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At least 127 records · Page 7

WetLab-2: Tools for Conducting On-Orbit Quantitative Real-Time Gene Expression Analysis on ISS

The objective of NASA Ames Research Centers WetLab-2 Project is to place on the ISS a research platform capable of conducting gene expression analysis via quantitative real-time PCR (qRT-PCR) of biological specimens sampled or cultured on orbit. The project has selected a Commercial-Off-The-Shelf (COTS) qRT-PCR system, the Cepheid SmartCycler and will fly it in its COTS configuration. The SmartCycler has a number of advantages including modular design (16 independent PCR modules), low power consumption, rapid ramp times and the ability to detect up to four separate fluorescent channels at one time enabling multiplex assays that can be used for normalization and to study multiple genes of interest in each module. The team is currently working with Cepheid to enable the downlink of data from the ISS to the ground and provide uplink capabilities for programming, commanding, monitoring, and instrument maintenance. The project has adapted commercial technology to design a module that can lyse cells and extract RNA of sufficient quality and quantity for use in qRT-PCR reactions while using a housekeeping gene to normalize RNA concentration and integrity. The WetLab-2 system is capable of processing multiple sample types ranging from microbial cultures to animal tissues dissected on-orbit. The ability to conduct qRT-PCR on-orbit eliminates the confounding effects on gene expression of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of fixed samples. The system can be used to validate terrestrial analyses of samples returned from ISS by providing on-orbit gene expression benchmarking prior to sample return. The ability to get on orbit data will provide investigators with the opportunity to adjust experiment parameters for subsequent trials based on the real-time data analysis without need for sample return and re-flight. Researchers will also be able to sample multigenerational changes in organisms. Finally, the system can be used for analysis of air, surface, water, and clinical samples to monitor environmental contaminants and crew health. The verification flight of the instrument is scheduled to launch on SpaceX-7 in June 2015.

quantitative PCR↗

Calibration of TOMS Radiances From Ground Observations

Verification of a stratospheric ozone recovery remains a high priority for environmental research and policy definition. Models predict an ozone recovery at a much lower rate than the measured depletion rate observed to date. Therefore improved precision of the satellite and ground ozone observing systems are required over the long term to verify its recovery. We show that validation of radiances from the ground can be a very effective means for correcting long term drifts of backscatter type satellite measurements and can be used to cross calibrate all BUV instruments in orbit (TOMS, SBUV/2, GOME, SCIAMACHY, OMI, GOME-2, OMPS). This method bypasses the retrieval algorithms used to derive ozone products from both satellite and ground based measurements that are normally used to validate the satellite data. Radiance comparisons employ forward models, but they are inherently more accurate than the retrieval This method employs very accurate comparisons between ground based zenith sicy radiances and satellite nadir radiances and employs two well established capabilities at the Goddard Space Flight Center, 1) the SSBUV calibration facilities and 2) the radiative transfer codes used for the TOMS and SBUV/2 algorithms and their subsequent refinements. The zenith sky observations are made by the SSBUV where its calibration is maintained to a high degree of accuracy and precision. Radiative transfer calculations show that ground based zenith sky and satellite nadir backscatter ultraviolet comparisons can be made very accurately under certain viewing conditions. Initial ground observations taken from Goddard Space Flight Center compared with radiative transfer calculations has indicated the feasibility of this method. The effect of aerosols and varying ozone amounts are considered in the model simulations and the theoretical comparisons. The radiative transfer simulations show that the ground and satellite radiance comparisons can be made with an uncertainty of less than l\% without the knowledge of the amount ozone viewed by either instrument on ground or in space. algorithms.

Bojkov, B. R.↗

Probable Rotation States of Rocket Bodies in Low Earth Orbit

In order for Active Debris Removal to be accomplished, it is critically important to understand the probable rotation states of orbiting, spent rocket bodies. As compared to the question of characterizing small unresolved debris, in this problem there are several advantages: (1) objects are of known size, mass, shape and color, (2) they have typically been in orbit for a known period of time, (3) they are large enough that resolved images may be obtainable for verification of predicted orientation, and (4) the dynamical problem is simplified to first order by largely cylindrical symmetry. It is also nearly certain for realistic rocket bodies that internal friction is appreciable in the case where residual liquid or, to a lesser degree, unconsolidated solid fuels exist. Equations of motion have been developed for this problem in which internal friction as well as torques due to solar radiation, magnetic induction, and gravitational gradient are included. In the case of pure cylindrical symmetry, the results are compared to analytical predictions patterned after the standard approach for analysis of symmetrical tops. This is possible because solar radiation and gravitational torques may be treated as conservative. Agreement between results of both methods ensures their mutual validity. For monotone symmetric cylinders, solar radiation torque vanishes if the center of mass resides at the geometric center of the object. Results indicate that in the absence of solar radiation effects, rotation states tend toward an equilibrium configuration in which rotation is about the axis of maximum inertia, with the axis of minimum inertia directed toward the center of the earth. Solar radiation torque introduces a modification to this orientation. The equilibrium state is asymptotically approached within a characteristic timescale given by a simple ratio of relevant characterizing parameters for the body in question. Light curves are simulated for the expected asymptotic final rotation states of model objects, and these are compared to data derived from physical models of the same objects, tested in the Optical Measurements Center at JSC. Comparison to relevant light curves from actual orbiting rocket bodies are also performed, and diagnostic features of such curves are examined.

Ojakangas, Gregory W.↗

Verification and Validation: High Charge and Energy (HZE) Transport Codes and Future Development

In the present paper, we give the formalism for further developing a fully three-dimensional HZETRN code using marching procedures but also development of a new Green's function code is discussed. The final Green's function code is capable of not only validation in the space environment but also in ground based laboratories with directed beams of ions of specific energy and characterized with detailed diagnostic particle spectrometer devices. Special emphasis is given to verification of the computational procedures and validation of the resultant computational model using laboratory and spaceflight measurements. Due to historical requirements, two parallel development paths for computational model implementation using marching procedures and Green s function techniques are followed. A new version of the HZETRN code capable of simulating HZE ions with either laboratory or space boundary conditions is under development. Validation of computational models at this time is particularly important for President Bush s Initiative to develop infrastructure for human exploration with first target demonstration of the Crew Exploration Vehicle (CEV) in low Earth orbit in 2008.

Wilson, John W.↗

Mars Small-Spacecraft Human Exploration Resource Prospector with Aero-braking (SHERPA): Demonstrating an End-to-End Mission to Phobos Distant Retrograde Orbit

This paper summarizes an end-to-end mission design concept exploring the feasibility of using small satellites together with aero-capture technology to achieve Mars orbit insertion, and subsequent injection into a Phobos-stabilized (or distant retrograde) orbit. The science and mission objectives are to carry out a survey of the mineralogy and morphology of Phobos, to answer basic questions concerning its origin and formation, to test the cohesiveness of Phobos regolith, and to search for potential landing sites for future human or robotic spacecraft. The Mars Small-Spacecraft Human Exploration Resource Prospector with Aero-braking (SHERPA) spacecraft is based on a combination flight-tested prototype vehicle and instruments, and first principle sizing of consumables. The resulting system is fitted with an inflatable aerodynamic decelerator to effect aero-capture into a Mars elliptical orbit, on its way to achieving Phobos orbit. A computational fluid dynamics tool is used to analyze the flow-field and identify potential hot spots during aerodynamic flight. This work advocates for the use of small satellites to test out technologies and operational concepts used in sustained human exploration of Mars, and to carry out scientific exploration of the Mars system. Consistent with a systems engineering approach, this work combines elements of the NASA Human Exploration and Operations Mission Directorate, the Space Technology Mission Directorate, and the Science Mission Directorate, and proposes a scenario for science acquisition, technology verification, trajectory validation, and in-situ resource exploration. We believe these type of missions are essential forerunners to human crewed missions to Mars.

Mission Design↗

Validation of SCIAMACHY and TOMS UV Radiances Using Ground and Space Observations

Verification of a stratospheric ozone recovery remains a high priority for environmental research and policy definition. Models predict an ozone recovery at a much lower rate than the measured depletion rate observed to date. Therefore improved precision of the satellite and ground ozone observing systems are required over the long term to verify its recovery. We show that validation of satellite radiances from space and from the ground can be a very effective means for correcting long term drifts of backscatter type satellite measurements and can be used to cross calibrate all B W instruments in orbit (TOMS, SBW/2, GOME, SCIAMACHY, OM, GOME-2, OMPS). This method bypasses the retrieval algorithms used for both satellite and ground based measurements that are normally used to validate and correct the satellite data. Radiance comparisons employ forward models and are inherently more accurate than inverse (retrieval) algorithms. This approach however requires well calibrated instruments and an accurate radiative transfer model that accounts for aerosols. TOMS and SCIAMACHY calibrations are checked to demonstrate this method and to demonstrate applicability for long term trends.

Hilsenrath, E.↗

Thermal Design of the Instrument for the Transiting Exoplanet Survey Satellite

TESS observatory is a two year NASA Explorer mission which will use a set of four cameras to discover exoplanets. It will be placed in a high-earth orbit with a period of 13.7 days and will be unaffected by temperature disturbances caused by environmental heating from the Earth. The cameras use their stray-light baffles to passively cool the cameras and in turn the CCD's in order to maintain operational temperatures. The design has been well thought out and analyzed to maximize temperature stability. The analysis shows that the design keeps the cameras and their components within their temperature ranges which will help make it a successful mission. It will also meet its survival requirement of sustaining exposure to a five hour eclipse. Official validation and verification planning is underway and will be performed as the system is built up. It is slated for launch in 2017.

Thermal Engineering↗

Credibility Assessment of Deterministic Computational Models and Simulations for Space Biomedical Research and Operations

Human missions beyond low earth orbit to destinations, such as to Mars and asteroids will expose astronauts to novel operational conditions that may pose health risks that are currently not well understood and perhaps unanticipated. In addition, there are limited clinical and research data to inform development and implementation of health risk countermeasures for these missions. Consequently, NASA's Digital Astronaut Project (DAP) is working to develop and implement computational models and simulations (M&S) to help predict and assess spaceflight health and performance risks, and enhance countermeasure development. In order to effectively accomplish these goals, the DAP evaluates its models and simulations via a rigorous verification, validation and credibility assessment process to ensure that the computational tools are sufficiently reliable to both inform research intended to mitigate potential risk as well as guide countermeasure development. In doing so, DAP works closely with end-users, such as space life science researchers, to establish appropriate M&S credibility thresholds. We will present and demonstrate the process the DAP uses to vet computational M&S for space biomedical analysis using real M&S examples. We will also provide recommendations on how the larger space biomedical community can employ these concepts to enhance the credibility of their M&S codes.

Mulugeta, Lealem↗

Experimental Results from the Thermal Energy Storage-2 (TES-2) Flight Experiment

Thermal Energy Storage-2 (TES-2) is a flight experiment that flew on the Space Shuttle Endeavour (STS-72), in January 1996. TES-2 originally flew with TES-1 as part of the OAST-2 Hitchhiker payload on the Space Shuttle Columbia (STS-62) in early 1994. The two experiments, TES-1 and TES-2 were identical except for the fluoride salts to be characterized. TES-1 provided data on lithium fluoride (LiF), TES-2 provided data on a fluoride eutectic (LiF/CaF2). Each experiment was a complex autonomous payload in a Get-Away-Special payload canister. TES-1 operated flawlessly for 22 hr. Results were reported in a paper entitled, Effect of Microgravity on Materials Undergoing Melting and Freezing-The TES Experiment, by David Namkoong et al. A software failure in TES-2 caused its shutdown after 4 sec of operation. TES-1 and 2 were the first experiments in a four experiment suite designed to provide data for understanding the long duration microgravity behavior of thermal energy storage salts that undergo repeated melting and freezing. Such data have never been obtained before and have direct application for the development of space-based solar dynamic (SD) power systems. These power systems will store energy in a thermal energy salt such as lithium fluoride or a eutectic of lithium fluoride/calcium difluoride. The stored energy is extracted during the shade portion of the orbit. This enables the solar dynamic power system to provide constant electrical power over the entire orbit. Analytical computer codes were developed for predicting performance of a space-based solar dynamic power system. Experimental verification of the analytical predictions were needed prior to using the analytical results for future space power design applications. The four TES flight experiments were to be used to obtain the needed experimental data. This paper will address the flight results from the first and second experiments, TES-1 and 2, in comparison to the predicted results from the Thermal Energy Storage Simulation (TESSIM) analytical computer code. An analysis of the TES-2 data was conducted by Cleveland State University Professor, Mounir Ibrahim. TESSIM validation was based on two types of results; temperature history of various points on the containment vessel and TES material distribution within the vessel upon return from flight. The TESSIM prediction showed close comparison with the flight data. Distribution of the TES material within the vessel was obtained by a tomography imaging process. The frozen TES material was concentrated toward the colder end of the canister. The TESSIM prediction indicated a similar pattern. With agreement between TESSIM and the flight data, a computerized representation was produced to show the movement and behavior of the void during the entire melting and freezing cycles.

Tolbert, Carol↗

The Solid Rocket Motor Slag Population: Results of a Radar-Based Regressive Statistical Evaluation

Solid rocket motor (SRM) slag has been identified as a potential source of man-made orbital debris. The possibility that SRMs (in addition to generating dust particles in the sub-millimeter range) may generate particles up to centimeters in size has caused concern regarding their contribution to the debris environment. Returned surfaces from space do not have sufficient area or exposure time to provide a clear picture of the SRM millimeter and centimeter debris population. Currently, radar observation is probably the only way to collect data showing the debris contribution from SRMs. Such observation is used to sample the debris environment, but it is difficult to obtain accurate orbital elements for the detected debris objects. NASA has developed several models to describe the different orbital debris populations, based on assumed debris production mechanisms to create clouds of debris objects that can be propagated in time. The NASA model, LEGEND (LEO-to-GEO Environment Debris), functions as a time-tested debris model for most debris sources. However, the current LEGEND model does not include contributions from the SRM population. An SRM model has recently been developed by NASA, based on purely theoretical details of SRM production and known SRM launches, but verification with hard data is needed. Because the detections of individual SRM objects cannot be deterministically separated from the total debris observed by radar, the validation of the SRM model can only be done by combining it with the LEGEND breakup model and comparing it with data. By applying observational constraints, the degree of SRM slag contribution to the environment may be estimated. This serves as an observationally sound method from which to calibrate a purely theoretical model into something more realistic. For this study, we use the populations observed by the Haystack radar from 1996 to present. For the SRM debris, we use a historical database of SRM launches, propellant masses, and estimated locations and times of tailoff to produce and propagate the SRM debris clouds. Comparisons with radar data from the ensuing years were made, and the SRM model was altered with respect to size and mass production of slag particles to reflect the populations estimated from the data. The result is a model SRM population that fits within the bounds of the observed environment and estimates of the production and contribution of SRM debris to the environment.

Horstman, Matthew F.↗

SEDS1 mission software verification using a signal simulator

The first flight of the Small Expendable Deployer System (SEDS1) is schedule to fly as the secondary payload of a Delta 2 in March, 1993. The objective of the SEDS1 mission is to collect data to validate the concept of tethered satellite systems and to verify computer simulations used to predict their behavior. SEDS1 will deploy a 50 lb. instrumented satellite as an end mass using a 20 km tether. Langley Research Center is providing the end mass instrumentation, while the Marshall Space Flight Center is designing and building the deployer. The objective of the experiment is to test the SEDS design concept by demonstrating that the system will satisfactorily deploy the full 20 km tether without stopping prematurely, come to a smooth stop on the application of a brake, and cut the tether at the proper time after it swings to the local vertical. Also, SEDS1 will collect data which will be used to test the accuracy of tether dynamics models used to stimulate this type of deployment. The experiment will last about 1.5 hours and complete approximately 1.5 orbits. Radar tracking of the Delta II and end mass is planned. In addition, the SEDS1 on-board computer will continuously record, store, and transmit mission data over the Delta II S-band telemetry system. The Data System will count tether windings as the tether unwinds, log the times of each turn and other mission events, monitor tether tension, and record the temperature of system components. A summary of the measurements taken during the SEDS1 are shown. The Data System will also control the tether brake and cutter mechanisms. Preliminary versions of two major sections of the flight software, the data telemetry modules and the data collection modules, were developed and tested under the 1990 NASA/ASEE Summer Faculty Fellowship Program. To facilitate the debugging of these software modules, a prototype SEDS Data System was programmed to simulate turn count signals. During the 1991 summer program, the concept of simulating signals produced by the SEDS electronics systems and circuits was expanded and more precisely defined. During the 1992 summer program, the SEDS signal simulator was programmed to test the requirements of the SEDS Mission software, and this simulator will be used in the formal verification of the SEDS Mission Software. The formal test procedures specification was written which incorporates the use of the signal simulator to test the SEDS Mission Software and which incorporates procedures for testing the other major component of the SEDS software, the Monitor Software.

Pierson, William E.↗

Vector wind profile gust model

To enable development of a vector wind gust model suitable for orbital flight test operations and trade studies, hypotheses concerning the distributions of gust component variables were verified. Methods for verification of hypotheses that observed gust variables, including gust component magnitude, gust length, u range, and L range, are gamma distributed and presented. Observed gust modulus has been drawn from a bivariate gamma distribution that can be approximated with a Weibull distribution. Zonal and meridional gust components are bivariate gamma distributed. An analytical method for testing for bivariate gamma distributed variables is presented. Two distributions for gust modulus are described and the results of extensive hypothesis testing of one of the distributions are presented. The validity of the gamma distribution for representation of gust component variables is established.

Adelfang, S. I.↗

GFO-1 Geophysical Data Record and Orbit Verifications for Global Change Studies

This final report summarizes the research work conducted under NASA's Physical Oceanography Program, entitled, GFO-1 Geophysical Data Record And Orbit Verifications For Global Change Studies, for the investigation time period from December 1, 1997 through November 30, 2000. The primary objectives of the investigation include providing verification and improvement for the precise orbit, media, geophysical, and instrument corrections to accurately reduce U.S. Navy's Geosat-Followon-1 (GFO-1) mission radar altimeter data to sea level measurements. The status of the GFO satellite (instrument and spacecraft operations, orbital tracking and altimeter) is summarized. GFO spacecraft has been accepted by the Navy from Ball Aerospace and has been declared operational since November, 2000. We have participated in four official GFO calibration/validation periods (Cal/Val I-IV), spanning from June 1999 through October 2000. Results of verification of the GFO orbit and geophysical data record measurements both from NOAA (IGDR) and from the Navy (NGDR) are reported. Our preliminary results indicate that: (1) the precise orbit (GSFC and OSU) can be determined to approx. 5 - 6 cm rms radially using SLR and altimeter crossovers; (2) estimated GFO MOE (GSFC or NRL) radial orbit accuracy is approx. 7 - 30 cm and Operational Doppler orbit accuracy is approx. 60 - 350 cm. After bias and tilt adjustment (1000 km arc), estimated Doppler orbit accuracy is approx. 1.2 - 6.5 cm rms and the MOE accuracy is approx. 1.0 - 2.3 cm; (3) the geophysical and media corrections have been validated versus in situ measurements and measurements from other operating altimeters (T/P and ERS-2). Altimeter time bias is insignificant with 0-2 ms. Sea state bias is about approx. 3 - 4.5% of SWH. Wet troposphere correction has approx. 1 cm bias and approx. 3 cm rms when compared with ERS-2 data. Use of GIM and IRI95 provide ionosphere correction accurate to 2-3 cm rms during medium to high solar activities; (4) the noise of the GFO altimeter data (uncorrected SSH) is about 15 mm, compared to 19 min for ERS-2, and 12 min for TOPEX. It is anticipated that the operational GFO-1 altimeter data will contribute to a number of researches in physical oceanography. A list of relevant presentations and publications is attached.

Shum, C. K.↗

Spectroscopic Determination of Trace Contaminants in High Purity Oxygen

Oxygen used for extravehicular activities (EVA) must be free of contaminants because a difference in a few tenths of a percent of argon or nitrogen content can mean significant reduction in available EVA time. These inert gases build up in the extravehicular mobility unit because they are not metabolized or scrubbed from the atmosphere. Measurement of oxygen purity above 99.5% is problematic, and currently only complex instruments such as gas chromatographs or mass spectrometers are used for these determinations. Because liquid oxygen boil-off from the space shuttle will no longer be available to supply oxygen for EVA use, other concepts are being developed to produce and validate high purity oxygen from cabin air aboard the International Space Station. A prototype optical emission technique capable of detecting argon and nitrogen below 0.1% in oxygen was developed at White Sands Test Facility. This instrument uses a glow discharge in reduced pressure gas to produce atomic emission from the species present. Because the atomic emission lines from oxygen, nitrogen, and argon are discrete and in many cases well-separated, trace amounts of argon and nitrogen can be detected in the ultraviolet and visible spectrum. This is a straightforward, direct measurement of the target contaminants and may lend itself to a device capable of on-orbit verification of oxygen purity. System design and optimized measurement parameters are presented.

Hornung, Steven D.↗

Software and Human-Machine Interface Development for Environmental Controls Subsystem Support

The Space Launch System (SLS) is the next premier launch vehicle for NASA. It is the next stage of manned space exploration from American soil, and will be the platform in which we push further beyond Earth orbit. In preparation of the SLS maiden voyage on Exploration Mission 1 (EM-1), the existing ground support architecture at Kennedy Space Center required significant overhaul and updating. A comprehensive upgrade of controls systems was necessary, including programmable logic controller software, as well as Launch Control Center (LCC) firing room and local launch pad displays for technician use. Environmental control acts as an integral component in these systems, being the foremost system for conditioning the pad and extremely sensitive launch vehicle until T-0. The Environmental Controls Subsystem (ECS) required testing and modification to meet the requirements of the designed system, as well as the human factors requirements of NASA software for Validation and Verification (V&V). This term saw significant strides in the progress and functionality of the human-machine interfaces used at the launch pad, and improved integration with the controller code.

Dobson, Matthew↗

NASA’s Developments in Cryogenic Fluid Management Technology

NASA has been pursuing the development and demonstration of enabling and enhancing technologies for Cryogenic Fluid Management (CFM) in collaboration with the US industry. NASA’s vision for current and future space exploration requires high-performance chemical and nuclear propulsion solutions that utilize cryogens. The Agency will use cryogenic fluids for Earth-to-orbit transportation, human missions to the Moon and Mars, planetary exploration, and in-situ resource utilization (ISRU) production. All these architectures will require state-of-the-art long-duration storage on the surface and in-space transfer for refueling. This paper focuses on key CFM technology developments completed or ongoing by the Cryogenic Fluid Management Portfolio Office within NASA’s Space Technology Mission Directorate. The CFM Portfolio Office, partnered with US aerospace industry, has invested in CFM technology ground and flight demonstrations, focused on maturing CFM hardware and operations for Artemis and Moon to Mars activities. These demonstrations additionally provide NASA with data to inform numerical modeling activities for verification and validation of design tools and approaches for future missions. This paper includes discussion on remaining high-priority open CFM technology gaps for Mar-forward architectures and initiatives for closure.

Cryogenic Fluid Management↗

The New NASA Orbital Debris Engineering Model ORDEM2000

The NASA Orbital Debris Program Office at Johnson Space Center has developed a new computer-based orbital debris engineering model, ORDEM2000, which describes the orbital debris environment in the low Earth orbit region between 200 and 2000 km altitude. The model is appropriate for those engineering solutions requiring knowledge and estimates of the orbital debris environment (debris spatial density, flux, etc.). ORDEM2000 can also be used as a benchmark for ground-based debris measurements and observations. We incorporated a large set of observational data, covering the object size range from 10 mm to 10 m, into the ORDEM2000 debris database, utilizing a maximum likelihood estimator to convert observations into debris population probability distribution functions. These functions then form the basis of debris populations. We developed a finite element model to process the debris populations to form the debris environment. A more capable input and output structure and a user-friendly graphical user interface are also implemented in the model. ORDEM2000 has been subjected to a significant verification and validation effort. This document describes ORDEM2000, which supersedes the previous model, ORDEM96. The availability of new sensor and in situ data, as well as new analytical techniques, has enabled the construction of this new model. Section 1 describes the general requirements and scope of an engineering model. Data analyses and the theoretical formulation of the model are described in Sections 2 and 3. Section 4 describes the verification and validation effort and the sensitivity and uncertainty analyses. Finally, Section 5 describes the graphical user interface, software installation, and test cases for the user.

Liou, Jer-Chyi↗

Real-Time Culture-Independent Microbial Profiling Onboard the International Space Station using Nanopore Sequencing

For the past two decades, microbial monitoring of the International Space Station (ISS) has relied on culture-dependent methods that require return to Earth for analysis. This has a number of limitations, with the most significant being bias towards the detection of culturable organisms and the inherent delay between sample collection and ground-based analysis. In recent years, portable and easy-to-use molecular-based tools, such as Oxford Nanopore Technologies’ MinION™ sequencer and miniPCR bio’s miniPCR™ thermal cycler, have been validated onboard the ISS. Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS. Method development focused on analysis of swabs collected in a low-biomass environment with limited facility resources and stringent controls on allowed processes and reagents. ISS-optimized procedures included enzymatic DNA extraction from a swab tip, bead-based purifications, altered buffers, and the use of miniPCR and the MinION. Validation was conducted through extensive ground-based assessments comparing current standard culture-dependent and newly developed culture-independent methods. Similar microbial distributions were observed between the two methods; however, as expected, the culture-independent data revealed microbial profiles with greater diversity. Protocol optimization and verification was established during NASA Extreme Environment Mission Operations (NEEMO) analog missions 21 and 22, respectively. Unique microbial profiles obtained from analog testing validated the swab-to-sequencer method in an extreme environment. Finally, four independent swab-to-sequencer experiments were conducted onboard the ISS by two crewmembers. Microorganisms identified from ISS swabs were consistent with historical culture-based data, and primarily consisted of commonly observed human-associated microbes. This simplified method has been streamlined for high ease-of-use for a non-trained crew to complete in an extreme environment, thereby enabling environmental and human health diagnostics in real-time as future missions take us beyond low-Earth orbit.

microbiology↗