SOLID3: a multiplex antibody microarray-based optical sensor instrument for in situ life detection in planetary exploration

The search for unequivocal signs of life on other planetary bodies is one of the major challenges for astrobiology. The failure to detect organic molecules on the surface of Mars by measuring volatile compounds after sample heating, together with the new knowledge of martian soil chemistry, has prompted the astrobiological community to develop new methods and technologies. Based on protein microarray technology, we have designed and built a series of instruments called SOLID (for "Signs Of LIfe Detector") for automatic in situ detection and identification of substances or analytes from liquid and solid samples (soil, sediments, or powder). Here, we present the SOLID3 instrument, which is able to perform both sandwich and competitive immunoassays and consists of two separate functional units: a Sample Preparation Unit (SPU) for 10 different extractions by ultrasonication and a Sample Analysis Unit (SAU) for fluorescent immunoassays. The SAU consists of five different flow cells, with an antibody microarray in each one (2000 spots). It is also equipped with an exclusive optical package and a charge-coupled device (CCD) for fluorescent detection. We demonstrated the performance of SOLID3 in the detection of a broad range of molecular-sized compounds, which range from peptides and proteins to whole cells and spores, with sensitivities at 1-2?ppb (ng?mL?1) for biomolecules and 10? to 103 spores per milliliter. We report its application in the detection of acidophilic microorganisms in the Río Tinto Mars analogue and report the absence of substantial negative effects on the immunoassay in the presence of 50?mM perchlorate (20 times higher than that found at the Phoenix landing site). Our SOLID instrument concept is an excellent option with which to detect biomolecules because it avoids the high-temperature treatments that may destroy organic matter in the presence of martian oxidants.     

高温胁迫对岷江百合幼苗耐热指数和理化特性的影响

为了探明岷江百合幼苗在高温胁迫下的生理反应,以离体扩繁的岷江百合幼苗为研究材料,对其进行了不同高温(37 ℃/30 ℃、42 ℃/37 ℃)和不同时间(0、4、8、16、32、48 h)处理,研究了高温胁迫对其耐热指数和有关生理生化指标(叶绿素、丙二醛、游离脯氨酸、可溶性蛋白含量和SOD活性)的影响.结果表明:随着温度的升高和处理时间的延长,植株的耐热指数不断下降,叶绿素含量减少,丙二醛(MDA)含量增加;42 ℃胁迫32 h后,游离脯氨酸、可溶性蛋白含量、SOD活性均达到最大值,分别比对照增加了0.66倍、1.77倍和9.78倍.但胁迫48 h后,指标显著下降,这说明,42 ℃间断胁迫48 h对植株产生了不可逆的的热伤害作用.     

A microbial oasis in the hypersaline Atacama subsurface discovered by a life detector chip: implications for the search for life on Mars

The Atacama Desert has long been considered a good Mars analogue for testing instrumentation for planetary exploration, but very few data (if any) have been reported about the geomicrobiology of its salt-rich subsurface. We performed a Mars analogue drilling campaign next to the Salar Grande (Atacama, Chile) in July 2009, and several cores and powder samples from up to 5?m deep were analyzed in situ with LDChip300 (a Life Detector Chip containing 300 antibodies). Here, we show the discovery of a hypersaline subsurface microbial habitat associated with halite-, nitrate-, and perchlorate-containing salts at 2?m deep. LDChip300 detected bacteria, archaea, and other biological material (DNA, exopolysaccharides, some peptides) from the analysis of less than 0.5?g of ground core sample. The results were supported by oligonucleotide microarray hybridization in the field and finally confirmed by molecular phylogenetic analysis and direct visualization of microbial cells bound to halite crystals in the laboratory. Geochemical analyses revealed a habitat with abundant hygroscopic salts like halite (up to 260?g kg(-1)) and perchlorate (41.13?μg g(-1) maximum), which allow deliquescence events at low relative humidity. Thin liquid water films would permit microbes to proliferate by using detected organic acids like acetate (19.14?μg g(-1)) or formate (76.06?μg g(-1)) as electron donors, and sulfate (15875?μg g(-1)), nitrate (13490?μg g(-1)), or perchlorate as acceptors. Our results correlate with the discovery of similar hygroscopic salts and possible deliquescence processes on Mars, and open new search strategies for subsurface martian biota. The performance demonstrated by our LDChip300 validates this technology for planetary exploration, particularly for the search for life on Mars.     

Assessing antibody microarrays for space missions: effect of long-term storage, gamma radiation, and temperature shifts on printed and fluorescently labeled antibodies

Antibody microarrays are becoming frequently used tools for analytical purposes. A key factor for optimal performance is the stability of the immobilized (capturing) antibodies as well as those that have been fluorescently labeled to achieve the immunological test (tracers). This is especially critical for long-distance transport, field testing, or planetary exploration. A number of different environmental stresses may affect the antibody integrity, such as dryness, sudden temperature shift cycles, or, as in the case of space science, exposure to large quantities of the highly penetrating gamma radiation. Here, we report on the effect of certain stabilizing solutions for long-term storage of printed antibody microarrays under different conditions. We tested the effect of gamma radiation on printed and freeze- or vacuum-dried fluorescent antibodies at working concentrations (tracer antibodies), as well as the effect of multiple cycles of sudden and prolonged temperature shifts on the stability of fluorescently labeled tracer antibody cocktails. Our results show that (i) antibody microarrays are stable at room temperature when printed on stabilizing spotting solutions for at least 6 months, (ii) lyophilized and vacuum-dried fluorescently labeled tracer antibodies are stable for more than 9 months of sudden temperature shift cycles (-20°C to 25°C and 50°C), and (iii) both printed and freeze- or vacuum-dried fluorescent tracer antibodies are stable after several-fold excess of the dose of gamma radiation expected during a mission to Mars. Although different antibodies may exhibit different susceptibilities, we conclude that, in general, antibodies are suitable for use in planetary exploration purposes if they are properly treated and stored with the use of stabilizing substances.     

星载缝隙波导微波天线热控方案研究与验证

微波天线阵面的热变形是影响在轨指向精度的关键因素，如何解决大功率器件的散热问题至关重要。某微波天线热耗峰值近万瓦，在近20 m²的全阵面内，需要保证收发(TR)组件的温升和补偿加热器的功耗不能过高。针对某微波天线特有的工作模式和外热流情况，提出了机械泵驱动流体回路(MPFL)、双面散热和相变热管 3种热控方案，并分析了各自的特点和适用性。为解决天线内部狭小空间的辐射传导耦合问题，开展了单模块热阻实验分析与优化，并得到了在轨测试的验证。     

Protein Microarrays-Based Strategies for Life Detection in Astrobiology

The detection of organic molecules of unambiguous biological origin is fundamental for the confirmation of present or past life. Planetary exploration requires the development of miniaturized apparatus for in situ life detection. Analytical techniques based on mass spectrometry have been traditionally used in space science. Following the Viking landers, gas chromatography-mass spectrometry (GC-MS) for organic detection has gained general acceptance and has been used successfully in the Cassini–Huygens mission to Titan. Microfluidics allows the development of miniaturized capillary electrophoresis devices for the detection of important molecules for life, like amino acids or nucleobases. Recently, a new approach is gaining acceptance in the space science community: the application of the well-known, highly specific, antibody–antigen affinity interaction for the detection and identification of organics and biochemical compounds. Antibodies can specifically bind a plethora of structurally different compounds of a broad range of molecular sizes, from amino acids level to whole cells. Antibody microarray technology allows us to look for the presence of thousands of different compounds in a single assay and in just one square centimeter. Herein, we discuss several important issues—most of which are common with other instruments dealing with life signature detection in the solar system—that must be addressed in order to use antibody microarrays for life detection and planetary exploration. These issues include (1) preservation of biomarkers, (2) the extraction techniques for biomarkers, (3) terrestrial analogues, (4) the antibody stability under space environments, (5) the selection of unequivocal biomarkers for the antibody production, or (6) the instrument design and implementation.     

Classification of modern and old Río Tinto sedimentary deposits through the biomolecular record using a life marker biochip: implications for detecting life on Mars

The particular mineralogy formed in the acidic conditions of the Río Tinto has proven to be a first-order analogue for the acid-sulfate aqueous environments of Mars. Therefore, studies about the formation and preservation of biosignatures in the Río Tinto will provide insights into equivalent processes on Mars. We characterized the biomolecular patterns recorded in samples of modern and old fluvial sediments along a segment of the river by means of an antibody microarray containing more than 200 antibodies (LDCHIP200, for Life Detector Chip) against whole microorganisms, universal biomolecules, or environmental extracts. Samples containing 0.3-0.5?g of solid material were automatically analyzed in situ by the Signs Of LIfe Detector instrument (SOLID2), and the results were corroborated by extensive analysis in the laboratory. Positive antigen-antibody reactions indicated the presence of microbial strains or high-molecular-weight biopolymers that originated from them. The LDCHIP200 results were quantified and subjected to a multivariate analysis for immunoprofiling. We associated similar immunopatterns, and biomolecular markers, to samples with similar sedimentary age. Phyllosilicate-rich samples from modern fluvial sediments gave strong positive reactions with antibodies against bacteria of the genus Acidithiobacillus and against biochemical extracts from Río Tinto sediments and biofilms. These samples contained high amounts of sugars (mostly polysaccharides) with monosaccharides like glucose, rhamnose, fucose, and so on. By contrast, the older deposits, which are a mix of clastic sands and evaporites, showed only a few positives with LDCHIP200, consistent with lower protein and sugar content. We conclude that LDCHIP200 results can establish a correlation between microenvironments, diagenetic stages, and age with the biomarker profile associated with a sample. Our results would help in the search for putative martian biomarkers in acidic deposits with similar diagenetic maturity. Our LDCHIP200 and SOLID-like instruments may be excellent tools for the search for molecular biomarkers on Mars or other planets.     

Cu对铝热法制备的Fe3Al-10％Ni块体纳米晶材料组织和性能影响

通过铝热法制备了Cu含量为2％,4％,6％(质量分数,下同)的Fe3Al-10％ Ni块体纳米晶材料,用光学显微镜(OM),电子探针(EPMA),X-射线衍射仪(XRD)和透射电镜(TEM)表征了材料的微观结构,并测定了材料的维氏硬度和室温压缩性能.结果表明:Cu含量低(2％)时,合金保持无序bcc结构,晶粒尺寸显著下...