Imaging and micro-invasive analyses of black stains on the passepartout of Codex Atlanticus Folio 843 by Leonardo da Vinci.

This paper accounts for the diagnostic campaign aimed at understanding the phenomenon of black stains appeared on the passepartout close to the margins of Folio 843 of Leonardo da Vinci's Codex Atlanticus. Previous studies excluded microbiological deterioration processes. The study is based on a multi-analytical approach, including non-invasive imaging measurements of the folio, micro-imaging and synchrotron spectroscopy investigations of passepartout fragments at different magnifications and spectral ranges. Photoluminescence hyperspectral and lifetime imaging highlighted that black stains are not composed of fluorescent materials. µATR-FTIR imaging of fragments from the passepartout revealed the presence of a mixture of starch and PVAc glues localized only in the stained areas close to the margin of the folio. FE-SEM observations showed that the dark stains are localized inside cavities formed among cellulose fibers, where an accumulation of inorganic roundish particles (∅100-200 nm in diameter size), composed of Hg and S, was detected. Finally, by employing synchrotron µXRF, µXANES and HR-XRD analyses it was possible to identify these particles as metacinnabar (ß-HgS). Further research is needed to assess the chemical process leading to the metacinnabar formation in the controlled conservation condition of Leonardo's Codex.

Photoluminescence excited at variable fluences: a novel approach for studying the emission from crystalline pigments in paints.

Crystalline solids can exhibit photoluminescence when properly excited by sufficiently energetic light radiation. Following excitation, different radiative and non-radiative recombination pathways can occur that are informative of the energetic structure of the material as well as of the presence of crystal defects and impurities. Usually, the characterization of the optical emission of crystalline materials is achieved through the study of emission spectra as a function of the excitation wavelength. A different approach employs variable excitation fluence to populate the energetic levels until saturation, which promotes the emission from other radiative and non-radiative pathways. The method is particularly effective for understanding conduction phenomena and studying charge recombination channels in semiconductor materials. In this work, we propose its application for characterizing radiative recombination paths in crystalline pigments. The approach has been tested in spectroscopy mode for the identification of paints in a model painting and in micro-imaging modality for the study of paint stratigraphies. We demonstrate that the method is highly informative of the nature of different recombination paths in crystalline pigments and allows a deeper characterization of the emission from luminescent paints with respect to the conventional steady-state photoluminescence approach.

Effects of cleaning procedures on the long-term corrosion behavior of bronze artifacts of the cultural heritage in outdoor environment.

The cleaning of metallic artworks is a crucial step for their preservation. Cleaning operations generally aim at removing deposits and corrosion layers, and all the non-stable and potentially reactive phases formed as a consequence of the interaction of the metal with the environment. Thus, all secondary-formed compounds and layers that can undermine the overall preservation of the artwork, both from the esthetic and the corrosion point of view, should be removed. On the other hand, superficial stable patinas contributing to the artistic and historic value of the objects and that may provide protection to the metallic surface should be preserved. The optimal cleaning procedure should be able to promote a long-term improvement of the corrosion resistance of the surfaces. Therefore, the long-term monitoring of the corrosion behavior of the cleaned surfaces with electrochemical techniques could provide valuable information for the selection of the optimal methodology. In this work, five cleaning procedures have been applied to five bronze sculptures. The cleaned surfaces have been characterized following a multi-analytical and non-invasive approach, and the long-term evolution of their corrosion behavior has been monitored by means of on-site non-invasive linear polarization resistance (LPR) and electrochemical impedance spectroscopy (EIS) measurements for more than 2 years.

Stone/Coating Interaction and Durability of Si-Based Photocatalytic Nanocomposites Applied to Porous Lithotypes.

The use of hybrid nanocoatings for the protection of natural stones has received increasing attention over the last years. However, the interaction of these materials with stones and, in particular, its modification resulting from the blending of nanoparticles and matrices, are yet little explored. In this work, the interaction of two nanocomposite coatings (based on alkylalkoxysilane matrices and TiO2 nanoparticles in water and 2-propanol) with two different porous stones is examined in detail by comparing their absorption behaviour and protection performance with those of the respective TiO2-free matrices. It is shown that the protective effectiveness of both matrices is not negatively affected by the presence of TiO2, as the desired water barrier effect is retained, while a significant photocatalytic activity is achieved. The addition of titania leads to a partial aggregation of the water-based matrix and accordingly reduces the product penetration into stones. On the positive side, a chemical interaction between titania and this matrix is observed, probably resulting in a greater stability of nanoparticles inside the protective coating. Moreover, although an effect of TiO2 on the chemical stability of matrices is observed upon UV light exposure, the protective performance of coatings is substantially maintained after ageing, while the interaction between matrices and nanoparticles results in a good retention of the latter upon in-lab simulated rain wash-out.

Time-Resolved Photoluminescence Microscopy Combined with X-ray Analyses and Raman Spectroscopy Sheds Light on the Imperfect Synthesis of Historical Cadmium Pigments.

Recent studies have shown that modern pigments produced after the Second Industrial Revolution are complex systems characterized by a high level of heterogeneities. Therefore, it is fundamental to adopt a multianalytical approach and highly sensitive methods to characterize the impurities present within pigments. In this work we propose time-resolved and spectrally resolved photoluminescence (PL) microscopy for the mapping of luminescent crystal defects and impurities in historical cadmium-based pigments. PL analysis is complemented by X-ray diffraction, X-ray fluorescence and Raman spectroscopies, and by scanning electron microscopy to determine the chemical composition and crystal structure of samples. The study highlights the heterogeneous and complex nature of historical samples that can be associated with the imperfect manufacturing processes tested during the period between the 1850s and 1950s. The results also allow us to speculate on a range of synthesis processes. Since it is recognized that the stability of paints can be related to pigments synthesis, this research paves the way to a wider study on the relationship between synthesis methods and deterioration of cadmium pigments and paints. This rapid and immediate approach using PL can be applied to other semiconductor pigments and real case studies.

Biofilm colonization of metamorphic lithotypes of a renaissance cathedral exposed to urban atmosphere.

Stone architectural heritage exposed outdoor represents a challenging habitat for biological growths; nevertheless, biocolonization on heritage structure is ubiquitous and represents a major mechanism of alteration. However, the identification of specific microorganisms with known reactivity towards the stone substrate does not necessarily imply that a biodeterioration process is in progress and, in specific conditions, bioprotection effects have been highlighted as a result of colonization. The main objective of the present research is to evaluate the biofilm formation on different lithotypes exposed to similar environmental polluted conditions, and to investigate whether the presence of subaerial biofilms can be associated to an increased magnitude of deterioration of the colonized surfaces with respect to the not colonized ones. In particular, the research examines the extensive biological colonization of the stone surfaces of the façade of the Cathedral of Monza (Italy). Four metamorphic stones widely used in the façade and showing rather different compositional, mineralogical and microstructural features were studied. The state of conservation of the stones was characterized under the mineralogical and compositional point of view by X-ray diffraction and Fourier Transformed infrared analysis. The microstructure of colonized substrates and of reference not colonized ones was studied by means of optical and electron microscopy, to comparatively evaluate the damage extent and weathering patterns in both conservative conditions. The structure and the architecture of biofilms growing on different lithic surfaces were investigated by CLSM in both fluorescence and reflection modes. Captured images were analyzed for 3D reconstructions of biofilm samples. The biovolumes were also calculated to estimate the total biomass. The results indicate that the four lithotypes showed different colonization extents. However, even in presence of extensive biological growth, chemical-physical deterioration mechanisms caused by environmental exposure were largely responsible for deterioration. A relationship between compositional and surface morphological features and biocolonization was also observed.

Layered Nano-TiO2 Based Treatments for the Maintenance of Natural Stones in Historical Architecture.

Layered treatments of natural stones based on dispersions of experimental nano-TiO2 and commercial TEOS showing photocatalytic and self-cleaning properties were set up and tested. To enhance nano-TiO2 efficacy, a surface pre-treatment with tetraethyl orthosilicate was proposed to avoid the penetration of NPs into the crystalline porous substrates and to improve their adhesion to the stone. Two treatment applications (wet-on-wet and wet-on-dry) were compared, showing different results. A strong interaction Si-O-Ti was the key factor for the successful treatment, leaving the band gap and relevant properties of nano-TiO2 unaltered. The layered treatments were tested on a porous calcarenite (Noto stone) and a very compact marble (Carrara marble). The combined SiO2 -nano-TiO2 treatments can find application in suitable cases where a surface consolidation is needed, ensuring a depolluting and self-cleaning durable activity.

Photocatalytic Nanocomposites for the Protection of European Architectural Heritage.

In the field of stone protection, the introduction of inorganic nanoparticles, such as TiO2, ZnO, and Ag in polymeric blends can enhance the protective action of pristine treatments, as well as confer additional properties (photocatalytic, antifouling, and antibacterial). In the framework of the "Nano-Cathedral" European project, nanostructured photocatalytic protective treatments were formulated by using different TiO2 nanoparticles, solvents, and silane/siloxane systems in the blends. The results about the characterization and application of two promising nano-TiO2 based products applied on Apuan marble and Ajarte limestone are here reported, aiming at investigating the complex system "treatment/stone-substrate". The nanocomposites show better performances when compared to a commercial reference siloxane based protective treatment, resulting in different performances once applied on different carbonatic substrates, with very low and high open porosity, confirming the necessity of correlating precisely the characteristics of the stone material to those of the protective formulations. In particular, the TiO2 photocatalytic behavior is strictly linked to the amount of available nanoparticles and to the active surface area. The alkyl silane oligomers of the water-based formulation have a good penetration into the microstructure of Ajarte limestone, whereas the solvent-based and small size monomeric formulation shows better results for Apuan marble, granting a good coverage of the pores. The encouraging results obtained so far in lab will be confirmed by monitoring tests aiming at assessing the effectiveness of the treatments applied in pilot sites of historical Gothic Cathedrals.

Synchrotron-based FTIR microspectroscopy for the mapping of photo-oxidation and additives in acrylonitrile-butadiene-styrene model samples and historical objects.

Synchrotron-based Fourier transform infrared micro-spectroscopy (SR-µFTIR) was used to map photo-oxidative degradation of acrylonitrile-butadiene-styrene (ABS) and to investigate the presence and the migration of additives in historical samples from important Italian design objects. High resolution (3×3 µm(2)) molecular maps were obtained by FTIR microspectroscopy in transmission mode, using a new method for the preparation of polymer thin sections. The depth of photo-oxidation in samples was evaluated and accompanied by the formation of ketones, aldehydes, esters, and unsaturated carbonyl compounds. This study demonstrates selective surface oxidation and a probable passivation of material against further degradation. In polymer fragments from design objects made of ABS from the 1960s, UV-stabilizers were detected and mapped, and microscopic inclusions of proteinaceous material were identified and mapped for the first time.

Setup of galvanic sensors for the monitoring of gilded bronzes.

Traditional electrochemical techniques, such as linear polarization resistance (Rp), and electrochemical impedance spectroscopy (EIS), cannot be applied to gilded bronzes, as it may not be possible to interpret the results obtained due to the bimetallic nature of the studied material. The measurement of the macrocouple current generated by the gold/bronze galvanic couple can be used as an indicator of degradation processes. Nevertheless, this measurement cannot be performed directly on the original artifacts due to the systematic presence of short-circuits between the two metals. In the present work the use of galvanic sensors is proposed as a possible solution for the monitoring of gilded bronze artefacts. The sensors have been designed to simulate real gilded bronze surfaces in terms of composition and stratigraphy and have proved to be a reliable diagnostic tool for the in situ monitoring of the rates of deterioration of gilded bronze surfaces and to test new conservation treatments. Their set-up and application is reported and their performances discussed.

Time-resolved photoluminescence spectroscopy and imaging: new approaches to the analysis of cultural heritage and its degradation.

Applications of time-resolved photoluminescence spectroscopy (TRPL) and fluorescence lifetime imaging (FLIM) to the analysis of cultural heritage are presented. Examples range from historic wall paintings and stone sculptures to 20th century iconic design objects. A detailed description of the instrumentation developed and employed for analysis in the laboratory or in situ is given. Both instruments rely on a pulsed laser source coupled to a gated detection system, but differ in the type of information they provide. Applications of FLIM to the analysis of model samples and for the in-situ monitoring of works of art range from the analysis of organic materials and pigments in wall paintings, the detection of trace organic substances on stone sculptures, to the mapping of luminescence in late 19th century paintings. TRPL and FLIM are employed as sensors for the detection of the degradation of design objects made in plastic. Applications and avenues for future research are suggested.

A novel classification method for multispectral imaging combined with portable Raman spectroscopy for the analysis of a painting by Vincent Van Gogh.

In this work, a novel combination of portable micro-Raman spectroscopy and semi-automatic methods of data treatment are proposed for the classification and mapping of visible multispectral imaging data for the analysis of a painting on paper by Vincent Van Gogh. Analysis of multispectral imaging data with the sequential maximum-angle convex cone (SMACC) and spectral angle mapper (SAM) algorithms differentiated the surface into areas on the basis of the presence of pigment mixtures. Complementary analytical information was obtained through portable Raman spectroscopy was performed on a few selected points of the painting, allowing for the determination of Van Gogh's palette and the mapping of pigment mixtures on the painting's surface; the number of mixtures employed is varied and at least two different blues are present. The results obtained were integrated with the information from prior ultraviolet (UV)-induced luminescence analysis performed on the same painting to better understand the materials used by the artist. The mathematical treatment of multispectral data using the proposed methods could be extended to the analysis of other painted surfaces.

Importance of subaerial biofilms and airborne microflora in the deterioration of stonework: a molecular study.

The study characterized the sessile microbial communities on mortar and stone in Milan University's Richini's Courtyard and investigated the relationship between airborne and surface-associated microbial communities. Active colonization was found in three locations: green and black patinas were present on mortar and black spots on stone. Confocal laser scanning microscopy, scanning electron microscopy and culture-independent molecular methods revealed that the biofilm causing deterioration was dominated by green algae and black fungi. The mortar used for restoration contained acrylic and siloxane resins that could be used by microorganisms as carbon and energy sources thereby causing proliferation of the biofilm. Epifluorescence microscopy and culture-based methods highlighted a variety of airborne microflora. Bacterial and fungal counts were quantitatively similar to those reported in other investigations of urban areas, the exception being fungi during summer (1-2 orders of magnitude higher). For the first time in the cultural heritage field, culture-independent molecular methods were used to resolve the structure of airborne communities near discoloured surfaces, and to investigate the relationship between such communities and surface-associated biofilms.

The Silk Road, Marco Polo, a Bible and its proteome: a detective story.

Around the end of XIII century (at the time of young Marco Polo's first trip to China at the court of Khubilai Khan in Khan Baliq) a pocket Bible was delivered by a Franciscan friar to the Mogul Emperor, in the framework of the evangelization program of the Far East. Four centuries later, in 1685, this Bible was rediscovered by the Jesuit Philippe Couplet in the house of a rich Chinese in Nanchin and donated to Cosimo III, Grand Duke of Tuscany. This Bible was recently "unearthed" in the Biblioteca Medicea Laurenziana in Florence, wrapped up in a precious yellow silk cloth, in a rather ruined state. After two years of restoration, the Bible will return to China in 2012 for a celebration of its >700years of life and of its remarkable return trip on the Silk Road. On account of the thinness of the parchment (barely 80µm thickness, the size of each foil being 16.5×11cm) it was widely held that the pages were produced from foetal lambskins. On tiny fragments of the margins of a foil, after several unsuccessful attempts at digesting the vellum, we were able to obtain a tryptic peptide mixture, which, upon mass spectrometry analysis, yielded the identity of 8 unique proteins, belonging to the genus Bos taurus, thus confirming the origin of the vellum from calfskins rather than from foetal lambskins. Our results prove that it is possible to obtain reliable protein extraction and IDs from ancient parchment documents.

Fluorescence and Fourier-transform infrared spectroscopy for the analysis of iconic Italian design lamps made of polymeric materials.

The preservation of design object collections requires an understanding of their constituent materials which are often polymeric blends. Challenges associated with aging of complex polymers from objects with an unknown physical history may compromise the interpretation of data from analytical techniques, and therefore complicate the assessment of the condition of polymers in indoor museum environments. This study focuses on the analysis of polymeric materials from three well-known Italian design lamps from the 1960s. To assess the degree of chemical modifications in the polymers, non-destructive molecular spectroscopic techniques, Fourier-transform infrared (FTIR) and fluorescence spectroscopy, have been applied directly on the object surfaces using an optical fiber probe and through examination of micro samples. FTIR spectra of the different polymers, polyvinylacetate (PVAc) for the lamps Taraxacum and Fantasma, and both acrylonitrile-butadiene-styrene polymer (ABS) and cellulose acetate (CA) for the lamp Nesso, allowed the detection of ongoing deterioration processes. Fluorescence spectroscopy proved particularly sensitive for the detection of molecular changes in the polymeric objects, as the spectra obtained from the examined lamps differ significantly from those of the unaged reference materials. Differences in fluorescence spectra are also detected between different points on the same object further indicating the presence of different chemical species on the surfaces. With the aid of complementary data from FTIR spectroscopy, an interpretation of the emission spectra of the studied polymeric objects is here proposed, further suggesting that fluorescence spectroscopy may be useful for following the degradation of historical polymeric objects.

Assessment of the ageing of triterpenoid paint varnishes using fluorescence, Raman and FTIR spectroscopy.

The assessment of the influence of natural and artificial ageing on the spectrofluorescence of triterpenoid varnishes dammar and mastic is the focus of this work. Both Fourier transform infrared (FTIR) microscopy using attenuated total reflectance and Raman spectroscopy have been employed for complementary molecular analysis of samples. Synchronous fluorescence spectroscopy, excitation emission spectroscopy, and statistical analysis of data have been used to monitor changes in the optical properties of varnish samples. Assessment of naturally and artificially aged samples using excitation emission spectroscopy suggests that extensive exposure to visible light does not lead to easily appreciable differences in the fluorescence of mastic and dammar; cluster analysis has been used to assess changes, which occur with artificial ageing under visible light, indicating that differences in the fluorescence spectra of aged triterpenoids may be insufficient for their discrimination. The results highlight significant differences between the initial fluorescence of films of dammar and mastic and the fluorescence, which develops with ageing and oxidation, and specific markers, which change with ageing in FTIR and Raman spectra, have been identified.

N-vanillylnonanamide tested as a non-toxic antifoulant, applied to surfaces in a polyurethane coating.

The potential on N-vanillylnonanamide (NVN) in preventing the attachment of Pseudomonas stutzeri and a Bacillus cereus-group strain was investigated. NVN up to 852 microM was not toxic, nor was it an energy source for either organism. Microbial attachment assays were carried out on glass and polylysine slides. with NVN being dispersed in or applied to the surfaces using a polyurethane coating. NVN at 205 microM inhibited Bacillus adhesion on glass slides by 48% and the percentage did not significantly increase at 852 microM. NVN blended into or sprayed onto the coating at 205 micromol/kg did not prevent adhesion. The compound is therefore not useful as an antifouling product under the tested coating conditions.

Black layers on historical architecture.

BACKGROUND, AIM AND SCOPE: The external surface of any building in urban polluted environment is unavoidably destined to be covered with layers that assume a grey to black colour and are generally called 'black crusts'. These, according to standard protocols and glossary, are deteriorated surface layers of stone material; they can have variable thickness, are hard and fragile and can detach spontaneously from the substrate, which, in general, is quite decayed. Plain visual examination may lead to consider 'black crusts' all similar, whilst only a careful diagnostic investigation can distinguish 'black crusts' and the consequences of their formation on stone substrates. In this paper, various black layers on marble are studied and compared and the morphological and compositional characteristics discussed according to the related mechanisms of formation. Differences between old (hundred years) and recent crusts (30 years) are investigated and pointed out. MATERIALS AND METHODS: Samples of black crusts collected from the Milan Cathedral façade (Candoglia Marble) have been studied and compared with the careful and synergic employ of traditional techniques: optical (transmission and reflected VIS light) and electron microscopy, X-ray spectrometry and micro-Fourier transform infrared spectroscopy. RESULTS: Visual examination of loose fragments does not allow to point out outstanding differences amongst the various samples; black layers have similar main mineral components, gypsum and airborne particles, with different spatial distribution. The microscopic studies allowed to point out the porosity differences, the gypsum crystallisation habit, different amount of embedded particles, level and progress of marble decay. DISCUSSION: The observations lead to define three main types of black crusts: black crust deriving from marble sulphation, compact deposit and encrustation due to exogenic materials deposition. Black crusts show evidence of sulphation in progress, without a clear continuity solution between crust and marble; the lack of separation is particularly evident in 'recent' crust, where the sulphation process is more active. Black compact deposits show a higher porosity than black crusts because gypsum is not coming from the chemical corrosion of the substrate but from outside; actually, in the former case, the substrate is sound. Encrustations show a highly regular crystal organisation of gypsum (close packed tabular crystals) that cannot be traced back to casual atmospheric deposit or to corrosion of the substrate but rather to the crystallisation of a solution coming from an external source. Also in this case, the marble is sound; evidence of the effect of some protection treatment is pointed out. CONCLUSIONS: In spite of the apparent similarity of the examined samples, analytical results have evidenced three main types of black crusts: black crust with decayed substrate, compact deposit and black encrustation showing a sound substrate underneath. Experimental evidence of calcite grains sulphation in progress, taking place according to a model recently proposed, has been observed. Sulphation process is prevented where particular conservation treatments had been applied in the past. RECOMMENDATIONS AND PERSPECTIVES: New experimental studies can be focussed to understand the specific conditions (measurements of micro-climatic and thermodynamic parameters) and mechanisms for black crusts formation in situ. The problem of the kinetic of the sulphation process of marble, the assessment of black layers formation in the case of different carbonate stone materials and the study of acid attack in presence of surface protecting layers deserve further investigation.

A portable UV-fluorescence multispectral imaging system for the analysis of painted surfaces.

A portable fluorescence multispectral imaging system was developed and has been used for the analysis of artistic surfaces. The imaging apparatus exploits two UV lamps for fluorescence excitation and a liquid crystal tunable filter coupled to a low-noise charge coupled device as the image detector. The main features of the system are critically presented, outlining the assets, drawbacks, and practical considerations of portability. A multivariate statistical treatment of spectral data is further considered. Finally, the in situ analysis with the new apparatus of recently restored Renaissance wall paintings is presented.

Advantages of using microbial technology over traditional chemical technology in removal of black crusts from stone surfaces of historical monuments.

This study compares two cleaning methods, one involving an ammonium carbonate-EDTA mixture and the other involving the sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris ATCC 29579, for the removal of black crust (containing gypsum) on marble of the Milan Cathedral (Italy). In contrast to the chemical cleaning method, the biological procedure resulted in more homogeneous removal of the surface deposits and preserved the patina noble under the black crust. Whereas both of the treatments converted gypsum to calcite, allowing consolidation, the chemical treatment also formed undesirable sodium sulfate.