Identification of morphological biosignatures in Martian analogue field specimens using in situ planetary instrumentation.

We have investigated how morphological biosignatures (i.e., features related to life) might be identified with an array of viable instruments within the framework of robotic planetary surface operations at Mars. This is the first time such an integrated lab-based study has been conducted that incorporates space-qualified instrumentation designed for combined in situ imaging, analysis, and geotechnics (sampling). Specimens were selected on the basis of feature morphology, scale, and analogy to Mars rocks. Two types of morphological criteria were considered: potential signatures of extinct life (fossilized microbial filaments) and of extant life (crypto-chasmoendolithic microorganisms). The materials originated from a variety of topical martian analogue localities on Earth, including impact craters, high-latitude deserts, and hydrothermal deposits. Our in situ payload included a stereo camera, microscope, Mössbauer spectrometer, and sampling device (all space-qualified units from Beagle 2), and an array of commercial instruments, including a multi-spectral imager, an X-ray spectrometer (calibrated to the Beagle 2 instrument), a micro-Raman spectrometer, and a bespoke (custom-designed) X-ray diffractometer. All experiments were conducted within the engineering constraints of in situ operations to generate realistic data and address the practical challenges of measurement. Our results demonstrate the importance of an integrated approach for this type of work. Each technique made a proportionate contribution to the overall effectiveness of our "pseudopayload" for biogenic assessment of samples yet highlighted a number of limitations of current space instrument technology for in situ astrobiology.

Raman spectroscopy of natural accumulated paraffins from rocks: evenkite, ozokerite and hatchetine.

Raman spectra were obtained from the natural crystalline (evenkite) and amorphous (ozokerite and hatchetin) hydrocarbons, originating from volcanic and sedimentary rocks from several sites (Merník, Evenki, Zastávka). Raman spectra of all materials investigated confirm their exclusively aliphatic character. Vibrational assignments are proposed and compared with pure synthetic compounds and the differences in Raman spectra obtained from materials from different sites are discussed. Monoclinic evenkite corresponds to n-tetratracosane (C(24)H(50)). Noncrystalline waxy ozokerite and hatchetine contain several degradation products in addition to higher paraffins.

Raman microspectroscopic studies of amber resins with insect inclusions.

Raman microscope spectra of specimens of Baltic and Mexican amber resins containing insect inclusions have been analysed using near-infrared excitation to assess the potential for discrimination between the keratotic remains of the insects and the terpenoid matrix. For the Mexican amber specimen the insect spectra exhibit evidence of significant protein degradation compared with the insect remains in the Baltic amber specimen. In both cases the Raman spectra of the insect remains are still distinguishable from the amber resins. Despite its better preservation, however, no spectra could be obtained from the inside of the larger insect preserved in the Baltic amber in agreement with the observation that most insect inclusions in amber are hollow. It is noted that the Mexican amber insect is located adjacent to a large gas bubble in the amber matrix, to which the observed degradation of the insect and its poor state of preservation are attributed. It is concluded that Raman spectra of insect inclusions can provide useful information about the chemical composition of the remains and that confocal microscopy is particularly advantageous in this respect.

Combined FT-Raman spectroscopic and mass spectrometric study of ancient Egyptian sarcophagal fragments.

The application of combined Raman spectroscopic and GC-MS analytical techniques for the characterisation of organic varnish residues from Egyptian Dynastic funerary sarcophagal and cartonnage fragments from the Graeco-Roman period, ca. 2200 BP, is described. The nondestructive use of Raman spectroscopy was initially employed to derive information about the specific location of organic material on the specimens, which were then targeted in specific areas using minimal sampling for GC-MS analysis. In the case of the sarcophagal fragment, a degraded yellow-brown surface treatment was identified as a Pistacia spp. resin; this provides additional evidence for the use of this resin, which has previously been identified in Canaanite transport amphorae, varnishes and "incense" bowls in an Egyptian Late Bronze Age archaeological context. The cartonnage fragment also contained an organic coating for which the Raman spectrum indicated a degradation that was too severe to facilitate identification, but the GC-MS data revealed that it was composed of a complex mixture of fatty acid residues. The combined use of GC-MS and Raman spectroscopy for the characterisation of organic materials in an archaeological context is advocated for minimisation of sampling and restriction to specifically identified targets for museum archival specimens.

FT-Raman spectroscopic study of calcium-rich and magnesium-rich carbonate minerals.

Calcium and magnesium carbonates are important minerals found in sedimentary environments. Although sandstones are the most common rock colonized by endolith organisms, the production of calcium and magnesium carbonates is important in survival strategies of organisms and as a source for the removal of oxalate ions. Extremophile organisms in some situations may convert or destroy carbonates of calcium and magnesium, which gives important information about the conditions under which these organisms can survive. The identification on the surface of Mars of 'White Rock' formations, in Juventae Chasma or Sabaea Terra, as possibly carbonate rocks makes the study of these minerals a prerequisite of remote Martian exploration. Here, we show the protocol for the identification by Raman spectroscopy of different calcium and magnesium carbonates and we present a database of relevance in the search for life, extinct or extant, on Mars; this will be useful for the assessment of data obtained from remote, miniaturized Raman spectrometers now proposed for Mars exploration.

Raman spectroscopic analysis of cyanobacterial gypsum halotrophs and relevance for sulfate deposits on Mars.

The Raman spectra of cyanobacterial species, Gloecapsa and Nostoc, in clear gypsum crystals from the Haughton Crater, Devon Island, Canadian High Arctic, site of a meteorite impact during the Miocene some 23 Mya, have been recorded using several visible and near-infrared excitation wavelengths. The best spectra were obtained using a green wavelength at 514.5 nm and a confocal microscope with an image footprint of about 2 micro in diameter and 2 micro theoretical depth. Raman biosignatures for beta-carotene and scytonemin were obtained for one type of colony and parietin and beta-carotene for another; chlorophyll was detected in both types of colony. The different combination of these radiation protectant biomolecules suggests that the two cyanobacterial colonies, namely Nostoc and Gloecapsa, are adopting different survival strategies in the system. Confocal spectroscopic probing of the gypsum crystals exhibited sufficient discrimination for the identification of the biomolecules through the gypsum crystal, in simulation of the detection of extant or extinct halotrophs. This supports the viability of Raman spectroscopic techniques for incorporation as part of the instrumentation suite of a robotic lander for planetary surface exploration for the detection of organisms inside sulfate crystals from previous hydrothermal activity on Mars.

Raman spectroscopy of hot desert, high altitude epilithic lichens.

Twenty-three highly-coloured lichen specimens belonging to the genera Candelariella, Aspicilia and Xanthoria from high altitude sites in the Atacama Desert, Chile, 2300-4500 metres, have been analysed non-destructively by Raman spectroscopy. The vibrational band assignments in the spectra of the specimens, which were still attached to their limestone or sandstone substrata, were accomplished by comparison with the chemical compositions obtained from wet chemical extraction methods. Carotenoids and chlorophyll were found in all specimens as major components and the characteristic spectral signatures of calcium oxalate monohydrate (whewellite) and dihydrate (weddellite) could be identified; chemical signatures were found for these materials even in lichen thalli growing on the non-calcareous substrata, indicating probably that the calcium was provided here from wind- or rain-borne sources. The Raman spectral biomarkers for a variety of protective biomolecules and accessory pigments such as usnic acid, calycin, pulvinic acid dilactone and rhizocarpic acid have been identified in the lichens, in broad agreement with the chemical extraction profiles. The present study indicates that some form of non-destructive taxonomic identification based on Raman spectroscopy was also possible.

Raman spectroscopy of endoliths from Antarctic cold desert environments.

Six endolithic communities from Antarctic cold desert environments have been analysed by Raman spectroscopy. The extreme conditions that the organisms have to withstand in cold environments leads to the adoption of different survival strategies and adaptation of the geological environment. Production of radiation- and desiccation-protective biomolecules is identifiable but the displacement of potentially protective minerals onto the rock surface has also been detected as a protective mechanism against UV-radiation. In this work, Raman spectroscopy is demonstrated as a valuable technique to determine the organic and inorganic compounds used by microorganisms as protective mechanisms against extreme stress conditions. The data from this study will be useful for construction of molecular recognition biomarkers and remote Raman spectral sensing experiments proposed for terrestrial extremophiles in stressed environments.

Lichen biodeterioration of ecclesiastical monuments in northern Spain.

Seven highly-coloured lichen species belonging to the genera Caloplaca, Candelariella, Aspicilia and Xanthoria from ecclesiastical buildings in northern Spain have been analysed non-destructively by FT-Raman spectroscopy. The vibrational band assignments in the spectra of the specimens, which were still attached to their limestone or sandstone substrata, were accomplished with the assistance of the chemical compositions obtained from wet chemical extraction methods. beta-Carotene was found in all specimens as the major pigment, and the characteristic spectral signatures of calcium oxalate monohydrate (whewellite) and dihydrate (weddelite) could be identified; chemical signatures were found for these materials even in lichen thalli growing the non-calcareous substrata, indicating, probably, that the calcium was provided here from wind-or-rain-borne sources. The Raman spectral biomarkers found in the lichens broadly agreed with the chemical extraction profiles as expected, but the present study indicates that some form of non-destructive taxonomic identification based on Raman spectroscopy was possible.