SORS in heritage science, with applications in the field that include the study of objects covered by opaque overlayers not only through their Raman signatures but also through differences in their optical properties (e.g., fluorescence emission, absorption).

Non-destructive Monitoring of Dye Depth Profile in Mesoporous TiO2 Electrodes of Solar Cells with Micro-SORS.

The dye distribution within a photo-electrode is a key parameter in determining the performances of dye-sensitized photon-to-electron conversion devices, such as dye-sensitized solar cells (DSSCs). A traditional, depth profiling investigation by destructive means including cross-sectional sampling is unsuitable for large quality control applications in manufacturing processes. Therefore, a non-destructive monitoring of the dye depth profile is required, which is the first step toward a non-destructive evaluation of the internal degradation of the device in the field. Here, we present a conceptual demonstration of the ability to monitor the dye depth profile within the light active layer of DSSCs by non-destructive means with high chemical specificity using a recently developed non-destructive/non-invasive Raman method, micro-spatially offset Raman spectroscopy (micro-SORS). Micro-SORS is able to probe through turbid materials, providing the molecular identification of compounds located under the surface, without the need of resorting to a cross-sectional analysis. The study was performed on the photo-electrode of DSSCs. This represents the first demonstration of the micro-SORS concept in the solar cell area as well as, more generally, the application of micro-SORS to the thinnest layer to date. A sample set has been prepared with varying concentrations of the dye and the thickness of the matrix consisting of a titanium dioxide layer. The results showed that micro-SORS can unequivocally discriminate between the homogeneous and inhomogeneous dye depth profiles. Moreover, micro-SORS outcomes have been compared with the results obtained with destructive time-of-flight secondary ion mass spectrometry measurements. The results of the two techniques are in good agreement, confirming the reliability of micro-SORS analysis. Therefore, this study is expected to pave the way for establishing a wider and more effective monitoring capability in this important field.

Close to the diffraction limit in high resolution ATR FTIR mapping: demonstration on micrometric multi-layered art systems.

This paper is aimed at demonstrating the potentiality of high resolution Attenuated Total Reflection Fourier Transform Infrared micro-mapping (micro-ATR-FTIR) to reconstruct the images of micrometric multi-layered systems. This method can be an effective analytical alternative when the layer thickness requires high lateral resolution, and fluorescence or thermal effects prevent the deployment of conventional analytical techniques such as micro-Raman spectroscopy. This study demonstrates the high micro-ATR-FTIR setup performances in terms of lateral resolution, spectral quality and chemical image contrast using a new laboratory instrument equipped with a single element detector. The method has been first validated on mock-ups and then successfully applied on cross-sectional samples from real artworks: Leonardo da Vinci's mural painting, characterised by a few micrometers thin sequence of organic and inorganic layers, and an outdoor marble statue, with a complex sequence of decay products on its surface. This study paves the way to a new investigation modality of micrometric systems, combining high lateral resolution with excellent spectral quality, essential in the field of Cultural Heritage as well as in the wider area of materials and forensic sciences.

Investigation of Heterogeneous Painted Systems by Micro-Spatially Offset Raman Spectroscopy.

A recently developed technique of Micro-Spatially Offset Raman Spectroscopy (micro-SORS) extends the applicability of Raman spectroscopy to probing thin, highly diffusely scattering layers such as stratified paint samples, enabling their nondestructive chemical characterization. The technique has a wide applicability across areas such as cultural heritage, polymer research, forensics, and biological fields; however, currently, it suffers from a major unaddressed issue related to its ineffectiveness with highly heterogeneous samples. In this paper, we address this unmet need while demonstrating an effective strategy to probe such samples, involving a mapping on scales substantially larger than the scale of heterogeneity. This approach provides an effective means of obtaining robust and representative micro-SORS datasets from which sample composition can be effectively deduced, even in these extreme scenarios. The approach is compared with a basic point collection approach on two-layer paint systems where different layers-top, bottom, or both-are heterogeneous. The study has particular relevance to cultural heritage, where heterogeneous layers are often encountered with painted stratigraphies.

Discovering Hidden Painted Images: Subsurface Imaging Using Microscale Spatially Offset Raman Spectroscopy.

We demonstrate for the first time the mapping capability of micro-spatially offset Raman spectroscopy (micro-SORS). The technique enables to form noninvasive images of thin sublayers through highly turbid overlayers. The approach is conceptually demonstrated on recovering overpainted images in situations where conventional Raman microscopy was unable to visualize the sublayer. The specimens mimic real situations encountered in Cultural Heritage that deal, for example, with hidden paintings vandalized with graffiti or covered by superimposed painted layers or whitewash. Additionally, using a letter as a hidden image, we demonstrated the micro-SORS potential to reconstruct also a hidden writing covered, for example, with paper sheets that cannot be easily removed. Potential applications could also include other disciplines such as polymers, biological, catalytic, and forensic sciences where thin, highly turbid layers mask chemically distinct subsurface structures.

Development of a full micro-scale spatially offset Raman spectroscopy prototype as a portable analytical tool.

We present, for the first time, a portable full micro-Spatially Offset Raman Spectroscopy (micro-SORS) prototype permitting the in situ analysis of thin, highly turbid stratified layers at depths not accessible to conventional Raman microscopy. The technique is suitable for the characterisation of painted layers in panels, canvases and mural paintings, painted statues and decorated objects in cultural heritage or stratified polymers, and biological, catalytic and forensics samples where invasive analysis is undesirable or impossible to perform. The new device is characterised conceptually in polymer and paint layer systems. The provision of portability with full micro-SORS delivers subsurface micro-SORS capability unlocking the non-invasive and non-destructive potential of micro-SORS at its most effective form permitting it to be applied to large and non-portable objects in situ without recourse to removing micro-fragments for laboratory analysis on benchtop Raman microscopes.

Determination of thickness of thin turbid painted over-layers using micro-scale spatially offset Raman spectroscopy.

We present a method for estimating the thickness of thin turbid layers using defocusing micro-spatially offset Raman spectroscopy (micro-SORS). The approach, applicable to highly turbid systems, enables one to predict depths in excess of those accessible with conventional Raman microscopy. The technique can be used, for example, to establish the paint layer thickness on cultural heritage objects, such as panel canvases, mural paintings, painted statues and decorated objects. Other applications include analysis in polymer, biological and biomedical disciplines, catalytic and forensics sciences where highly turbid overlayers are often present and where invasive probing may not be possible or is undesirable. The method comprises two stages: (i) a calibration step for training the method on a well characterized sample set with a known thickness, and (ii) a prediction step where the prediction of layer thickness is carried out non-invasively on samples of unknown thickness of the same chemical and physical make up as the calibration set. An illustrative example of a practical deployment of this method is the analysis of larger areas of paintings. In this case, first, a calibration would be performed on a fragment of painting of a known thickness (e.g. derived from cross-sectional analysis) and subsequently the analysis of thickness across larger areas of painting could then be carried out non-invasively. The performance of the method is compared with that of the more established optical coherence tomography (OCT) technique on identical sample set.This article is part of the themed issue 'Raman spectroscopy in art and archaeology'.

Fluorescence suppression using micro-scale spatially offset Raman spectroscopy.

We present a new concept of fluorescence suppression in Raman microscopy based on micro-spatially offset Raman spectroscopy which is applicable to thin stratified turbid (diffusely scattering) matrices permitting the retrieval of the Raman signals of sublayers below intensely fluorescing turbid over-layers. The method is demonstrated to yield good quality Raman spectra with dramatically suppressed fluorescence backgrounds enabling the retrieval of Raman sublayer signals even in situations where conventional Raman microscopy spectra are fully overwhelmed by intense fluorescence. The concept performance was studied theoretically using Monte Carlo simulations indicating the potential of up to an order or two of magnitude suppression of overlayer fluorescence backgrounds relative to the Raman sublayer signals. The technique applicability was conceptually demonstrated on layered samples involving paints, polymers and stones yielding fluorescence suppression factors between 12 to above 430. The technique has potential applications in a number of analytical areas including cultural heritage, archaeology, polymers, food, pharmaceutical, biological, biomedical, forensics and catalytic sciences and quality control in manufacture.

Portable Sequentially Shifted Excitation Raman spectroscopy as an innovative tool for in situ chemical interrogation of painted surfaces.

We present the first validation and application of portable Sequentially Shifted Excitation (SSE) Raman spectroscopy for the survey of painted layers in art. The method enables the acquisition of shifted Raman spectra and the recovery of the spectral data through the application of a suitable reconstruction algorithm. The technique has a great potentiality in art where commonly a strong fluorescence obscures the Raman signal of the target, especially when conventional portable Raman spectrometers are used for in situ analyses. Firstly, the analytical capability of portable SSE Raman spectroscopy is critically discussed using reference materials and laboratory specimens, comparing its results with other conventional high performance laboratory instruments (benchtop FT-Raman and dispersive Raman spectrometers with an external fiber optic probe); secondly, it is applied directly in situ to study the complex polychromy of Italian prestigious terracotta sculptures of the 16(th) century. Portable SSE Raman spectroscopy represents a new investigation modality in art, expanding the portfolio of non-invasive, chemically specific analytical tools.

Development of portable defocusing micro-scale spatially offset Raman spectroscopy.

We present, for the first time, portable defocusing micro-Spatially Offset Raman Spectroscopy (micro-SORS). Micro-SORS is a concept permitting the analysis of thin, highly turbid stratified layers beyond the reach of conventional Raman microscopy. The technique is applicable to the analysis of painted layers in cultural heritage (panels, canvases and mural paintings, painted statues and decorated objects in general) as well as in many other areas including polymer, biological and biomedical applications, catalytic and forensics sciences where highly turbid stratified layers are present and where invasive analysis is undesirable or impossible. So far the technique has been demonstrated only on benchtop Raman microscopes precluding the non-invasive analysis of larger samples and samples in situ. The new set-up is characterised conceptually on a range of artificially assembled two-layer systems demonstrating its benefits and performance across several application areas. These included stratified polymer sample, pharmaceutical tablet and layered paint samples. The same samples were also analysed by a high performance (non-portable) benchtop Raman microscope to provide benchmarking against our earlier research. The realisation of the vision of delivering portability to micro-SORS has a transformative potential spanning across multiple disciplines as it fully unlocks, for the first time, the non-invasive and non-destructive aspects of micro-SORS enabling it to be applied also to large and non-portable samples in situ without recourse to removing samples, or their fragments, for laboratory analysis on benchtop Raman microscopes.

Analytical Capability of Defocused µ-SORS in the Chemical Interrogation of Thin Turbid Painted Layers.

A recently developed micrometer-scale spatially offset Raman spectroscopy (µ-SORS) method provides a new analytical capability for investigating non-destructively the chemical composition of sub-surface, micrometer-scale thickness, diffusely scattering layers at depths beyond the reach of conventional confocal Raman microscopy. Here, we demonstrate experimentally, for the first time, the capability of µ-SORS to determine whether two detected chemical components originate from two separate layers or whether the two components are mixed together in a single layer. Such information is important in a number of areas, including conservation of cultural heritage objects, and is not available, for highly turbid media, from conventional Raman microscopy, where axial (confocal) scanning is not possible due to an inability to facilitate direct imaging within the highly scattering sample. This application constitutes an additional capability for µ-SORS in addition to its basic capacity to determine the overall chemical make-up of layers in a turbid system.

Noninvasive analysis of thin turbid layers using microscale spatially offset Raman spectroscopy.

Here, we demonstrate, for the first time, the extension of applicability of recently developed microscale spatially offset Raman spectroscopy (SORS), micro-SORS, from the area of cultural heritage to a wider range of analytical problems involving thin, tens of micrometers thick diffusely scattering turbid layers. The method can be applied in situations where a high turbidity of layers prevents the deployment of conventional confocal Raman microscopy with its depth resolving capability. The method was applied successfully to detect noninvasively the presence of thin, highly turbid layers within polymers, wheat seeds, and paper. An invasive, cross sectional analysis confirmed the micro-SORS findings. Micro-SORS represents a new Raman imaging modality expanding the portfolio of noninvasive, chemically specific analytical tools.