How We Are Planning a Facility To Receive Mars Samples
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Alvin L Smith↗
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Evolution of the Lunar Receiving Laboratory to the Astromaterial Sample Curation Facility: Technical Tensions Between Containment and Cleanliness, Between Particulate and Organic Cleanliness
The Lunar Receiving Laboratory (LRL) was planned and constructed in the 1960s to support the Apollo program in the context of landing on the Moon and safely returning humans. The enduring science return from that effort is a result of careful curation of planetary materials. Technical decisions for the first facility included sample handling environment (vacuum vs inert gas), and instruments for making basic sample assessment, but the most difficult decision, and most visible, was stringent biosafety vs ultra-clean sample handling. Biosafety required handling of samples in negative pressure gloveboxes and rooms for containment and use of sterilizing protocols and animal/plant models for hazard assessment. Ultra-clean sample handling worked best in positive pressure nitrogen environment gloveboxes in positive pressure rooms, using cleanable tools of tightly controlled composition. The requirements for these two objectives were so different, that the solution was to design and build a new facility for specific purpose of preserving the scientific integrity of the samples. The resulting Lunar Curatorial Facility was designed and constructed, from 1972-1979, with advice and oversight by a very active committee comprised of lunar sample scientists. The high precision analyses required for planetary science are enabled by stringent contamination control of trace elements in the materials and protocols of construction (e.g., trace element screening for paint and flooring materials) and the equipment used in sample handling and storage. As other astromaterials, especially small particles and atoms, were added to the collections curated, the technical tension between particulate cleanliness and organic cleanliness was addressed in more detail. Techniques for minimizing particulate contamination in sample handling environments use high efficiency air filtering techniques typically requiring organic sealants which offgas. Protocols for reducing adventitious carbon on sample handling surfaces often generate particles. Further work is needed to achieve both minimal particulate and adventitious carbon contamination. This paper will discuss these facility topics and others in the historical context of nearly 50 years' curation experience for lunar rocks and regolith, meteorites, cosmic dust, comet particles, solar wind atoms, and asteroid particles at Johnson Space Center.
Allton, J. H.↗
Mars Sample Return Planning - Strategies for Meeting Planetary Protection Requirements
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Mars Sample Return↗
MSR Sample Receiving Project - Ongoing Activities, Curation, and Facility Planning
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Alvin Smith↗
Tank Waste Characterization: History, Challenges, and Success Stories
The preparation and chemical and radiochemical analysis of Hanford tank waste samples can be performed with standard laboratory equipment and instruments as relatively routine processes that are not particularly challenging. Rather, the main challenges of tank waste characterization are associated with radiological dose and sampling limitations. Accurate, representative and effective sampling techniques are difficult with the waste tanks because they were not designed for routine sampling. There are a finite number of sampling locations for each tank based on riser positioning, depth and the operational functionality of the sampling riser. For example, in one recently emptied SST, there was one riser that was found to have had concrete dumped down it, thereby eliminating that sampling port. Additionally, the waste within the tank; especially true for the saltcake and sludge, is not homogenous. The ability to adequately mix a million-gallon double shell tank (DST) is a concern for data reproducibility. Another real challenge that must be addressed for sampling single shell tanks, is how to dissolve the salt cake waste in a compromised (leaking) SST. These physical constraints mean that uncertainty in the representativeness of samples must be considered when applying analytical results to the bulk contents of the tank. The tank waste is highly radioactive and thus can only be handled initially by facilities that can receive samples into concrete-shielded hot cells with remote operation with an example provided in Figure 1. The shielding protects the worker from the radiological dose while mineral oil windows and remotely operated manipulators enables the samples to be handled. At Hanford, analytical laboratories with these hot cell capabilities are limited to the Pacific Northwest National Laboratory and the main Hanford operations support laboratory, 222-S Laboratory. Because of their highly radioactive nature, samples must be sufficiently diluted to facilitate their analysis outside of a shielded cell. In some cases, this means some accuracy must be compromised to complete the analysis beyond that normally encountered for non-radioactive material.
Waste Characterization, BBI, PHOENIX: Tank Farms: ↗
Curating NASA's Astromaterials Collections: Past, Present, and Future
Planning for the curation of samples from future sample return missions must begin during the initial planning stages of a mission. Waiting until the samples have been returned to Earth, or even when you begin to physically build the spacecraft is too late. A lack of proper planning could lead to irreversible contamination of the samples, which in turn would compromise the scientific integrity of the mission. For example, even though the Apollo missions first returned samples in 1969, planning for the curation facility began in the early 1960s, and construction of the Lunar Receiving Laboratory was completed in 1967. In addition to designing the receiving facility and laboratory that the samples will be characterized and stored in, there are many aspects of contamination that must be addressed during the planning and building of the spacecraft: planetary protection (both outbound and inbound); cataloging, documenting, and preserving the materials used to build spacecraft (also known as coupons); near real-time monitoring of the environment in which the spacecraft is being built using witness plates for critical aspects of contamination (known as contamination control); and long term monitoring and preservation of the environment in which the spacecraft is being built for most aspects of potential contamination through the use of witness plates (known as contamination knowledge). The OSIRIS REx asteroid sample return mission, currently being built, is dealing with all of these aspects of contamination in order to ensure they return the best preserved sample possible. Coupons and witness plates from OSIRIS REx are currently being studied and stored (for future studies) at the Johnson Space Center. Similarly, planning for the clean room facility at Johnson Space Center to house the OSIRIS-REx samples is well advanced, and construction of the facility should begin in early 2017 (despite a nominal 2023 return date for OSIRIS-REx samples). Similar development is being done, in concert with JAXA, for the return of Hayabusa 2 samples (nominally in 2020). We are also actively developing advanced techniques like cold curation and organically clean curation in anticipation of future sample return missions such as comet nucleus sample return and Mars sample return.
Zeigler, Ryan↗
Receiving and Processing Isolators and the Handling of Martian Samples Inside a BSL-4+ Containment Facility: Summary Video Presentation Teaser
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Mars Sample Return; Curation↗
Receiving and Processing Isolators and the Handling of Martian Samples Inside a BSL-4 Containment Facility
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Mars Sample Return↗
Receiving and Processing Isolators and the Handling of Martian Samples Inside a BSL-4 Containment Facility
No abstract provided
McCubbin, Francis↗
Overview of Mars Sample Hazard Analysis (Requirements Workshop Series)
Samples returned from Mars should be contained and treated as though potentially hazardous until proven otherwise. If sample containment cannot be verified en route to Earth, the sample and spacecraft should either be sterilized in space or not returned to Earth. Integrity of sample containment should be maintained through reentry and transfer to a receiving facility. Controlled distribution of unsterilized materials should only occur if analyses determine the sample not to contain a biological hazard. Planetary protection measures adopted for the first sample return should not be relaxed for subsequent missions without thorough scientific review and concurrence by an appropriate independent body.
Rummel, John D.↗
Introduction to Mars Sample Handling Workshop Series: Overview of Mars Sample Hazard Analysis
Samples returned from Mars should be contained and treated as though potentially hazardous until proven otherwise If sample containment cannot be verified en route to Earth, the sample and spacecraft should either be sterilized in space or not returned to Earth. Integrity of sample containment should be maintained through reentry and transfer to a receiving facility. Controlled distribution of unsterilized materials should only occur if analyses determine the sample not to contain a biological hazard. Planetary protection measures adopted for the first sample return should not be relaxed for subsequent missions without thorough scientific review and concurrence by an appropriate independent body
Rummel, John D.↗
Mobile/Modular BSL-4 Containment Facilities Integrated into a Curation Receiving Laboratory for Restricted Earth Return Missions
NASA robotic sample return missions designated Category V Restricted Earth Return by the NASA Planetary Protection (PP) Office require sample containment and biohazard testing upon return to Earth. Since the 1960s, sample containment from an unknown extraterrestrial biohazard have been related to the highest containment standards and protocols known to modern science. Today, this is Biosafety Level (BSL) 4 containment. In the U.S., the Biosafety in Microbiological and Biomedical Laboratories publication authored by the U.S. Department of Health and Human Services (HHS): Public Health Service, Centers for Disease Control and Prevention, and the National Institutes of Health houses the primary recommendations, standards, and design requirements for all BSL labs. Past mission concept studies for constructing a NASA Curation Receiving Laboratory with an integrated BSL-4 quarantine and biohazard testing facility have been estimated in the hundreds of millions of dollars (USD). As an alternative option, we have conducted a trade study for constructing a mobile and/or modular sample containment laboratory that would meet all BSL-4 and planetary protection standards and protocols at a fraction of the cost. Mobile and modular BSL-2 and 3 facilities have been successfully constructed and deployed world-wide for government testing of pathogens and pharmaceutical production. Our study showed that a modular BSL-4 construction could result in ~ 90% cost reduction when compared to traditional BSL-4 construction methods without compromising the preservation of the samples or Earth. For the design/construction requirements of a mobile/modular BSL-4 containment, we used the established HHS document standards and protocols for manipulation of agents in Class III Biosafety Cabinets (BSC; i.e., negative pressure gloveboxes) that are currently followed in operational BSL-4 facilities in the U.S.
Calaway, Michael J.↗
Developing the New Hayabusa Curation Facility at Johnson Space Center
On 25 November 2005 the Japan Aerospace Exploration Agency (JAXA) Hayabusa spacecraft made contact with the asteroid 25143 Itokawa and collected a small amount of regolith dust from Muses Sea region of smooth terrain [1]. Even though optimal sample collection did not occur, the spacecraft returned to Earth with more than 10,000 grains ranging in the size from 30-180 microns [2]. These grains represent the only collection of pristine material returned from an asteroid by a spacecraft. As part of the joint agreement between JAXA and NASA for the mission, 10% of the Hayabusa grains will be transferred to NASA for parallel curation and allocation, the first 15 of which arrived in December 2011. In order to properly receive and process these samples, a new curation facility was developed at Johnson Space Center (JSC).
De Gregorio, B. T.↗
The Hayabusa Curation Facility at Johnson Space Center
The Japan Aerospace Exploration Agency (JAXA) Hayabusa spacecraft made contact with the asteroid 25143 Itokawa and collected regolith dust from Muses Sea region of smooth terrain [1]. The spacecraft returned to Earth with more than 10,000 grains ranging in size from just over 300 μm to less than 10 μm [2, 3]. These grains represent the only collection of material returned from an asteroid by a spacecraft. As part of the joint agreement between JAXA and NASA for the mission, 10% of the Hayabusa grains are being transferred to NASA for parallel curation and allocation. In order to properly receive process and curate these samples, a new curation facility was established at Johnson Space Center (JSC). Since the Hayabusa samples within the JAXA curation facility have been stored free from exposure to terrestrial atmosphere and contamination [4], one of the goals of the new NASA curation facility was to continue this treatment. An existing lab space at JSC was transformed into a 120 sq.ft. ISO class 4 (equivalent to the original class 10 standard) clean room. Hayabusa samples are stored, observed, processed, and packaged for allocation inside a stainless steel glove box under dry N2. Construction of the clean laboratory was completed in 2012. Currently, 25 Itokawa particles are lodged in NASA's Hayabusa Lab. Special care has been taken during lab construction to remove or contain materials that may contribute contaminant particles in the same size range as the Hayabusa grains. Several witness plates of various materials are installed around the clean lab and within the glove box to permit characterization of local contaminants at regular intervals by SEM and mass spectrometry, and particle counts of the lab environment are frequently acquired. Of particular interest is anodized aluminum, which contains copious sub-mm grains of a multitude of different materials embedded in its upper surface. Unfortunately the use of anodized aluminum was necessary in the construction of the clean room frame to strengthen it and eliminate corrosion and wear over time. All anodized aluminum interior to the lab was thus covered or replaced by minimally contaminating materials.
Zolensky, M.↗
Analytical Results of the Tank 51H Sludge Batch 11 As-Received Qualification Sample
Savannah River National Laboratory (SRNL) has been requested by Savannah River Mission Completion (SRMC) to qualify the next sludge batch, Sludge Batch 11 (SB11), for processing at the Defense Waste Processing Facility (DWPF). This report documents the first steps of the qualification process, characterization of the Tank 51H as-received qualification material. SRMC sent SRNL two 200 mL samples received from Tank 51H on the 12th of August 2024. The combined sample, HTF-51-24-85/86, was analyzed for the following: supernate and slurry density, weight percent solids, settling, chemical composition, and radionuclides.
12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
Microbial Monitoring of New Cleanrooms Used to Curate Astrobiologically Relevant Asteroid Samples from Bennu and Ryugu
Introduction: NASA has constructed two new cleanrooms to house materials from the OSRIS-REx and Hayabusa2 missions to the asteroids Ryugu (162173) and Bennu (101955), respectively. In accordance with standard astromaterials curation practices, these cleanrooms will be monitored for particulate contamination and maintained to ISO 5 equivalent standards1. Since the samples in these collections are expected to contain prebiotic organic compounds that may help explain the origin of life on Earth, these labs will also be monitored for organic and biological contamination2. Samples from Ryugu arrived on Earth in December, 2020. After basic characterization in Japan, NASA received a subset of these samples at the astromaterials curation facility in Houston in December of 2021. OSIRIS-REx is expected to return samples in September, 2023. Here we present preliminary microbial monitoring results from monthly monitoring of these new labs and the connected microtomy and staging areas that support them, as they are being commissioned. We also compare these results to baseline values for other astromaterials curation labs. We will also briefly describe additional cleaning efforts employed to reduce the bioburden in these new cleanrooms. Methods: Microbial samples were collected from surfaces using a dry macrofoam swab (Puritan Brand 2518051PFRNDFD). Swabs were also opened in the lab but not touched to any surfaces to function as negative controls. Samples and controls were processed inside a class II biosafety cabinet to avoid inadvertent cross contamination. The swabs were suspended in 15 ml of PBS (Phosphate Buffered Saline) and vortexed for 20 seconds to remove cells from the swab surface. The PBS was used to inoculate Petri dishes filled with TSA (Tryptic Soy Agar), Blood Agar, or Reasoners 2 agar to check for microbial growth. Each plate was inoculated with 0.1 ml of PBS. The TSA and blood agar plates were incubated at 35˚C and the Reasoners 2 agar plates were incubated at 25˚C for seven days. Petri dishes filled with Potato dextrose agar, Saboraud dextrose agar, or Saboraud dextrose agar with 0.1 mg/ml of chloramphenicol, an antibiotic, were used to check for fungal growth. These plates were inoculated with 0.3 ml of PBS and incubated at 30˚C. The remaining PBS was frozen at -80 ˚C for DNA sequencing. After incubation, isolates were counted and reisolated for identification. Isolates were identified using the VITEK23 system or by sequencing a portion of the 16S rRNA gene for bacteria or the ribosomal internal transcribed spacer (ITS) for fungi. Sequencing was performed with an ABI 3500 Sanger sequencer. Results: During our initial sampling, six of the seven sites sampled (86%) displayed bacterial or fungal growth. Samples collected from the staging areas and microtomy labs are not included in this calculation since those areas are maintained at a lower ISO 7 equivalent cleanliness standard. A month later, only three of the seven sites (43%) displayed bacterial growth. No fungal growth was detected in the second sampling. Since new equipment had been introduced to the Hayabusa2 lab since the first round of sampling, an additional three sampling sites were included in the second round of sampling. None of these sites displayed microbial growth. These sites will be included in all future sampling efforts. Bacterial isolates have been identified from the following genera at multiple time points: Micrococcus, Staphylococcus, and Bacillus. Isolates from the genera: Microbacterium, Nocardioides, Methylocystis, and Microvirga were identified in the initial sampling, but were not present at later time points. Identification of fungal isolates is in progress. Results are summarized in Table 1. Discussion: The recovery rate or percentage of positive samples4 was initially 86%, which is higher than the median recovery rate for comparable ISO 5 equivalent curation labs like Stardust (33%), Hayabusa (33%), and Cosmic Dust (50%). However, after a month of operation, the recovery rate for these same sites decreased to 43%, which is similar to what we observe in comparable curation cleanrooms with no microbial control requirements. Adding in the new sampling sites further decreases the recovery rate to 30%. With the reduction in recovery rate, we also observed a decrease in microbial diversity. At the first time point, we observed at least 10 different bacterial species and at least two different fungi. This is a higher diversity than the median values for comparable ISO 5 equivalent labs (2-4 isolates per sampling event). After the second sampling, we observed at least 4 bacterial species and no fungi, which is more consistent with comparable labs. We expect the recovery rate and diversity in both labs to continue to decrease as routine operation continues. We will use ultrapure hydrogen peroxide to disinfect equipment and work areas prior to opening any sample containers. Most of the bacterial and fungal isolates were detected on samples from the cleanroom floors. This is consistent with baseline results from other curation labs. Organisms from the genera Bacillus, Staphylococcus, and Micrococcus that were repeatedly detected are common in cleanrooms and on human skin5,6. These organisms are generally thought to be introduced when people enter the cleanroom. Microbacterium, Nocardioides, and Microvirga have also previously been identified in astromaterials cleanrooms, but not as frequently as Bacillus, Staphylococcus, and Micrococcus. Methylocystis is a novel genus in the astromaterials cleanrooms, but it was identified with low accuracy (93% match in the sequenced region of the 16S rRNA gene) and further work is needed to confirm this identification. Microbacterium is a diverse genus with isolates identified from terrestrial and aquatic sediments. Some species of Microbacterium are capable of degrading complex organic compounds found in crude oil. The presence of these bacteria in the OSIRIS REx and Hayabusa2 cleanrooms should be closely monitored. Methylocystis is a genus of methanotrophic bacteria capable of oxidizing methane. If this identification proves to be correct and it is detected again, it should be closely monitored as well. Under nominal operating conditions, samples should not ever encounter the cleanroom floor or other high traffic areas. If we observe an increase in the bioburden in sensitive work areas that appears to be influenced by organism transfer from high traffic areas like the floors, we can employ additional hydrogen peroxide treatments to disinfect high traffic areas. Routine microbial monitoring of these labs will ensure that NASA’s astromaterials collections remain pristine and useful for scientific study. Table 1. Sampling Locations and Colony Counts Bacterial CFUa Fungal CFU Bacterial CFU Fungal CFU Lab - Location 11/2/2021 11/2/2021 12/13/2021 12/13/2021 H2b-Floor 4 8 1 0 H2-staging pass through 3 0 0 0 H2-microtomy pass through TNTCc 0 0 0 H2 Microscope 1 NA NA 0 0 H2 Microscope 2 NA NA 0 0 H2-Table NA NA 0 0 OREXd- microtomy pass through 0 0 6 0 OREX – Anteroom pass through 0 0 0 0 OREX – Floor 1 2 0 0 OREX Witness Foil Table 3 0 1 0 Staging-Floor 16 0 15 0 Microtomy-Floor 3 0 2 0 a: CFU = Colony Forming Unit b: H2 = Hayabusa2 Lab c: TNTC = too numerous to count d: OREX = OSIRIS-REx Lab References: 1. ISO 14644-1:2015 - Cleanrooms and associated controlled environments -- Part 1: Classification of air cleanliness by particle concentration. 37 (2015). 2. McCubbin, F. M. et al. Space Sci Rev 215, (2019). 3. Pincus, D. H. Encyclopedia of Rapid Microbiological Methods (2005). 4. The United States Pharmacopeial Convention. USP General Chapter <1116> 17, 784–794 (2013). 5. Sheraba, N. S., Yassin, A. S. & Amin, M. BMC Research Notes 3, 278 (2010). 6. Utescher, C. L. de A., Franzolin, M. R., Trabulsi, L. R. & Gambale, V. Brazilian Journal of Microbiology 38, 710–716 (2007).
A B Regberg↗
Initial analysis of “stone” size Ryugu samples: current status
As a part of the initial analysis of the Ryugu samples, we perform a variety of analyses of millimeter-sized "stones". Our goals are to elucidate the entire formation process of C-type asteroid Ryugu from the viewpoint of petrology and mineralogy ande obtain necessary information by sample analysis, and then simulate the formation of Ryugu based on the evidencef obtained rom sample analysis. Eighteen stones (8 from Room A and 10 from Room C) were received from the ISAS curation facility on June 1, 2021, and brought into a fully nitrogen-displaced glove box at Tohoku University. At the same time, we also received the powder samples from Room A and Room C. To completely block the atmosphere from leaking into the container, all samples were put in the sample transport containers prepared by ISAS, transferred from the main chamber to the glovebox at ISAS, and then all containers were completely sealed in plastic bags with moisture and oxygen absorbers. No moisture or oxygen was detected when the bags were opened in the glove box at Tohoku University, so it was confirmed that there was no exposure to the atmosphere during transport. To date, a number of analyses have been carried out successfully and almost on schedule. The analysis started with the measurement of reflectance spectra, which are sensitive to atmospheric oxidation, hydroxylation, and adsorbed water. The visible, near-infrared, and mid-infrared reflectance spectra were measured while the samples were kept airtight. The spectra of powder samples and stone samples (as aggregates and as single stone) were successfully obtained. A major feature of the stone team's analysis is the use of synchrotron radiation facilities around the world. Since this analysis is non-destructive, stone samples whose reflectance spectra were measured were sent to KEK, SPring-8, ESRF (France), SOLEIL (France), DESY (Germany), and APS (USA). Using these synchrotron radiation facilities, high spatial resolution and sensitivity XRD, STXM, XANES, CT [1], IR-CT, FT-IR [2, 3], XRF, and Mössbauer analyses were performed. Most of the analyses were carried out under air-tight conditions on the stone samples and the particulates separated from the stone samples. These analyses allowed us to determine the three-dimensional distribution of minerals and elements, redox state, density and porosity of the stone samples. Furthermore, as a characteristic analysis of the stone team, light elemental analysis using negative Muon was performed at the MLF facility of J-PARC with an exceptionally long allocation of machine time. This is the only non-destructive method to measure the concentration of light elements in the whole (not the surface) of stone samples. Because the characteristic X-rays produced by muon irradiation are much higher in energy than the fluorescent X-rays produced by X-ray irradiation, there is little effect of self-absorption by the sample, and therefore, the concentration of light elements such as carbon, oxygen, and Na in the entire "stone" sample can be determined. Some stone samples are currently being measured for heat and strength physical properties in order to understand the physical properties of asteroid Ryugu. The data obtained from these measurements are useful for interpreting the remote-sensing data data taken from the surface layer of the asteroid Ryugu [4-7]. It is also important for understanding the behavior of the Ryugu material during impact events. The surfaces of many stone samples were observed by electron microscopy and other techniques, especially on natural “flat” surfaces formed on 5 stomes. As a result, characteristric mineral aggregates formed by the reactions with water and characteiristic impact features were observed on some of the samples. Based on the observations of surfaces and the synchrotron measurements of the whole stones, important objects such as characteristic structures and specific crystal aggregates for understanding the formation history of the asteroid were identified, and these parts were separated from the stone samples using an Xe beam (pFIB) and analyzed by various methods including transmission electron microscopy and synchrotron radiation analysis. Many stone samples, from which important objects have been separated, are embedded in epoxy resin and cut to produce many polished sections. Electron microscopy and spectroscopic measurements of the polished surfaces are being carried out to reveal the detailed mineralogical properties and elemental distribution inside the stone samples. In the fall, machine time for synchrotron radiation will begin, and we plan to analyze single crystals and characteristic objects separated from the stone samples.
Tomoki Nakamura↗