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NASA NTRS2012

Fabrication and Testing of CERMET Fuel Materials for Nuclear Thermal Propulsion

A first generation Nuclear Cryogenic Propulsion Stage (NCPS) based on Nuclear Thermal Propulsion (NTP) is currently being developed for Advanced Space Exploration Systems. The overall goal of the project is to address critical NTP technology challenges and programmatic issues to establish confidence in the affordability and viability of NTP systems. The current technology roadmap for NTP identifies the development of a robust fuel form as a critical near term need. The lack of a qualified nuclear fuel is a significant technical risk that will require a considerable fraction of program resources to mitigate. Due to these risks and the cost for qualification, the development and selection of a primary fuel must begin prior to Authority to Proceed (ATP) for a specific mission. The fuel development is a progressive approach to incrementally reduce risk, converge the fuel materials, and mature the design and fabrication process of the fuel element. A key objective of the current project is to advance the maturity of CERMET fuels. The work includes fuel processing development and characterization, fuel specimen hot hydrogen screening, and prototypic fuel element testing. Early fuel materials development is critical to help validate requirements and fuel performance. The purpose of this paper is to provide an overview and status of the work at Marshall Space Flight Center (MSFC).

Hickman, Robert↗

NASA NTRS2012

Navigation Between Geosynchronous and Lunar L1 Orbiters

Linked Autonomous Interplanetary Satellite Orbit Navigation (LiAISON) is a new technique that takes advantage of the asymmetrical gravity field present in a three-body system in order to perform absolute tracking of satellites using only relative satellite-to-satellite observations. Previous studies have demonstrated LiAISON's practical applications for lunar missions, including a satellite in a halo orbit about either the Earth-Moon L1 or L2 point. This paper studies the viability of applying LiAISON measurements between a lunar halo orbiter and a satellite in a geosynchronous orbit. Simulations demonstrate that the absolute positions and velocities of both satellites are observable using only relative measurements with an achieved uncertainty on the order of observation noise.

low-energy↗

NASA NTRS2012

Extension of the Time-Spectral Approach to Overset Solvers for Arbitrary Motion

Forced periodic flows arise in a broad range of aerodynamic applications such as rotorcraft, turbomachinery, and flapping wing configurations. Standard practice involves solving the unsteady flow equations forward in time until the initial transient exits the domain and a statistically stationary flow is achieved. It is often required to simulate through several periods to remove the initial transient making unsteady design optimization prohibitively expensive for most realistic problems. An effort to reduce the computational cost of these calculations led to the development of the Harmonic Balance method [1, 2] which capitalizes on the periodic nature of the solution. The approach exploits the fact that forced temporally periodic flow, while varying in the time domain, is invariant in the frequency domain. Expanding the temporal variation at each spatial node into a Fourier series transforms the unsteady governing equations into a steady set of equations in integer harmonics that can be tackled with the acceleration techniques afforded to steady-state flow solvers. Other similar approaches, such as the Nonlinear Frequency Domain [3,4,5], Reduced Frequency [6] and Time-Spectral [7, 8, 9] methods, were developed shortly thereafter. Additionally, adjoint-based optimization techniques can be applied [10, 11] as well as frequency-adaptive methods [12, 13, 14] to provide even more flexibility to the method. The Fourier temporal basis functions imply spectral convergence as the number of harmonic modes, and correspondingly number of time samples, N, is increased. Some elect to solve the equations in the frequency domain directly, while others choose to transform the equations back into the time domain to simplify the process of adding this capability to existing solvers, but each harnesses the underlying steady solution in the frequency domain. These temporal projection methods will herein be collectively referred to as Time-Spectral methods. Time-Spectral methods have demonstrated marked success in reducing the computational costs associated with simulating periodic forced flows, but have yet to be fully applied to overset or Cartesian solvers for arbitrary motion with dynamic hole-cutting. Overset and Cartesian grid methodologies are versatile techniques capable of handling complex geometry configurations in practical engineering applications, and the combination of the Time-Spectral approach with this general capability potentially provides an enabling new design and analysis tool. In an arbitrary moving-body scenario for these approaches, a Lagrangian body moves through a fixed Eulerian mesh and mesh points in the Eulerian mesh interior to the solid body are removed (cut or blanked), leaving a hole in the Eulerian mesh. During the dynamic motion some gridpoints in the domain are blanked and do not have a complete set of time-samples preventing a direct implementation of the Time-Spectral method. Murman[6] demonstrated the Time-Spectral approach for a Cartesian solver with a rigid domain motion, wherein the hole cutting remains constant. Similarly, Custer et al. [15, 16] used the NASA overset OVERFLOW solver and limited the amount of relative motion to ensure static hole-cutting and interpolation. Recently, Mavriplis and Mundis[17] demonstrated a qualitative method for applying the Time-Spectral approach to an unstructured overset solver for arbitrary motion. The goal of the current work is to develop a robust and general method for handling arbitrary motion with the Time-Spectral approach within an overset or Cartesian mesh method, while still approaching the spectral convergence rate of the original Time-Spectral approach. The viscous OVERFLOW solver will be augmented with the new Time-Spectral algorithm and the capability of the method for benchmark problems in rotorcraft and turbomachinery will be demonstrated. This abstract begins with a brief synopsis of the Time-Spectral approach for overset grids and provides details of e current approach to allow for arbitrary motion. Model problem results in one and two dimensions are included to demonstrate the viability of the method and the convergence properties. Section IV briefly outlines the implementation into the OVERFLOW solver, and the abstract closes with a description of the benchmark test cases which will be included in the final paper.

Leffell, Joshua Isaac↗

NASA NTRS2012

Particle Filtering for Model-Based Anomaly Detection in Sensor Networks

A novel technique has been developed for anomaly detection of rocket engine test stand (RETS) data. The objective was to develop a system that postprocesses a csv file containing the sensor readings and activities (time-series) from a rocket engine test, and detects any anomalies that might have occurred during the test. The output consists of the names of the sensors that show anomalous behavior, and the start and end time of each anomaly. In order to reduce the involvement of domain experts significantly, several data-driven approaches have been proposed where models are automatically acquired from the data, thus bypassing the cost and effort of building system models. Many supervised learning methods can efficiently learn operational and fault models, given large amounts of both nominal and fault data. However, for domains such as RETS data, the amount of anomalous data that is actually available is relatively small, making most supervised learning methods rather ineffective, and in general met with limited success in anomaly detection. The fundamental problem with existing approaches is that they assume that the data are iid, i.e., independent and identically distributed, which is violated in typical RETS data. None of these techniques naturally exploit the temporal information inherent in time series data from the sensor networks. There are correlations among the sensor readings, not only at the same time, but also across time. However, these approaches have not explicitly identified and exploited such correlations. Given these limitations of model-free methods, there has been renewed interest in model-based methods, specifically graphical methods that explicitly reason temporally. The Gaussian Mixture Model (GMM) in a Linear Dynamic System approach assumes that the multi-dimensional test data is a mixture of multi-variate Gaussians, and fits a given number of Gaussian clusters with the help of the wellknown Expectation Maximization (EM) algorithm. The parameters thus learned are used for calculating the joint distribution of the observations. However, this GMM assumption is essentially an approximation and signals the potential viability of non-parametric density estimators. This is the key idea underlying the new approach.

Solano, Wanda↗

NASA NTRS2012

Development of Two Color Fluorescent Imager and Integrated Fluidic System for Nanosatellite Biology Applications

Nanosatellites offer frequent, low-cost space access as secondary payloads on launches of larger conventional satellites. We summarize the payload science and technology of the Microsatellite in-situ Space Technologies (MisST) nanosatellite for conducting automated biological experiments. The payload (two fused 10-cm cubes) includes 1) an integrated fluidics system that maintains organism viability and supports growth and 2) a fixed-focus imager with fluorescence and scattered-light imaging capabilities. The payload monitors temperature, pressure and relative humidity, and actively controls temperature. C. elegans (nematode, 50 m diameter x 1 mm long) was selected as a model organism due to previous space science experience, its completely sequenced genome, size, hardiness, and the variety of strains available. Three strains were chosen: two green GFP-tagged strains and one red tdTomato-tagged strain that label intestinal, nerve, and pharyngeal cells, respectively. The integrated fluidics system includes bioanalytical and reservoir modules. The former consists of four 150 L culture wells and a 4x5 mm imaging zone the latter includes two 8 mL fluid reservoirs for reagent and waste storage. The fluidic system is fabricated using multilayer polymer rapid prototyping: laser cutting, precision machining, die cutting, and pressure-sensitive adhesives it also includes eight solenoid-operated valves and one mini peristaltic pump. Young larval-state (L2) nematodes are loaded in C. elegans Maintenance Media (CeMM) in the bioanalytical module during pre-launch assembly. By the time orbit is established, the worms have grown to sufficient density to be imaged and are fed fresh CeMM. The strains are pumped sequentially into the imaging area, imaged, then pumped into waste. Reagent storage utilizes polymer bags under slight pressure to prevent bubble formation in wells or channels. The optical system images green and red fluorescence bands by excitation with blue (473 nm peak) and amber (587 nm peak) LEDs it achieves 8 m lateral resolution using a CMOS imaging chip (as configured for serial data speeds) or 4 m resolution using USB imaging chips. The imager consists of a modified commercial off-the-shelf CMOS chip camera, amber, blue and white LEDs, as well as a relay lens and dual-band filters to obviate moving parts while supporting both fluorescence wavelengths.

Wu, Diana Terri↗

NASA NTRS2013

Arcjet Testing of Woven Carbon Cloth for Use on Adaptive Deployable Entry Placement Technology

This paper describes arcjet testing and analysis that has successfully demonstrated the viability of three dimensional woven carbon cloth for dual use in the Adaptive Deployable Entry Placement Technology (ADEPT). ADEPT is an umbrella-like entry system that is folded for stowage in the launch vehicle s shroud and deployed in space prior to reaching the atmospheric interface. A key feature of the ADEPT concept is its lower ballistic coefficient for delivery of a given payload than those for conventional, rigid body entry systems. The benefits that accrue from the lower ballistic coefficient include factor of ten reductions of deceleration forces and entry heating. The former enables consideration of new classes of scientific instruments for solar system exploration while the latter enables the design of a more efficient thermal protection system. The carbon cloth now base lined for ADEPT has a dual use in that it serves as ADEPT s thermal protection system and as the "skin" that transfers aerodynamic deceleration loads to its umbrella-like substructure. The arcjet testing described in this paper was conducted for some of the higher heating conditions for a future Venus mission using the ADEPT concept, thereby showing that the carbon cloth can perform in a relevant entry environment. The ADEPT project considered the carbon cloth to be mission enabling and was carrying it as a major risk during Fiscal Year 2012. The testing and analysis reported here played a major role in retiring that risk and is highly significant to the success and possible adoption of ADEPT for future NASA missions. Finally, this paper also describes a preliminary engineering level code, based on the arcjet data, that can be used to estimate cloth thickness for future missions using ADEPT and to predict carbon cloth performance in future arcjet tests.

Arnold, James O.↗

NASA NTRS2013

Advances in the Development of a WCl6 CVD System for Coating UO2 Powders with Tungsten

Demonstrated viability and utilization of: a) Fluidized powder bed. b) WCl6 CVD process. c) Coated spherical particles with tungsten. The highly corrosive nature of the WCl6 solid reagent limits material of construction. Indications that identifying optimized process variables with require substantial effort and will likely vary with changes in fuel requirements.

Mireles, Omar R.↗

NASA NTRS2013

UV/Visible Telescope with Hubble Disposal

Submission Overview: Our primary objective is to convey a sense of the significant advances possible in astrophysics investigations for major Cosmic Origins COR program goals with a 2.4m telescope asset outfitted with one or more advanced UV visible instruments. Several compelling science objectives were identified based on community meetings these science objectives drove the conceptual design of instruments studied by the COR Program Office during July September 2012. This RFI submission encapsulates the results of that study, and suggests that a more detailed look into the instrument suite should be conducted to prove viability and affordability to support the demonstrated scientific value. This study was conducted in the context of a larger effort to consider the options available for a mission to dispose safely of Hubble hence, the overall architecture considered for the mission we studied for the 2.4m telescope asset included resource sharing. This mitigates combined cost and risk and provides naturally for a continued US leadership role in astrophysics with an advanced, general-purpose UV visible space telescope.

Benford, Dominic J.↗

NASA NTRS2008

Metrology - Beyond the Calibration Lab

We rely on data from measurements every day; a gas-pump, a speedometer, and a supermarket weight scale are just three examples of measurements we use to make decisions. We generally accept the data from these measurements as "valid." One reason we can accept the data is the "legal metrology" requirements established and regulated by the government in matters of commerce. The measurement data used by NASA, other government agencies, and industry can be critical to decisions which affect everything from economic viability, to mission success, to the security of the nation. Measurement data can even affect life and death decisions. Metrology requirements must adequately provide for risks associated with these decisions. To do this, metrology must be integrated into all aspects of an industry including research, design, testing, and product acceptance. Metrology, the science of measurement, has traditionally focused on the calibration of instruments, and although instrument calibration is vital, it is only a part of the process that assures quality in measurement data. For example, measurements made in research can influence the fundamental premises that establish the design parameters, which then flow down to the manufacturing processes, and eventually impact the final product. Because a breakdown can occur anywhere within this cycle, measurement quality assurance has to be integrated into every part of the life-cycle process starting with the basic research and ending with the final product inspection process. The purpose of this paper is to discuss the role of metrology in the various phases of a product's life-cycle. For simplicity, the cycle will be divided in four broad phases, with discussions centering on metrology within NASA. .

Mimbs, Scott M.↗

NASA NTRS2009

Optimization of Photovoltaic Performance Through the Integration of Electrodynamic Dust Shield Layers

The viability of photovoltaics on the Lunar and Martian surfaces may be determined by their ability to withstand significant degradation in the Lunar and Martian environments. One of the greatest threats is posed by fine dust particles which are continually blown about the surfaces. In an effort to determine the extent of the threat, and to investigate some abatement strategies, a series of experiments were conducted outdoors and in the Moon and Mars environmental chamber at the Florida Solar Energy Center. Electrodynamic dust shield prototypes based on the electric curtain concept have been developed by our collaborators at the Kennedy Space Center [1]. These thin film layers can remove dust from surfaces and prevent dust accumulation. Several types of dust shields were designed, built and tested under high vacuum conditions and simulated lunar gravity to validate the technology for lunar exploration applications. Gallium arsenide, single crystal and polycrystalline silicon photovoltaic integrated devices were designed, built and tested under Moon and Mars environmental conditions as well as under ambient conditions. Photovoltaic efficiency measurements were performed on each individual cell with the following configurations; without an encapsulation layer, with a glass covering, and with various thin film dust shields. It was found that the PV efficiency of the hybrid systems was unaffected by these various thin film dust shields, proving that the optical transmission of light through the device is virtually uninhibited by these layers. The future goal of this project is to incorporate a photovoltaic cell as the power source for the electrodynamic dust shield system, and experimentally show the effective removal of dust obstructing any light incident on the cell, thus insuring power production is maximized over time.

Nason, Steven↗

NASA NTRS2008

Application of Smart Solid State Sensor Technology in Aerospace Applications

Aerospace applications require a range of chemical sensing technologies to monitor conditions in both space vehicles and aircraft operations. One example is the monitoring of oxygen. For example, monitoring of ambient oxygen (O2) levels is critical to ensuring the health, safety, and performance of humans living and working in space. Oxygen sensors can also be incorporated in detection systems to determine if hazardous leaks are occurring in space propulsion systems and storage facilities. In aeronautic applications, O2 detection has been investigated for fuel tank monitoring. However, as noted elsewhere, O2 is not the only species of interest in aerospace applications with a wide range of species of interest being relevant to understand an environmental or vehicle condition. These include combustion products such as CO, HF, HCN, and HCl, which are related to both the presence of a fire and monitoring of post-fire clean-up operations. This paper discusses the development of an electrochemical cell platform based on a polymer electrolyte, NAFION, and a three-electrode configuration. The approach has been to mature this basic platform for a range of applications and to test this system, combined with "Lick and Stick" electronics, for its viability to monitor an environment related to astronaut crew health and safety applications with an understanding that a broad range of applications can be addressed with a core technology.

Hunter, Gary W.↗

NASA NTRS2013

Progress in Payload Separation Risk Mitigation for a Deployable Venus Heat Shield

A deployable decelerator known as the Adaptive Deployable Entry and Placement Technology (ADEPT) offers substantial science and mass savings for the Venus In Situ Explorer (VISE) mission. The lander and science payload must be separated from ADEPT during atmospheric entry. This paper presents a trade study of the separation system concept of operations and provides a conceptual design of the baseline: aft-separation with a subsonic parachute. Viability of the separation system depends on the vehicle's dynamic stability characteristics during deceleration from supersonic to subsonic speeds. A trajectory sensitivity study presented shows that pitch damping and Venusian winds drive stability prior to parachute deployment, while entry spin rate is not a driver of stability below Mach 5. Additionally, progress in free-flight CFD techniques capable of computing aerodynamic damping parameters is presented. Exploratory simulations of ADEPT at a constant speed of Mach number of 0.8 suggest the vehicle may have an oscillation limit cycle near 5 angle-of-attack. The proposed separation system conceptual design is thought to be viable.

Smith, Brandon P.↗

NASA NTRS2013

Demonstration of Detection and Ranging Using Solvable Chaos

Acoustic experiments demonstrate a novel approach to ranging and detection that exploits the properties of a solvable chaotic oscillator. This nonlinear oscillator includes an ordinary differential equation and a discrete switching condition. The chaotic waveform generated by this hybrid system is used as the transmitted waveform. The oscillator admits an exact analytic solution that can be written as the linear convolution of binary symbols and a single basis function. This linear representation enables coherent reception using a simple analog matched filter and without need for digital sampling or signal processing. An audio frequency implementation of the transmitter and receiver is described. Successful acoustic ranging measurements are presented to demonstrate the viability of the approach.

Corron, Ned J.↗

NASA NTRS2013

Summary of Activities for Health Monitoring of Composite Overwrapped Pressure Vessels

This three-year project (FY12-14) will design and demonstrate the ability of new Magnetic Stress Gages for the measurement of stresses on the inner diameter of a Composite Overwrapped Pressure Vessel overwrap. The sensors are being tested at White Sands Testing Facility (WSTF) where the results will be correlated with a known nondestructive technique acoustic emission. The gages will be produced utilizing Meandering Winding Magnetometer (MWM) and/or MWM array eddy current technology. The ultimate goal is to utilize this technology for the health monitoring of Composite Overwrapped Pressure Vessels for all future flight programs. The first full-scale pressurization test was performed at WSTF in June 2012. The goals of this test were to determine adaptations of the magnetic stress gauge instrumentation that would be necessary to allow multiple sensors to monitor the vessel's condition simultaneously and to determine how the sensor response changes with sensor selection and orientation. The second full scale pressurization test was performed at WSTF in August 2012. The goals of this test were to monitor the vessel's condition with multiple sensors simultaneously, to determine the viability of the multiplexing units (MUX) for the application, and to determine if the sensor responses in different orientations are repeatable. For both sets of tests the vessel was pressured up to 6,000 psi to simulate maximum operating pressure. Acoustic events were observed during the first pressurization cycle. This suggested that the extended storage period prior to use of this bottle led to a relaxation of the residual stresses imparted during auto-frettage. The pressurization tests successfully demonstrated the use of multiplexers with multiple MWM arrays to monitor a vessel. It was discovered that depending upon the sensor orientation, the frequencies, and the sense element, the MWM arrays can provide a variety of complementary information about the composite overwrapped pressure vessel load conditions. For example, low frequency measurements can be used to monitor the overwrap thickness and changes associated with pressure level. High frequency data is dominated by the properties of the overwrap, including the fiber orientations and lay-up of the layers.

Russell, Rick↗

NASA NTRS2011

Benefits of Matching Domain Structure for Planning Software: The Right Stuff

We investigated the role of domain structure in software design. We compared 2 planning applications, for a Mission Control group (International Space Station), and measured users speed and accuracy. Based on our needs analysis, we identified domain structure and used this to develop new prototype software that matched domain structure better than the legacy system. We took a high-fidelity analog of the natural task into the laboratory and found (large) periformance differences, favoring the system that matched domain structure. Our task design enabled us to attribute better periormance to better match of domain structure. We ran through the whole development cycle, in miniature, from needs analysis through design, development, and evaluation. Doing so enabled inferences not just about the particular systems compared, but also provided evidence for the viability of the design process (particularly needs analysis) that we are exploring.

Billman, Dorrit Owen↗

NASA NTRS2013

Comparison of Failure Modes in 2-D and 3-D Woven Carbon Phenolic Systems

NASA Ames Research Center is developing Woven Thermal Protection System (WTPS) materials as a new class of heatshields for entry vehicles (Stackpoole). Currently, there are few options for ablative entry heatshield materials, none of which is ideally suited to the planetary probe missions currently of interest to NASA. While carbon phenolic was successfully used for the missions Pioneer Venus and Galileo (to Jupiter), the heritage constituents are no longer available. An alternate carbon phenolic would need to be qualified for probe missions, which is most efficient at heat fluxes greater than those currently of interest. Additional TPS materials such as Avcoat and PICA are not sufficiently robust for the heat fluxes required. As a result, there is a large TPS gap between the materials efficient at very high conditions (carbon phenolic) and those that are effective at low-moderate conditions (all others). Development of 3D Woven TPS is intended to fill this gap, targeting mid-density weaves that could with withstand mid-range heat fluxes between 1100 W/sq cm and 8000 W/sq cm (Venkatapathy (2012). Preliminary experimental studies have been performed to show the feasibility of WTPS as a future mid-range TPS material. One study performed in the mARC Jet Facility at NASA Ames Research Center characterized the performance of a 3D Woven TPS sample and compared it to 2D carbon phenolic samples at ply angles of 0deg, 23.5deg, and 90deg. Each sample contained similar compositions of phenolic and carbon fiber volume fractions for experimental consistency. The goal of this study was to compare the performance of the TPS materials by evaluating resulting recession and failure modes. After exposing both samples to similar heat flux and pressure conditions, the 2D carbon phenolic laminate was shown to experience significant delamination between layers and further pocketing underneath separated layers. The 3D Woven TPS sample did not experience the delamination or pocketing failure modes because z-fibers in the through-thickness direction provided extra reinforcement to hold material layers together. Therefore, the benefit of using a 3D weave architecture was shown to alleviate failure modes experienced by a 2D laminate sample of similar material composition. In summary this poster reviews the thermal response performance comparisons drawn between a 3D Woven TPS sample and 2D Carbon Phenolic samples after performing rigorous heating experiments in the mARC facility at NASA Ames. Although the mARC Facility is still in its developmental stages, researchers expect similar trends in failure modes observed from large scale arc jet facilities. This work helps demonstrate the viability of 3D Woven TPSs as a new TPS option for future atmospheric entry missions.

Rossman, Grant A.↗

NASA NTRS2013

Postural Responses Following Space Flight and Ground Based Analogs

With the transition from the Shuttle program to the International Space Station (ISS), the opportunity to fly sensorimotor experiments in a weightless environment has become increasingly more difficult to obtain. As a result, more investigations have turned to ground-based analogs as a way of evaluating an experiment's viability. The two primary analogs available to most investigators are 6deg head down bed rest (HDBR) and dry immersion (DI). For the time being, HDBR investigations have been associated with studies conducted in the United States while the Russians and several other European Union states have concentrated their efforts on using DI as the space flight analog of choice. While either model may be viable for cardiovascular, bone and other system changes, vestibular and sensorimotor investigators have retained serious reservations of either analog's potential to serve as a replacement for a true weightless environment. These reservations have merit, but it is worthwhile to consider that not all changes associated with sensorimotor function during space flight are the result of top-down modifications, but may also be due to the lack, or change, of appropriate support surfaces applying force to the bottom of the feet. To this end we have compared quiet stance postural responses between short duration Space Shuttle flights, long duration ISS flights and HDBR of varying duration. Using these three platforms, representing different modifications of support we investigated postural ataxia using a quiet stance model. Quiet stance was obtained by asking the subjects to stand upright on a force plate, eyes open, arms at the side of the body for three min. From the force plate we obtained average sway velocity in two axes as well as length of line (stabilogram). These parameters were then related to EMG activity recorded from the medial gastrocnemius and lateral tibialis. It is significant to note that postural ataxia measured as quiet stance shows analogous changes between HDBR and space flight. Primary differences across short duration, long duration space flight and HDBR are related to the length of exposure associated with both space flight and HDBR.

Kofman, Igor S.↗

NASA NTRS2013

Detection of Only Viable Bacterial Spores Using a Live/Dead Indicator in Mixed Populations

This method uses a photoaffinity label that recognizes DNA and can be used to distinguish populations of bacterial cells from bacterial spores without the use of heat shocking during conventional culture, and live from dead bacterial spores using molecular-based methods. Biological validation of commercial sterility using traditional and alternative technologies remains challenging. Recovery of viable spores is cumbersome, as the process requires substantial incubation time, and the extended time to results limits the ability to quickly evaluate the efficacy of existing technologies. Nucleic acid amplification approaches such as PCR (polymerase chain reaction) have shown promise for improving time to detection for a wide range of applications. Recent real-time PCR methods are particularly promising, as these methods can be made at least semi-quantitative by correspondence to a standard curve. Nonetheless, PCR-based methods are rarely used for process validation, largely because the DNA from dead bacterial cells is highly stable and hence, DNA-based amplification methods fail to discriminate between live and inactivated microorganisms. Currently, no published method has been shown to effectively distinguish between live and dead bacterial spores. This technology uses a DNA binding photoaffinity label that can be used to distinguish between live and dead bacterial spores with detection limits ranging from 109 to 102 spores/mL. An environmental sample suspected of containing a mixture of live and dead vegetative cells and bacterial endospores is treated with a photoaffinity label. This step will eliminate any vegetative cells (live or dead) and dead endospores present in the sample. To further determine the bacterial spore viability, DNA is extracted from the spores and total population is quantified by real-time PCR. The current NASA standard assay takes 72 hours for results. Part of this procedure requires a heat shock step at 80 degC for 15 minutes before the sample can be plated. Using a photoaffinity label would remove this step from the current assay as the label readily penetrates both live and dead bacterial cells. Secondly, the photoaffinity label can only penetrate dead bacterial spores, leaving behind the viable spore population. This would allow for rapid bacterial spore detection in a matter of hours compared to the several days that it takes for the NASA standard assay.

Behar, Alberto E.↗