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Project Progress

Co-Investigator Steele and colleagues have continued to develop the Modular Assays for Solar System Exploration (MASSE) concept, which uses microfluidic technology to incubate a DNA or protein microarray. Specifically this year they developed models to aid in the filling of a microarray chamber from microfluidic channels; continued the testing of antibodies in simulated spaceflight environments (in collaboration with University of Leicester); designed, built, and tested a miniaturized imaging system capable of imaging a small microarray using planar waveguide technology; designed, fabricated, and tested a microfluidic microarray chip [in collaboration with the Lab-On-a-Chip Applications and Development (LOCAD) group at Marshall Space Flight Center]; designed and started to fabricate a portable unit to interface with the designed microfluidic microarray device to enable interrogation of a 400-spot protein microarray (in collaboration with LOCAD); designed, built, and tested a portable device to interrogate a sample using a limited number of microarray spots [in collaboration with Charles River Laboratories (CRL)]; tested a number of antibody immobilization techniques, surface functionalization, and labeling protocols for optimal performance of a microarray in a microfluidic system (in collaboration with CRL); designed and are currently testing a DNA microarray to interrogate a sample for the presence of DNA from specific organisms of interest to astrobiology and human spaceflight; and continued the testing of a microbial detection system, named Portable Test System (PTS), which will fly to the International Space Station (ISS) on STS 116A. This effort (in collaboration with CRL and LOCAD) will fly the first miniaturized instrument for the detection of microbial load on the ISS.

The MASSE team has collaborated with the SMILE (Specific Molecular Identification of Life Experiment) team, led by Mark Sims at the University of Leicester, which has successfully proposed to build the life marker chip (LMC) portion of the European Space Agency (ESA) ExoMars mission. The MASSE team (CIW, LOCAD, and CRL) is currently collaborating with SMILE on proof-of-concept studies for ESA and will apply for funding to aid in the development of a flight unit.

To highlight three of the above-mentioned areas of work:

(1) High-density oligonucleotide microarrays allow researchers to interrogate simultaneously large quantities of genetic information in a single experiment. DNA microarrays have been used with success to conduct whole-genome analysis of single-nucleotide polymorphisms (SNPs), generate maps of transcriptional activity across the entire genomes, and resequence non-contiguous regions of genomic DNA. Previous studies have demonstrated that genomic DNA from pathogens can be resequenced using high-density oligonucleotide microarrays. Through the analysis of DNA sequence variation, multiple pathogens can be identified in a single assay with high sensitivity and specificity. We have designed a high-density oligonucleotide microarray for the identification of microorganisms from diverse environments that are applicable to space exploration. Specifically, Steele and his colleagues have designed a custom Affymetrix GeneChip® resequencing array to interrogate the 16S or 18S rRNA and rpoB gene sequences in approximately 120 organisms. The targeted array design includes gene sequences for pathogens, extremophiles, and common microorganisms from the three domains of life — bacteria, archaea, and eukaryota. The selected organisms have potential relevance to specific areas of space exploration, including bio-regenerative environmental control and life-support systems (ECLSS), crew health, in situ resource utilization (ISRU), and techniques for the purposes of life detection (LD) and planetary protection (PP). Microorganisms interfere with or are crucial to the above-mentioned goals and systems, and consequently the development of effective microbial monitoring technologies is critical for mission safety and success.

(2) A field-deployable unit that can interrogate a sample using a 4-antibody microarray (Figure 1) was tested on the Arctic Mars Analogue Svalbard Expedition (AMASE) this year. The antibodies chosen monitored for the presence of contaminant bacteria associated with human handling and lipopolysaccharides and were used as a control for contamination during sampling of ice cores, specifically an ice core associated with carbonate caves within the Sverrefjell volcano, which are the sources for a population of ALH84001-type carbonate globules found in basaltic rocks.

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(3) Building on research performed at Meteor Crater, Arizona, in 2005 (Figure 2), LOCAD Project Scientist Jake Maule will return to nearby Cinder Lake in September 2006 to carry out further tests with the LOCAD PTS. The Cinder Lake site (actually not a lake) is somewhat similar to the lunar surface in having dips and hollows with dusty soil. In response to the recent NASA RFI regarding what to do on the Moon in 2018 and beyond, Steele and colleagues proposed that an instrument such as LOCAD PTS be used to monitor how expeditions bio-contaminate lunar and planetary surfaces. In Desert Research and Technology Study (D-RATS) 2006, Maule will integrate LOCAD PTS tests into the Extra-Vehicular Activity (EVA) procedures of two crewmembers who will be simulating a search for in situ resources, e.g., sub-surface water. The 2005 results demonstrated that a crewmember in a fully pressurized suit could operate the LOCAD PTS swabbing unit and perform certain procedures even when dexterity was limited by the suit glove. The 2006 tests will expand upon these initial results, integrating LOCAD PTS further into the operational procedures of a typical surface EVA.

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The AMASE fieldwork in August 2005 also tested several astrobiotechnology-relevant instruments to measure lipopolysaccharide and b-glucan for the presence of gram-negative bacteria and molds, adenosine triphosphate measurements, and functional /16S DNA amplification equipment for rapid detection and characterization of microbial abundances

Planning for AMASE 2006 has been ongoing since January and has received Astrobiology Science and Technology for Exploring Planets (ASTEP) funding (for which Steele is the PI). Steele and colleagues will return to Svalbard to test instruments involved in the Mars Science Laboratory (MSL) mission in 2009; specifically the CheMin (for CHEmistry and MINeralogy) (David Blake, NASA Ames Research Center, PI) and Sample Analysis of Mars (SAM) (Paul Mahaffy, Goddard Space Flight Center, PI) instruments, which make up the analytical suite of this mission. Furthermore, Steele and colleagues will deploy Raman and deep ultraviolet fluorescence instrumentation provided by Pamela Conrad (JPL) and a portable version of the Life Marker Chip instrument being developed for ESA ExoMars. Alongside these instruments will be sample processing and handling equipment, including a drill corer and a rock crusher concept designed for MSL. This equipment will be tested for cleanliness and cross contamination with several biotechnology instruments that have been deployed in the field for the last 3 years. Steele and colleagues will be deploying these instruments alongside a “Cliffbot” rover that has the ability to access steep cliffs and sample from areas that the current Mars Exploration Rover design could not reach. The teams will interact and be trained in how to conduct science and sampling protocols with a rover crew and make science decisions on which investigations to undertake on a particular sample in as complete a mission scenario as possible. Steele and his team will also undertake the testing of the “Cliffbot” alongside an astronaut wearing a Mark 3 space suit in an effort to simulate manned mission goals for detecting life using a rover to reach difficult samples.