Assessment of Raman spectroscopy as a tool for the non-destructive identification of organic minerals and biomolecules for Mars studies

Several characteristic geological features found on the surface of Mars by planetary rovers suggest that a possible extinct biosphere could exist based on similar sources of energy as occurred on Earth. For this reason, analytical instrumental protocols for the detection of biomarkers in suitable geological matrices unequivocally have to be elaborated for future unmanned explorations including the forthcoming ESA ExoMars mission. As part of the Pasteur suite of analytical instrumentation on ExoMars, the Raman/LIBS instrument will seek elemental and molecular information about geological, biological and biogeological markers in the Martian record. A key series of experiments on terrestrial Mars analogues, of which this paper addresses a particularly important series of compounds, is required to obtain the Raman spectra of key molecules and crystals, which are characteristic for each biomarker. Here, we present Raman spectra of several examples of organic compounds which have been recorded non-destructively—higher n-alkanes, polycyclic aromatic hydrocarbons, carotenoids, salts of organic acids, pure crystalline terpenes as well as oxygen-containing organic compounds. In addition, the lower limit of β-carotene detection in sulphate matrices using Raman microspectroscopy was estimated.     

Volcaniclastic habitats for early life on Earth and Mars: A case study from ∼3.5 Ga-old rocks from the Pilbara, Australia

Within the context of present and future in situ missions to Mars to investigate its habitability and to search for traces of life, we studied the habitability and traces of past life in ∼3.5 Ga-old volcanic sands deposited in littoral environments an analogue to Noachian environments on Mars. The environmental conditions on Noachian Mars (4.1-3.7 Ga) and the Early Archaean (4.0-3.3 Ga) Earth were, in many respects, similar: presence of liquid water, dense CO₂ atmosphere, availability of carbon and bio-essential elements, and availability of energy. For this reason, information contained in Early Archaean terrestrial rocks concerning habitable conditions (on a microbial scale) and traces of past life are of relevance in defining strategies to be used to identify past habitats and past life on Mars.One such example is the 3.446 Ga-old Kitty’s Gap Chert in the Pilbara Craton, NW. Australia. This formation consists of volcanic sediments deposited in a coastal mudflat environment and is thus a relevant analogue for sediments deposited in shallow water environments on Noachian Mars. Two main types of habitat are represented, a volcanic (lithic) habitat and planar stabilized sediment surfaces in sunlit shallow waters. The sediments hosted small (<1 μm in size) microorganisms that formed colonies on volcanic particle surfaces and in pore waters within the volcanic sediments, as well as biofilms on stabilised sediment surfaces. The microorganisms included coccoids, filaments and rare rod-shaped organisms associated with microbial polymer (EPS). The preserved microbial community was apparently dominated by chemotrophic organisms but some locally transported filaments and filamentous mat fragments indicate that possibly photosynthetic mats formed nearby. Both microorganisms and sediments were silicified during very early diagenesis.There are no macroscopic traces of fossilised life in these volcanic sediments and sophisticated instrumentation and specialized sample preparation techniques are required to establish the biogenicity and syngenicity of the traces of past life. The fact that the traces of life are cryptic, and the necessity of using sophisticated instrumentation, reinforces the challenges and difficulties of in situ robotic missions to identify past life on Mars. We therefore recommend the return of samples from Mars to Earth for a definitive search for traces of life.     

A search for a nonbiological explanation of the Viking Labeled Release life detection experiment

The Viking Labeled Release (LR) data obtained on Mars satisfy the criteria established for a biological response. The importance of the issue, especially when viewed against the harsh environment on Mars, requires careful consideration of possible nonbiological reactions that may have produced false positive results. A $\text{3}\text{1}\text{2}\text{-}\text{year}$ laboratory effort to investigate possible chemical, physical, and physicochemical agents or mechanisms has been concluded. Among nonbiological possibilities, hydrogen peroxide, putatively on Mars, emerged as the principal candidate. When placed on analog Mars soils prepared to match the Viking inorganic analysis of the Mars surface material, hydrogen peroxide did not duplicate the LR Mars data. When other materials were used as substrate, hydrogen peroxide could be made to evoke the type of responses obtained by the LR Mars experiment. However, essential criteria concerning the formation, accuumulation, and preservation of hydrogen peroxide to qualify it as the active agent on Mars have not been met and new data show it to be essentially absent from the Mars atmosphere. The presence of a biological agent on Mars must still be considered. This interpretation of the LR results is strengthened by a recent report that the Viking organic analysis instrument (GCMS) failed to detect organics in an Antarctic soil in which the LR instrument had demonstrated the presence of microorganisms.     

The miniaturized Möessbauer spectrometer MIMOS II for the Phobos-Grunt mission

Möessbauer spectroscopy is a powerful tool for the mineralogical analysis of Fe-bearing materials. The miniaturized Möessbauer spectrometer MIMOS II has already been working on the surface of Mars for 6 years as part of the NASA Mars Exploration Rovers mission. The improved version of the instrument is a component of the scientific payload of the Phobos-Grunt mission. The scientific objectives of the instrument are the following: to identify the iron-bearing phases, to determine the quantitative distribution of iron among these phases, and to determine the distribution of iron among its oxidation states.     

The laser ablation ion funnel: Sampling for in situ mass spectrometry on Mars

A considerable investment has been made by NASA and other space agencies to develop instrumentation suitable for in situ analytical investigation of extra terrestrial bodies including various mass spectrometers (time-of-flight, quadrupole ion trap, quadrupole mass filters, etc.). However, the front-end sample handling that is needed to collect and prepare samples for interrogation by such instrumentation remains underdeveloped. Here we describe a novel approach tailored to the exploration of Mars where ions are created in the ambient atmosphere via laser ablation and then efficiently transported into a mass spectrometer for in situ analysis using an electrodynamic ion funnel. This concept would enable elemental and isotopic analysis of geological samples with the analysis of desorbed organic material a possibility as well. Such an instrument would be suitable for inclusion on all potential missions currently being considered such as the Mid-Range Rover, the Astrobiology Field Laboratory, and Mars Sample Return (i.e., as a sample pre-selection triage instrument), among others.     

The specific molecular identification of life experiment (SMILE)

We describe a compact, highly integrated instrument concept for the detection and identification of a wide range of molecules associated with extinct/extant life or potential life processes. The Specific Molecular Identification of Life Experiment (SMILE) will be sensitive to the presence of a range of target molecules using both electrical and optical transduction techniques, and incorporates molecular imprinted polymers in addition to traditional biological receptors such as antibodies. A number of versions of the concept are possible depending on available resources e.g. mass, volume, etc. The full concept utilises a novel imaging interferometer where a large number of molecular receptors are deposited on the measurement plane of an imaging interferometer and read out by an imaging detector, enabling multiple targets—biomarkers—within a sample to be measured simultaneously. The optics can also form the basis of an UV-NIR imaging Fourier spectrometer allowing basic mineralogy studies to be conducted using optical properties to assist in the determination of the geological context of the samples. By incorporating micro-fabricated transducer arrays, micro-fluidics and artificial molecular recognition systems, as well as recombinant antibody technology with appropriate integration methods, SMILE forms a compact and robust “Life Marker Chip” which has been proposed for future planetary missions including ESA's ExoMars mission, where the instrument offers the possibility of conducting a direct in situ search for signs of past or present biological activity on Mars. In addition to its role in planetary exploration, derivatives of SMILE have multiple terrestrial applications in fields such as forensic analysis and environmental monitoring.     

In situ methods for measuring thermal properties and heat flux on planetary bodies

The thermo-mechanical properties of planetary surface and subsurface layers control to a high extent in which way a body interacts with its environment, in particular how it responds to solar irradiation and how it interacts with a potentially existing atmosphere. Furthermore, if the natural temperature profile over a certain depth can be measured in situ, this gives important information about the heat flux from the interior and thus about the thermal evolution of the body. Therefore, in most of the recent and planned planetary lander missions experiment packages for determining thermo-mechanical properties are part of the payload. Examples are the experiment MUPUS on Rosetta's comet lander Philae, the TECP instrument aboard NASA's Mars polar lander Phoenix, and the mole-type instrument HP³ currently developed for use on upcoming lunar and Mars missions. In this review we describe several methods applied for measuring thermal conductivity and heat flux and discuss the particular difficulties faced when these properties have to be measured in a low pressure and low temperature environment. We point out the abilities and disadvantages of the different instruments and outline the evaluation procedures necessary to extract reliable thermal conductivity and heat flux data from in situ measurements.     

Extended survival of several organisms and amino acids under simulated martian surface conditions

Recent orbital and landed missions have provided substantial evidence for ancient liquid water on the martian surface as well as evidence of more recent sedimentary deposits formed by water and/or ice. These observations raise serious questions regarding an independent origin and evolution of life on Mars. Future missions seek to identify signs of extinct martian biota in the form of biomarkers or morphological characteristics, but the inherent danger of spacecraft-borne terrestrial life makes the possibility of forward contamination a serious threat not only to the life detection experiments, but also to any extant martian ecosystem. A variety of cold and desiccation-tolerant organisms were exposed to 40 days of simulated martian surface conditions while embedded within several centimeters of regolith simulant in order to ascertain the plausibility of such organisms’ survival as a function of environmental parameters and burial depth. Relevant amino acid biomarkers associated with terrestrial life were also analyzed in order to understand the feasibility of detecting chemical evidence for previous biological activity. Results indicate that stresses due to desiccation and oxidation were the primary deterrent to organism survival, and that the effects of UV-associated damage, diurnal temperature variations, and reactive atmospheric species were minimal. Organisms with resistance to desiccation and radiation environments showed increased levels of survival after the experiment compared to organisms characterized as psychrotolerant. Amino acid analysis indicated the presence of an oxidation mechanism that migrated downward through the samples during the course of the experiment and likely represents the formation of various oxidizing species at mineral surfaces as water vapor diffused through the regolith. Current sterilization protocols may specifically select for organisms best adapted to survival at the martian surface, namely species that show tolerance to radical-induced oxidative damage and low water activity environments. Additionally, any hypothetical martian ecosystems may have evolved similar physiological traits that allow sporadic metabolism during periods of increased water activity.     

A search for amino acids and nucleobases in the Martian meteorite Roberts Massif 04262 using liquid chromatography‐mass spectrometry

The investigation into whether Mars contains signatures of past or present life is of great interest to science and society. Amino acids and nucleobases are compounds that are essential for all known life on Earth and are excellent target molecules in the search for potential Martian biomarkers or prebiotic chemistry. Martian meteorites represent the only samples from Mars that can be studied directly in the laboratory on Earth. Here, we analyzed the amino acid and nucleobase content of the shergottite Roberts Massif (RBT) 04262 using liquid chromatography‐mass spectrometry. We did not detect any nucleobases above our detection limit in formic acid extracts; however, we did measure a suite of protein and nonprotein amino acids in hot‐water extracts with high relative abundances of β‐alanine and γ‐amino‐n‐butyric acid. The presence of only low (to absent) levels of several proteinogenic amino acids and a lack of nucleobases suggest that this meteorite fragment is fairly uncontaminated with respect to these common biological compounds. The distribution of straight‐chained amine‐terminal n‐ω‐amino acids in RBT 04262 resembled those previously measured in thermally altered carbonaceous meteorites (Burton et al. 2012; Chan et al. 2012). A carbon isotope ratio of ?24‰ ± 6‰ for β‐alanine in RBT 04262 is in the range of reduced organic carbon previously measured in Martian meteorites (Steele et al. 2012). The presence of n‐ω‐amino acids may be due to a high temperature Fischer‐Tropsch‐type synthesis during igneous processing on Mars or impact ejection of the meteorites from Mars, but more experimental data are needed to support these hypotheses.     

A spectroscopic method for identifying terrestrial biocarbonates and application to Mars

Searching for traces of extinct and/or extant life on Mars is one of the major objectives for remote-sensing and in situ exploration of the planet. In previous laboratory works we have investigated the infrared spectral modifications induced by thermal processing on different carbonate samples, in the form of fresh shells and fossils of different ages, whose biotic origin is easily recognizable. The goal was to discriminate them from their abiotic counterparts. In general, it is difficult to identify biotic signatures, especially when the organisms inducing the carbonate precipitation have low fossilization potential (i.e. microbes, bacteria, archaea). A wide variety of microorganisms are implicated in carbonate genesis, and their direct characterization is very difficult to evaluate by traditional methods, both in ancient sedimentary systems and even in recent environments.In the present work we apply our analysis to problematic carbonate samples, in which there is no clear evidence of controlled or induced biomineralization. This analysis indicates a very likely biotic origin of the aragonite samples under study, in agreement with the conclusion previously reported by Guido et al. (2007) who followed a completely different approach based on a complex set of sedimentary, petrographic, geochemical and biochemical analyses. We show that our method is reliable for discriminating between biotic and abiotic carbonates, and therefore it is a powerful tool in the search for life on Mars in the next generation of space missions to the planet.     

Spectral reflectance properties of carbonates from terrestrial analogue environments: Implications for Mars

Spectroscopic analysis of carbonate-bearing samples from a variety of terrestrial environments provides important insights into spectroscopy-based investigations of Mars designed to detect the presence of carbonate minerals. In order to better address the spectral detectability of carbonates on Mars, we examined the spectral reflectance properties of carbonates and carbonate-bearing lithologies from a variety of terrestrial environments, including impact structures (Haughton, St. Martin, Eagle Butte), landslides (Frank), quarrying operations (Hecla), carbonates affected by weathering (Haughton, East German Creek), and sulfide-sulfate-carbonate assemblages (Central Manitoba). The goal is to identify processes and environments that can affect spectroscopy-based carbonate detection, for more detailed follow-on studies. Common carbonates appear to be stable, from a spectroscopic perspective, to various tectonic processes. Iron oxides/hydroxides do not appear to significantly affect spectral detectability of carbonates, as the spectrum-altering effects of these phases are largely restricted to the region below ∼1 μm, while useful carbonate absorption bands occur longward of ∼1.8 μm. Carbonate detection and characterization in the 0.35-2.5-μm region is largely restricted to a single absorption feature in the 2.3-μm region, which can be problematic for robust carbonate identification. While tectonic processes and iron oxide/hydroxide staining do not appear to significantly impair carbonate detection based on the 2.3-μm region absorption band, a number of other factors can affect carbonate detection. These include the fact that this absorption band is weak compared to many other minerals, a number of other minerals also exhibit absorption bands in this wavelength region (leading to possible misidentifications), and that even small abundances of minerals that absorb strongly in this region will reduce the strength of the carbonate absorption band. Identifying the nature of accessory minerals associated with carbonates can be used to constrain possible formation environments. Ongoing research at carbonate-bearing terrestrial analogue sites will continue to provide new insights into the occurrence and detection of carbonates on Mars.     

Raman spectroscopic analysis of cyanobacterial colonization of hydromagnesite, a putative martian extremophile

Raman spectra of an extremophile cyanobacterial colony in hydromagnesite from Lake Salda in Turkey have revealed a biogeological modification which is manifest as aragonite in the stratum associated with the colony. The presence of key spectral biomarkers of organic protectant molecules such as β-carotene and scytonemin indicate that the survival strategy of the cyanobacteria is significantly one of UV-radiation protection. The terrestrial location of this extremophile is worthy of consideration further because of its possible putative link with the “White Rock” formations in Sabaea Terra and Juventae Chasma on Mars.     

High spectral resolution UV to near-IR observations of Mars using HST/STIS

We acquired high spectral and spatial resolution hyperspectral imaging spectrometer observations of Mars from near-UV to near-IR wavelengths (∼300 to 1020 nm) using the STIS instrument on the Hubble Space Telescope during the 1999, 2001, and 2003 oppositions. The data sets have been calibrated to radiance factor (I/F) and map-projected for comparison to each other and to other Mars remote sensing measurements. We searched for and (where detected) mapped a variety of iron-bearing mineral signatures within the data. The strong and smooth increase in I/F from the near-UV to the visible that gives Mars its distinctive reddish color indicates that poorly crystalline ferric oxides dominate the spectral properties of the high albedo regions (as well as many intermediate and low albedo regions), a result consistent with previous remote sensing studies of Mars at these wavelengths. In the near-IR, low albedo regions with a negative spectral slope and/or a distinctive ∼900 nm absorption feature are consistent with, but not unique indicators of, the presence of high-Ca pyroxene or possibly olivine. Mixed ferric-ferrous minerals could also be responsible for the ∼900 nm feature, especially in higher albedo regions with a stronger visible spectral slope. We searched for the presence of several known diagnostic absorption features from the hydrated ferric sulfate mineral jarosite, but did not find any unique evidence for its occurrence at the spatial scale of our observations. We identified a UV contrast reversal in some dark region spectra: at wavelengths shorter than about 340 nm these regions are actually brighter than classical bright regions. This contrast reversal may be indicative of extremely “clean” low albedo surfaces having very little ferric dust contamination. Ratios between the same regions observed during the planet-encircling dust storm of 2001 and during much clearer atmospheric conditions in 2003 provide a good direct estimate of the UV to visible spectral characteristics of airborne dust aerosols. These HST observations can help support the calibration of current and future Mars orbital UV to near-IR spectrometers, and they also provide a dramatic demonstration that even at the highest spatial resolution possible to achieve from the Earth, spectral variations on Mars at these wavelengths are subtle at best.     

The electrical properties of Mars analogue dust

Dust is a major environmental factor on the surface and in the atmosphere of Mars. Knowing the electrical charge state of this dust would be of both scientific interest and important for the safety of instruments on the Martian surface. In this study the first measurements have been performed of dust electrification using suspended Mars analogue material. This has been achieved by attracting suspended dust onto electrodes placed inside a Mars simulation wind tunnel. The Mars analogue used was from Salten Skov in Denmark, this contained a high concentration of ferric oxide precipitate. Once suspended, this dust was found to consist of almost equal quantities of negatively (46±6%) and positively (44±15%) charged grains.These grains were estimated to typically carry a net charge of around 10⁵e, this is sufficient to dominate the processes of adhesion and cohesion of this suspended dust. Evidence is presented for electrostatic aggregation of the dust while in suspension. Development of a simple instrument for measuring electrical charging of the suspended dust on Mars will be discussed.     

Stereoscopic observations of solar flares made onboard the 2001 Mars Odyssey spacecraft and CORONAS-F satellite

The results of observations of solar hard radiation recorded by two spacecraft—2001 Mars Odyssey and CORONAS-F—which were located in the vicinity of Mars and Earth, respectively, are discussed. The HEND instrument, developed at the Space Research Institute of the Russian Academy of Sciences, recorded photons with energies ranging from 80 keV to 2 MeV, and the SPR and SONG instruments, developed at the Skobeltsyn Research Institute of Nuclear Physics of the Moscow State University, detected radiation in the energy interval from 15 keV to 100 MeV. The rising of the sunspot group 10486 in late October 2003, which had been observed from Martian orbit before it was seen from the Earth’s surface, is analyzed in detail. In this case, observations made from directions that differ by 24° showed a close-to-24 h advance for the detection of hard radiation of flares. Stereoscopic observations of M-class flares near the limb show that the overwhelming part of radiation with energies above 80 keV arises at heights that do not exceed 7–10 thousand km. Also reported are the results of observations of the powerful flare on August 25, 2001, by the two devices, which complement each other substantially. The processes resulting in the formation of high-energy radiation of solar flares are discussed.     

On the properties of O and O2 ions in a hybrid model and in Mars Express IMA/ASPERA-3 data: A case study

We have performed a numerical simulation to analyze the energy spectra of escaping planetary O⁺ and O₂⁺ ions at Mars. The simulated time-energy spectrograms were generated along orbit no. 555 (June 27, 2004) of Mars Express when its Ion Mass Analyzer (IMA)/ASPERA-3 ion instrument detected escaping planetary ions. The simulated time-energy spectrograms are in general agreement with the hypothesis that planetary O⁺ and O₂⁺ ions far from Mars are accelerated by the convective electric field. The HYB-Mars hybrid model simulation also shows that O⁺ ions originating from the ionized hot oxygen corona result in a high-energy (E>1 keV) O⁺ ion population that exists very close to Mars. In addition, the simulation also results in a low-energy (E<0.1 keV) planetary ion population near the pericenter. In the analyzed orbit, IMA did not observe a clear high-energy planetary ion or a clear low-energy planetary ion population near Mars. One possible source for this discrepancy may be the Martian magnetic crustal anomalies because MEX passed over a strong crustal field region near the pericenter, but the hybrid model does not include the magnetic crustal anomalies.     

Lidar atmospheric measurements on Mars and Earth

The LIDAR instrument operating from the surface of Mars on the Phoenix Mission measured vertical profiles of atmospheric dust and water ice clouds at temperatures around −65 °C. An equivalent lidar system was utilized for measurements in the atmosphere of Earth where dust and cloud conditions are similar to Mars. Coordinated aircraft in situ sampling provided a verification of lidar measurement and analysis methods and also insight for interpretation of lidar derived optical parameters in terms of the dust and cloud microphysical properties. It was found that the vertical distribution of airborne dust above the Australian desert is quite similar to what is observed in the planetary boundary layer above Mars. Comparison with the in situ sampling is used to demonstrate how the lidar derived optical extinction coefficient is related to the dust particle size distribution. The lidar measurement placed a constraint on the model size distribution that has been used for Mars. Airborne lidar measurements were also conducted to study cirrus clouds that form in the Earth’s atmosphere at a similar temperature and humidity as the clouds observed with the lidar on Mars. Comparison with the in situ sampling provides a method to derive the cloud ice water content (IWC) from the Mars lidar measurements.     

Study of terrestrial fossils in phyllosilicate-rich soils: Implication in the search for biosignatures on Mars

The search for biosignatures in the soil of Mars is a major objective of the planet exploration, but the detection of such structures is not straightforward due to the degradation of the organic material. In a previous work our group has analyzed the spectral reaction to thermal processing of biomineral carbonate samples including fresh and fossil shells. We found that two terrestrial fossil shells collected from clay deposits preserve their biomineral characteristics much better than coeval fossils not embedded in clay layers.In the present paper we extend our analysis to a set of fossils found in three different terrestrial clay deposits. Our results confirm that the level of degradation may be much lower than the standard values if fossils are surrounded by clay minerals. As a result these fossils have a spectroscopic response to thermal treatment which make them much more distinguishable from their abiotic counterparts than coeval fossils not collected from clay deposits. This result implies that the phyllosilicates regions recently discovered on Mars may represent very interesting environments that can provide conditions favorable to preserve evidence of biomarkers, and hence can be regarded as good candidate locations for their detection.     

Electromagnetic characterization of polar ice-wedge polygons: Implications for periglacial studies on Mars and Earth

Polygonal terrain is found in a variety of polar environments on Earth and Mars. As a result, many areas of northern Canada may represent ideal terrestrial analogues for specific regions of Mars - in particular the northern plains. In the Canadian Arctic, polygon troughs are commonly underlain by wedges of massive ice, with rare examples of other wedge types. If the same is true for Mars, this raises interesting implications for the processes that concentrate H₂O at the Martian poles. This study uses an electromagnetic induction sensor to investigate the electromagnetic characteristics of terrestrial polar ice-wedge polygons. Surveys were conducted in two regions of the Canadian Arctic using a DUALEM-1S dual-geometry electromagnetic induction sensor, which measures electrical conductivity in the first 1.5-2 m of the subsurface. At locations where strong geomorphological evidence of ice was found, polygon troughs corresponded to local conductive anomalies. Trenching confirmed the presence of ice wedges at one site and allowed ground-truthing and calibration of the geophysical data. Previously unknown bodies of massive ice were also identified through the use of this geophysical technique. This study shows that an electromagnetic induction sounder is a useful instrument for detecting and mapping out the presence of subsurface ice in the Canadian Arctic. Taking together with its small size, portability and ruggedness, we suggest that this would also be a useful instrument for any future missions to Mars’ polar regions.     

Long-term observations of the evolution of the southern seasonal cap of Mars: Neutron measurements by the HEND instrument onboard the 2001 <Emphasis Type="Italic">Mars Odyssey</Emphasis> spacecraft

We present the results of five-year observations of the southern seasonal cap of Mars based on neutron spectroscopy of the surface fulfilled by the Russian HEND instrument onboard the NASA 2001 Mars Odyssey spacecraft. The numerical modeling of the observational data allowed us to reconstruct the curves of the variations of the total mass of the southern seasonal cap of Mars for different years (three Martian years) and to find the year-to-year variations of the seasonal cycle.