In this paper, it is demonstrated how Raman spectroscopy can be used to detect different carotenoids as possible biomarkers in various groups of microorganisms. The question which arose from previous studies concerns the level of unambiguity of discriminating carotenoids using common Raman microspectrometers. A series of laboratory-grown microorganisms of different taxonomic affiliation was investigated, such as halophilic heterotrophic bacteria, cyanobacteria, the anoxygenic phototrophs, the non-halophilic heterotrophs as well as eukaryotes (Ochrophyta, Rhodophyta and Chlorophyta). The data presented show that Raman spectroscopy is a suitable tool to assess the presence of carotenoids of these organisms in cultures. Comparison is made with the high-performance liquid chromatography approach of analysing pigments in extracts. Direct measurements on cultures provide fast and reliable identification of the pigments. Some of the carotenoids studied are proposed as tracers for halophiles, in contrast with others which can be considered as biomarkers of other genera. The limits of application of Raman spectroscopy are discussed for a few cases where the current Raman spectroscopic approach does not allow discriminating structurally very similar carotenoids. The database reported can be used for applications in geobiology and exobiology for the detection of pigment signals in natural settings.

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Since evaporitic rocks on the Martian surface could (or still can) serve as potential habitats for microbial life on Mars, there is a reasonable possibility that these rocks may sustain molecular remnants as evidence for the presence of extinct or extant living organisms on Mars and that beta-carotene could be a suitable biomarker. In this paper, Raman microspectrometry was tested as a nondestructive method of determining the lowest detectable beta-carotene content in experimentally prepared evaporitic matrices--namely, gypsum, halite and epsomite. Two excitation wavelengths were compared--514.5 nm, because of the resonance Raman enhancement in the carotenoid analysis, and 785 nm, as a more universal wavelength now much used in the detection of biomolecules terrestrially. Mixtures were measured directly as well as with a laser beam penetrating the crystals of gypsum and epsomite. We have obtained beta-carotene signals at the 0.1 to 10 mg kg(-1) level--the number of registered beta-carotene Raman bands differed depending on the particular mineral matrix and the excitation wavelength. Concentrations of beta-carotene of about one order of magnitude higher were identified when analysed through single crystals of gypsum and epsomite, respectively.

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Flexirubins are specific polyene pigments produced by several genera of Bacteroidetes. Colonies and cell extracts of Flavobacterium johnsoniae and Flexibacter elegans have been investigated by Raman spectroscopy to show that this fast and non-destructive technique can be used to differentiate these pigments from carotenoids and to compare the flexirubin content of the two microorganisms. The presence or absence of certain distinguishing features in the CH combination band region at 2500-2750 cm(-1) can assist in the discrimination between the two flexirubins investigated. Raman spectroscopy is thus a suitable tool not only to detect flexirubin pigments in bacterial cells, but also to further characterize the pigments present in members of the Bacteroidetes genera that are rich in flexirubins.

A prototype instrument, under development at the University of Leicester, for the future European Space Agency (ESA) ExoMars mission, was used for the analysis of microbial pigments within a stratified gypsum crust from a hypersaline saltern evaporation pond at Eilat (Israel). Additionally, the same samples were analysed using a miniaturized Raman spectrometer, featuring the same 532 nm excitation. The differences in the position of the specific bands, attributed to carotenoid pigments from different coloured layers, were minor when analysed by the ESA prototype instrument; therefore, making it difficult to distinguish among the different pigments. The portable Delta Nu Advantage instrument allowed for the discrimination of microbial carotenoids from the orange/green and purple layers. The purpose of this study was to complement previous laboratory results with new data and experience with portable or handheld Raman systems, even with a dedicated prototype Raman system for the exploration of Mars. The latter is equipped with an excitation wavelength falling within the carotenoid polyene resonance region. The ESA prototype Raman instrument detected the carotenoid pigments (biomarkers) with ease, although further detailed distinctions among them were not achieved.

Vivianite (Fe3(PO4)2·8H2O) is a rare blue historical pigment, which can be profitably used in authorship ascription or copy identification. However, its tendency to degrade complicates its proper identification in paint layers. Reference vivianite mineralogical samples were analysed in order to compare and to test the limits of structural analyses on possibly degraded vivianite samples (X‐ray diffraction and vibrational spectroscopies). The same methods, in their μ‐configuration, were tested on micro‐samples of the paintings and their limits evaluated. A sedimentary origin of the pigment has been suggested. Vivianite was detected in various works by Jean George de Hamilton (1672–1737) and in a Late Gothic Transylvanian altarpiece.

A silver Torah shield fitted with a set of precious stones and glass imitations crafted in Poland in the first half of the 19th century was investigated using two of the currently distributed portable and relatively low-cost Raman spectrometers in situ at the Jewish Museum in Prague. Observed Raman peaks corresponded well (+/À 3 cm À1 ) to the reference values. The hand-held instruments operated at 785-and 532-nm laser excitations showed good performance in the fast and unambiguous identification of nearly 60 stones which were fitted on the shield: one blue aquamarine, three purple amethysts, thirteen red garnets (all classified as high-percentage almandines), three white pearls, fifteen pieces of red coral and five chalcedonies (one white and four red). All of the other stones were identified as colored glass. The rather chaotic mixture of stones of various colors, cuts and sizes and the total volume of imitation glass support the theory that the mounted stones were gathered from Jewish households and donated for the adornment of the shield. The common portable Raman instruments represent an ideal tool for the quick and accurate identification of gemstones mounted in historical artifacts in situ in the framework of museum or collection sites in a non-destructive way.