Petr Vítek

Evaluation of portable Raman spectrometer with 1064 nm excitation for geological and forensic applications.

The development of miniaturized Raman instrumentation is in demand for applications relevant to forensic, pharmaceutical and art analyses, as well as geosciences, and planetary exploration. In this study we report on evaluation of a portable dispersive Raman spectrometer equipped with 1064 nm laser excitation. Selected samples from geological, geobiological and forensic areas of interest have been studied from which the advantages, disadvantages and the analytical potential of the instrument are assessed based on a comparison with bench instrumentation and other portable Raman spectrometers using 785 nm excitation. It is demonstrated that the instrument operating with 1064 nm excitation has potential for expanding the number and types of samples that can be measured by miniaturized Raman spectroscopy without interfering fluorescence background emission. It includes inorganic and organic minerals, biomolecules within living lichen and endolithic cyanobacteria as well as drugs of abuse and explosives.

Application of portable Raman instruments for fast and non-destructive detection of minerals on outcrops

Raman spectral signatures have been obtained in situ for a series of minerals using portable Raman instruments. Cerussite, anglesite, wulfenite, titanite, calcite, tremolite, andradite and quartz were detected using portable Raman spectrometer First Defender XL (Ahura). Baryte, almandine and realgar Raman spectra obtained by this instrument in the field were compared to the data measured by the other mobile Raman instrument Inspector Raman (DeltaNu). Bench Raman dispersive microspectrometer (InVia Reflex, Renishaw) was used for comparative purposes. All spectra were obtained using a 785nm diode excitation. Although displaying lower spectral resolution comparing with the laboratory confocal instrument both portable instruments permit unambiguous detection of minerals in the field. These possibilities designate portable Raman machines as excellent tools for field geological applications. Miniaturised Raman instrument combined with LIBS will be included in the payload of the EXO Mars mission and would open interesting research possibilities in other in situ field planetary studies.

Microbial colonization of halite from the hyper-arid Atacama Desert studied by Raman spectroscopy

The hyper-arid core of the Atacama Desert (Chile) is the driest place on Earth and is considered a close analogue to the extremely arid conditions on the surface of Mars. Microbial life is very rare in soils of this hyper-arid region, and autotrophic micro-organisms are virtually absent. Instead, photosynthetic micro-organisms have successfully colonized the interior of halite crusts, which are widespread in the Atacama Desert. These endoevaporitic colonies are an example of life that has adapted to the extreme dryness by colonizing the interior of rocks that provide enhanced moisture conditions. As such, these colonies represent a novel example of potential life on Mars. Here, we present non-destructive Raman spectroscopical identification of these colonies and their organic remnants. Spectral signatures revealed the presence of UV-protective biomolecules as well as light-harvesting pigments pointing to photosynthetic activity. Compounds of biogenic origin identified within these rocks differed depending on the origins of specimens from particular areas in the desert, with differing environmental conditions. Our results also demonstrate the capability of Raman spectroscopy to identify biomarkers within rocks that have a strong astrobiological potential.

The miniaturized Raman system and detection of traces of life in halite from the Atacama Desert: some considerations for the search for life signatures on Mars.

Raman spectroscopy is being adopted as a nondestructive instrumentation for the robotic exploration of Mars to search for traces of life in the geological record. Here, miniaturized Raman spectrometers of two different types equipped with 532 and 785 nm lasers for excitation, respectively, were compared for the detection of microbial biomarkers in natural halite from the hyperarid region of the Atacama Desert. Measurements were performed directly on the rock as well as on the homogenized, powdered samples prepared from this material-the effects of this sample preparation and the excitation wavelength employed in the analysis are compared and discussed. From these results, 532 nm excitation was found to be superior for the analysis of powdered specimens due to its high sensitivity toward carotenoids and hence a higher capability for their detection at relatively low concentration in bulk powdered specimens. For the same reason, this wavelength was a better choice for the detection of carotenoids in direct measurements made on the rock samples. The 785 nm excitation wavelength, in contrast, proved to be more sensitive toward the detection of scytonemin.

Raman spectroscopic analysis of geological and biogeological specimens of relevance to the ExoMars mission.

A novel miniaturized Raman spectrometer is scheduled to fly as part of the analytical instrumentation package on an ESA remote robotic lander in the ESA/Roscosmos ExoMars mission to search for evidence for extant or extinct life on Mars in 2018. The Raman spectrometer will be part of the first-pass analytical stage of the sampling procedure, following detailed surface examination by the PanCam scanning camera unit on the ExoMars rover vehicle. The requirements of the analytical protocol are stringent and critical; this study represents a laboratory blind interrogation of specimens that form a list of materials that are of relevance to martian exploration and at this stage simulates a test of current laboratory instrumentation to highlight the Raman technique strengths and possible weaknesses that may be encountered in practice on the martian surface and from which future studies could be formulated. In this preliminary exercise, some 10 samples that are considered terrestrial representatives of the mineralogy and possible biogeologically modified structures that may be identified on Mars have been examined with Raman spectroscopy, and conclusions have been drawn about the viability of the unambiguous spectral identification of biomolecular life signatures. It is concluded that the Raman spectroscopic technique does indeed demonstrate the capability to identify biomolecular signatures and the mineralogy in real-world terrestrial samples with a very high degree of success without any preconception being made about their origin and classification.

Phototrophic Community in Gypsum Crust from the Atacama Desert Studied by Raman Spectroscopy and Microscopic Imaging

The hyperarid core of the Atacama Desert represents one of the driest places on Earth with an exceptional occurrence of microbial life coping with extreme environmental stress factors. The gypsum crusts have already been found to harbor diverse communities in this area. Here, we present a Raman spectroscopic study, complemented by correlative microscopic imaging using SEM-BSE and fluorescence microscopy, of the endolithic microbial communities inside the Ca-sulphate crusts dominated by phototrophic microorganisms. Differences of pigment composition within different zones follow the cyanobacterial and algal colonization and also reveal the degradation of phycobiliproteins within the decayed biomass of cyanobacteria. Carotenoids of at least three different types were recognized, differing in dependence on the particular phylum of phototrophic microorganisms. Moreover, calcium oxalate monohydrate – whewellite – was found to be associated with the algae and hyphal associations living in the lower regions of the crust. The 785 nm excitation wavelength employed here was found to be the correct source for studying pigment composition as well as for the detection of the oxalate. A comparison of these results with those using 514.5 nm laser excitation which is widely adopted for the detection of carotenoids due to the resonance Raman effect is made and discussed.

Practical Considerations for the Field Application of Miniaturized Portable Raman Instrumentation for the Identification of Minerals

The nondestructive identification of both inorganic and organic compounds without the need for chemical or mechanical sample preparation is an advantage of the Raman spectroscopic analytical technique when applied in situ using miniaturized equipment for the geosciences. This is critically assessed here for several real life geoscientific scenarios in which several groups of minerals were analyzed with emphasis on evaporites, carbonates, and selected types of dark minerals and weak Raman scatterers. The role of individual analytical instrumental parameters such as focal plane precision, exposure time, and ambient light conditions that can affect the acquisition and interpretation of spectroscopic data from these specimens in field conditions was also evaluated.

Adaptation strategies of endolithic chlorophototrophs to survive the hyperarid and extreme solar radiation environment of the Atacama Desert

The Atacama Desert, northern Chile, is one of the driest deserts on Earth and, as such, a natural laboratory to explore the limits of life and the strategies evolved by microorganisms to adapt to extreme environments. Here we report the exceptional adaptation strategies of chlorophototrophic and eukaryotic algae, and chlorophototrophic and prokaryotic cyanobacteria to the hyperarid and extremely high solar radiation conditions occurring in this desert. Our approach combined several microscopy techniques, spectroscopic analytical methods, and molecular analyses. We found that the major adaptation strategy was to avoid the extreme environmental conditions by colonizing cryptoendolithic, as well as, hypoendolithic habitats within gypsum deposits. The cryptoendolithic colonization occurred a few millimeters beneath the gypsum surface and showed a succession of organized horizons of algae and cyanobacteria, which has never been reported for endolithic microbial communities. The presence of cyanobacteria beneath the algal layer, in close contact with sepiolite inclusions, and their hypoendolithic colonization suggest that occasional liquid water might persist within these sub-microhabitats. We also identified the presence of abundant carotenoids in the upper cryptoendolithic algal habitat and scytonemin in the cyanobacteria hypoendolithic habitat. This study illustrates that successful lithobiontic microbial colonization at the limit for microbial life is the result of a combination of adaptive strategies to avoid excess solar irradiance and extreme evapotranspiration rates, taking advantage of the complex structural and mineralogical characteristics of gypsum deposits—conceptually called “rocks habitable architecture.” Additionally, self-protection by synthesis and accumulation of secondary metabolites likely produces a shielding effect that prevents photoinhibition and lethal photooxidative damage to the chlorophototrophs, representing another level of adaptation.

Raman spectroscopy of the Dukhan sabkha: identification of geological and biogeological molecules in an extreme environment

The characterization of minerals and biogeological deposits in a terrestrial Arabian sabkha has a direct relevance for the exploration of Mars since the discovery by the NASA rovers Spirit and Opportunity of evaporate minerals on Mars that could have arisen from aquifers and subsurface water movement. The recognition of carbonates and sulphates in Gusev Crater has afforded an additional impetus to these studies, as relict or extant microbial extremophilic organisms could have colonized these geological matrices, as has been recorded on Earth. Here, we describe the Raman spectroscopic analysis of specimens of evaporitic materials sampled from the Dukhan sabkha, the largest inland sabkha in the Persian Gulf. With daily temperatures reaching in excess of 60°C and extreme salinity, we have identified the characteristic Raman signatures of key biomolecular compounds in association with evaporitic minerals and geological carbonate and sulphate matrices, which indicate that extremophilic cyanobacterial colonies are existent there. This evidence, the first to be acquired spectroscopically from such a region, establishes a platform for further studies using remote, portable Raman instrumentation that will inform the potential of detection of similar systems on the Martian surface or subsurface in future space missions. A comparison is made between the results from this study and the previous analysis of a gypsum/halite sabkha where the extremophilic molecular signatures were better preserved.

Use of Raman spectroscopy for identification of compatible solutes in halophilic bacteria.

We explored the use of Raman spectroscopy to detect organic osmotic solutes as biomarkers in the moderately halophilic heterotrophic bacterium Halomonas elongata grown in complex medium (accumulation of glycine betaine) and in defined medium with glucose as carbon source (biosynthesis of ectoine), and in the anoxygenic phototrophic Ectothiorhodospiramarismortui known to synthesize glycine betaine in combination with minor amounts of trehalose and N-α-carbamoyl glutamineamide. We tested different methods of preparation of the material: lyophilization, two-phase extraction of water-soluble molecules, and perchlorate extraction. Raman signals of glycine betaine and ectoine were detected; perchlorate extraction followed by desalting the extract on an ion retardation column gave the best results. Lyophilized cells of E.marismortui showed strong signals of carotenoid pigments, and glycine betaine could be detected only after perchlorate extraction and desalting. The data presented show that Raman spectroscopy is a suitable tool to assess the mode of osmotic adaptation used by halophilic microorganisms.