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At least 37 records · Page 2

Automated Counting of Particles To Quantify Cleanliness

A machine vision system, similar to systems used in microbiological laboratories to count cultured microbes, has been proposed for quantifying the cleanliness of nominally precisely cleaned hardware by counting residual contaminant particles. The system would include a microscope equipped with an electronic camera and circuitry to digitize the camera output, a personal computer programmed with machine-vision and interface software, and digital storage media. A filter pad, through which had been aspirated solvent from rinsing the hardware in question, would be placed on the microscope stage. A high-resolution image of the filter pad would be recorded. The computer would analyze the image and present a histogram of sizes of particles on the filter. On the basis of the histogram and a measure of the desired level of cleanliness, the hardware would be accepted or rejected. If the hardware were accepted, the image would be saved, along with other information, as a quality record. If the hardware were rejected, the histogram and ancillary information would be recorded for analysis of trends. The software would perceive particles that are too large or too numerous to meet a specified particle-distribution profile. Anomalous particles or fibrous material would be flagged for inspection.

Rhode, James↗

Dynamic cell culture system (7-IML-1)

This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.

Cogoli, Augusto↗

Human factors in spacecraft design

This paper describes some of the salient implications of evolving mission parameters for spacecraft design. Among the requirements for future spacecraft are new, higher standards of living, increased support of human productivity, and greater accommodation of physical and cultural variability. Design issues include volumetric allowances, architecture and layouts, closed life support systems, health maintenance systems, recreational facilities, automation, privacy, and decor. An understanding of behavioral responses to design elements is a precondition for critical design decisions. Human factors research results must be taken into account early in the course of the design process.

Harrison, Albert A.↗

Automating Microbial Directed Evolution For Bioengineering Applications

From a micro-biology perspective, directed evolution is a technique that uses controlled environmental pressures to select for a desired phenotype. Directed evolution has the distinct advantage over rational design of not needing extensive knowledge of the genome or pathways associated with a microorganism to induce phenotypes. However, there are currently limitations to the applicability of this technique including being time-consuming, error-prone, and dependent on existing assays that may lack selectivity for the given phenotype. The AADEC (Autonomous Adaptive Directed Evolution Chamber) system is a proof-of-concept instrument to automate and improve the technique such that directed evolution can be used more effectively as a general bioengineering tool. A series of tests using the automated system and comparable by-hand survival assay measurements have been carried out using UV-C radiation and Escherichia coli cultures in order to demonstrate the advantages of the AADEC versus traditional implementations of directed evolution such as random mutagenesis. AADEC uses UV-C exposure as both a source of environmental stress and mutagenesis, so in order to evaluate the UV-C tolerance obtained from the cultures, a manual UV-C exposure survival assay was developed alongside the device to compare the survival fractions at a fixed dosage. This survival assay involves exposing E. coli to UV-C radiation using a custom-designed exposure hood to control the flux and dose. Surviving cells are counted then transferred to the next iteration and so on for several iterations to calculate the survival fractions for each exposure iteration.This survival assay primarily serves as a baseline for the AADEC device, allowing quantification of the differences between the AADEC system over the manual approach. The primary data of comparison is survival fractions; this is obtained by optical density and plate counts in the manual assay and by optical density growth curve fits pre- and post-exposure in the automated case. This data can then be compiled to calculate trends over the iterations to characterize increasing UV-C resistance of the E.coli strains. The observed trends are statistically indistinguishable through several iterations from both sources.

Bioengineering↗

Robotics in a controlled, ecological life support system

Controlled, Ecological Life Support Systems (CELSS) that utilize plants to provide food, water and oxygen could consume considerable amounts of labor unless crop production, recovery and processing are automated. Robotic manipulators equipped with special end-effectors and programmed to perform the sensing and materials handling tasks would minimize the amount of astronaut labor required. The Human Rated Test Facility (HRTF) planned for Johnson Space Center could discover and demonstrate techniques of crop production which can be reliably integrated with machinery to minimize labor requirements. Before the physical components (shelves, lighting fixtures, etc.) can be selected, a systems analysis must be performed to determine which alternative processes should be followed and how the materials handling tasks should be automated. Given that the current procedures used to grow crops in a CELSS may not be the best methods to automate, then what are the alternatives? How may plants be grown, harvested, processed for food, and the inedible components recycled? What commercial technologies current exist? What research efforts are underway to develop new technologies which might satisfy the need for automation in a CELSS? The answers to these questions should prove enlightening and provide some of the information necessary to perform the systems analysis. The planting, culturing, gathering, threshing and separation, food processing, and recovery of inedible portions of wheat were studied. The basic biological and materials handling processes of each task are defined and discussed. Current practices at Johnson Space Center and other NASA centers are described and compared to common production practices in the plant production industry. Technologies currently being researched which might be applicable are identified and illustrated. Finally, based on this knowledge, several scenarios are proposed for automating the tasks for wheat.

Miles, Gaines E.↗

Technology for increased human productivity and safety on orbit

Technologies are addressed that can facilitate the efficient performance of station operations on the Space Station Freedom (SSF) and thereby optimize the utilization of SSF for scientific research. The dedication of SSF capabilities to scientific study and to the payload-user community is a key goal of the program. Robotics applications are discussed in terms of automating the processing of experiment materials on-orbit by transferring ampules to a furnace system or by handling plant-tissue cultures. Noncontact temperature measurement and medical support technology are considered important technologies for maximizing time for scientific purposes. Detailed examinations are conducted of other technologies including advanced data systems and furnace designs. The addition of the listed technologies can provide an environment in which scientific research is more efficient and accurate.

Ambrus, Judith↗

Chemically defined medium and Caenorhabditis elegans

BACKGROUND: C. elegans has been established as a powerful genetic system. Use of a chemically defined medium (C. elegans Maintenance Medium (CeMM)) now allows standardization and systematic manipulation of the nutrients that animals receive. Liquid cultivation allows automated culturing and experimentation and should be of use in large-scale growth and screening of animals. RESULTS: We find that CeMM is versatile and culturing is simple. CeMM can be used in a solid or liquid state, it can be stored unused for at least a year, unattended actively growing cultures may be maintained longer than with standard techniques, and standard C. elegans protocols work well with animals grown in defined medium. We also find that there are caveats to using defined medium. Animals in defined medium grow more slowly than on standard medium, appear to display adaptation to the defined medium, and display altered growth rates as they change the composition of the defined medium. CONCLUSIONS: As was suggested with the introduction of C. elegans as a potential genetic system, use of defined medium with C. elegans should prove a powerful tool.

Culture Media/chemistry↗

Novel Algae Technology to Utilize CO 2 for Value Added Products

The Helios-NRG technology for converting coal-based flue CO 2 into valuable products has been successfully validated through a series of rigorous test programs. The key component, the MSC (Multi Stage Continuous System), played a pivotal role in this process. The MSC's various stages were seamlessly integrated and controlled by a second-generation automated system, surpassing project goals in terms of both algal productivity and capture efficiency. Notably, the MSC's operational duration improved while maintaining healthy cultures. The integrated system was rigorously tested under real outdoor conditions with varying sunlight levels and template flue gas. The final MSC test, conducted at the NCCC, successfully met project objectives.

01 COAL, LIGNITE, AND PEAT↗

NASA Bioculture System: From Experiment Definition to Flight Payload

Starting in 2015, the NASA Bioculture System will be available to the science community to conduct cell biology and microbiology experiments on ISS. The Bioculture System carries ten environmentally independent Cassettes, which house the experiments. The closed loop fluids flow path subsystem in each Cassette provides a perfusion-based method for maintain specimen cultures in a shear-free environment by using a biochamber based on porous hollow fiber bioreactor technology. Each Cassette contains an incubator and separate insulated refrigerator compartment for storage of media, samples, nutrients and additives. The hardware is capable of fully automated or manual specimen culturing and processing, including in-flight experiment initiation, sampling and fixation, up to BSL-2 specimen culturing, and the ability to up to 10 independent cultures in parallel for statistical analysis. The incubation and culturing of specimens in the Bioculture System is a departure from standard laboratory culturing methods. Therefore, it is critical that the PI has an understanding the pre-flight test required for successfully using the Bioculture System to conduct an on-orbit experiment. Overall, the PI will conduct a series of ground tests to define flight experiment and on-orbit implementation requirements, verify biocompatibility, and determine base bioreactor conditions. The ground test processes for the utilization of the Bioculture System, from experiment selection to flight, will be reviewed. Also, pre-flight test schedules and use of COTS ground test equipment (CellMax and FiberCell systems) and the Bioculture System will be discussed.

Bioculture System↗

Knowledge-Based Intelligent Software Support of Cellular Adaptation to Microgravity Investigations

One of the most significant new opportunities that the Space Station affords cell biologists is the ability to do long-term cultivation of cells in the space environment. This facility is essential for investigations that are primarily focused on effects requiring a longer timeline of observation than that provided by the STS (Space Transportation System) platform. Such work requires both very strong laboratory skills to properly and quickly interact with the hardware hosting the culture and deep knowledge of the cell biology domain in order to optimally react to unanticipated scientific developments. Such work can be enabled by advanced automation techniques that have recently been used in the STS-based Spacelab, and that are being readied for the Space Station. In this paper, we describe the adaptation of PI-in-a-Box, the first interactive space science assistant system, to the study of the effects of space flight on cell cycle progression and proliferation.

Groleau, Nick↗

Tracking Metabolic Changes in Microbial Culture using Redox Measurements

During long-term space missions, microbial cultures accumulate the effects of low-dose radiation, microgravity, and other factors; altered growth and metabolic activity may occur before viability effects. This could affect functionality of bioreactors or other bio-enabled mission systems, as well as shed light on human health. Spaceflight microbiology studies beyond the low Earth orbit exposure afforded by the ISS have been limited. Nanosatellites offer an increasingly popular alternative for deep space missions. However, the communications delay requires biofluidic automation of a pre-defined experimental protocol, and the lack of sample return (reliance on sensors in flight) can significantly limit feasible investigations. Previous biological CubeSats (PharmaSat, O/OREOS, EcAMSat) have used alamarBlue, an off-the-shelf formulation of the redox indicator dye resazurin, to track metabolic activity, as will BioSentinel, the upcoming interplanetary microbiology experiment. A series of ground experiments (see abstracts by Liddell, Santa Maria, and A. Kim) were conducted using a microbial culture system outfitted with an electrochemical sensor array (electrical conductivity, pH, oxidation-reduction potential, and dissolved oxygen) with alamarBlue and the same strain of Saccharomyces cerevisiae as BioSentinel. By improving mapping of measured changes in alamarBlue kinetics to physicochemical changes, and ultimately to biological alterations such as shifted metabolic pathways, this work supplements data analyses from past missions and planning for future missions using alamarBlue to characterize space radiation effects. Initial results indicate that alamarBlue acts like a redox buffer; its presence significantly changes redox kinetics in otherwise identical cultures. The initial color change (blue resazurin reduced to red/pink resorufin) appears as a redox plateau. A second plateau, likely corresponding to the second color transition (resorufin to the colorless hydroresorufin), occurs at a lower redox value. The relationship to carbon source exhaustion, dissolved oxygen depletion, cell death, and measured redox potential is complex and still under study.

Tracking↗

A modular suite of hardware enabling spaceflight cell culture research

BioServe Space Technologies, a NASA Research Partnership Center (RPC), has developed and operated various middeck payloads launched on 23 shuttle missions since 1991 in support of commercial space biotechnology projects. Modular cell culture systems are contained within the Commercial Generic Bioprocessing Apparatus (CGBA) suite of flight-qualified hardware, compatible with Space Shuttle, SPACEHAB, Spacelab and International Space Station (ISS) EXPRESS Rack interfaces. As part of the CGBA family, the Isothermal Containment Module (ICM) incubator provides thermal control, data acquisition and experiment manipulation capabilities, including accelerometer launch detection for automated activation and thermal profiling for culture incubation and sample preservation. The ICM can accommodate up to 8 individually controlled temperature zones. Command and telemetry capabilities allow real-time downlink of data and video permitting remote payload operation and ground control synchronization. Individual cell culture experiments can be accommodated in a variety of devices ranging from 'microgravity test tubes' or standard 100 mm Petri dishes, to complex, fed-batch bioreactors with automated culture feeding, waste removal and multiple sample draws. Up to 3 levels of containment can be achieved for chemical fixative addition, and passive gas exchange can be provided through hydrophobic membranes. Many additional options exist for designing customized hardware depending on specific science requirements.

manned↗

Studying Microbial Adaptation in the Laboratory: Sensor & Control Upgrades for an Experimental Evolution Biofluidics System

Experimental evolution (EE) involves iteratively exposing a microbial community to specific stressors to study its response to changes in environment over time. EE work is commonly done manually in the laboratory, but, when there are many environmental variables to measure and adjust, it is highly labor intensive, prone to human error, and challenging to scale. Single-purpose automated continuous culturing chambers exist, but implement only limited stressor types. A more general-purpose design is desirable. The BeING Lab at Ames Research Center created the prototype Automated Adaptive Directed Evolution Chamber (AADEC) to address these problems, beginning with Escherichia coli tolerance of short-wave ultraviolet (UV-C) radiation and of temperature. In newer versions, AADEC monitors microbial activity and can adjust the UV-C and temperature levels automatically. An optical density measurement is used to determine how many cells are present in the growth medium—over time, this corresponds to how many survive and reproduce. Oxidation-reduction potential provides information on consumed metabolic energy, and pH and electrical conductivity on metabolic products. Dissolved oxygen content is used to determine aerobic vs anaerobic growth. A Raspberry Pi computer processes all this data to set the UV-C stressor level. AADEC’s auxiliary systems include peristaltic pumps to change media and agitation to counteract cell settling. These actuators can also act as additional stressors. With the Raspberry Pi monitoring sensors and adjusting actuators in real time, AADEC takes measurements and controls the environment much more accurately than can be done with a manual EE implementation. The third and latest AADEC iteration is the first to simplify design and usage with circuits on PCBs and the ability to pre-program experimental protocols. Still planned is expansion to a multi-well design for the study of varying cell cultures in parallel, which will enable researchers to retain and re-inoculate cultures exhibiting the desired trait most strongly while flushing out others. AADEC’s special capabilities make it a valuable tool for studying life under multiple stressors, enabling scientists to replicate changes in climate on microbes for study in a lab setting.

Microbial Adaptation↗

Evaluation of the Paratrend Multi-Analyte Sensor for Potential Utilization in Long-Duration Automated Cell Culture Monitoring

BACKGROUND: Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments. While several single-analyte sensors exist to measure culture health, a multi-analyte sensor would simplify the cell culture system. One such multi-analyte sensor, the Paratrend 7 manufactured by Diametrics Medical, consists of three optical fibers for measuring pH, dissolved carbon dioxide (pCO(2)), dissolved oxygen (pO(2)), and a thermocouple to measure temperature. The sensor bundle was designed for intra-vascular measurements in clinical settings, and can be used in bioreactors operated both on the ground and in NASA's Space Shuttle and International Space Station (ISS) experiments. METHODS: A Paratrend 7 sensor was placed at the outlet of a bioreactor inoculated with BHK-21 (baby hamster kidney) cells. The pH, pCO(2), pO(2), and temperature data were transferred continuously to an external computer. Cell culture medium, manually extracted from the bioreactor through a sampling port, was also assayed using a bench top blood gas analyzer (BGA). RESULTS: Two Paratrend 7 sensors were used over a single cell culture experiment (64 days). When compared to the manually obtained BGA samples, the sensor had good agreement for pH, pCO(2), and pO(2) with bias (and precision) 0.005(0.024), 8.0 mmHg (4.4 mmHg), and 11 mmHg (17 mmHg), respectively for the first two sensors. A third Paratrend sensor (operated for 141 days) had similar agreement (0.02+/-0.15 for pH, -4+/-8 mm Hg for pCO(2), and 24+/-18 mmHg for pO(2)). CONCLUSION: The resulting biases and precisions are com- parable to Paratrend sensor clinical results. Although the pO(2) differences may be acceptable for clinically relevant measurement ranges, the O(2) sensor in this bundle may not be reliable enough for the ranges of pO(2) in these cell culture studies without periodic calibration.

Validation Studies↗

EVA Wiki - Transforming Knowledge Management for EVA Flight Controllers and Instructors

The EVA Wiki was recently implemented as the primary knowledge database to retain critical knowledge and skills in the EVA Operations group at NASA's Johnson Space Center by ensuring that information is recorded in a common, easy to search repository. Prior to the EVA Wiki, information required for EVA flight controllers and instructors was scattered across different sources, including multiple file share directories, SharePoint, individual computers, and paper archives. Many documents were outdated, and data was often difficult to find and distribute. In 2011, a team recognized that these knowledge management problems could be solved by creating an EVA Wiki using MediaWiki, a free and open-source software developed by the Wikimedia Foundation. The EVA Wiki developed into an EVA-specific Wikipedia on an internal NASA server. While the technical implementation of the wiki had many challenges, one of the biggest hurdles came from a cultural shift. Like many enterprise organizations, the EVA Operations group was accustomed to hierarchical data structures and individually-owned documents. Instead of sorting files into various folders, the wiki searches content. Rather than having a single document owner, the wiki harmonized the efforts of many contributors and established an automated revision controlled system. As the group adapted to the wiki, the usefulness of this single portal for information became apparent. It transformed into a useful data mining tool for EVA flight controllers and instructors, as well as hundreds of others that support EVA. Program managers, engineers, astronauts, flight directors, and flight controllers in differing disciplines now have an easier-to-use, searchable system to find EVA data. This paper presents the benefits the EVA Wiki has brought to NASA's EVA community, as well as the cultural challenges it had to overcome.

Johnston, Stephanie S.↗

EVA Wiki - Transforming Knowledge Management for EVA Flight Controllers and Instructors

The EVA (Extravehicular Activity) Wiki was recently implemented as the primary knowledge database to retain critical knowledge and skills in the EVA Operations group at NASA's Johnson Space Center by ensuring that information is recorded in a common, searchable repository. Prior to the EVA Wiki, information required for EVA flight controllers and instructors was scattered across different sources, including multiple file share directories, SharePoint, individual computers, and paper archives. Many documents were outdated, and data was often difficult to find and distribute. In 2011, a team recognized that these knowledge management problems could be solved by creating an EVA Wiki using MediaWiki, a free and open-source software developed by the Wikimedia Foundation. The EVA Wiki developed into an EVA-specific Wikipedia on an internal NASA server. While the technical implementation of the wiki had many challenges, the one of the biggest hurdles came from a cultural shift. Like many enterprise organizations, the EVA Operations group was accustomed to hierarchical data structures and individually-owned documents. Instead of sorting files into various folders, the wiki searches content. Rather than having a single document owner, the wiki harmonized the efforts of many contributors and established an automated revision control system. As the group adapted to the wiki, the usefulness of this single portal for information became apparent. It transformed into a useful data mining tool for EVA flight controllers and instructors, and also for hundreds of other NASA and contract employees. Program managers, engineers, astronauts, flight directors, and flight controllers in differing disciplines now have an easier-to-use, searchable system to find EVA data. This paper presents the benefits the EVA Wiki has brought to NASA's EVA community, as well as the cultural challenges it had to overcome.

Johnston, Stephanie↗

EVA Wiki - Transforming Knowledge Management for EVA Flight Controllers and Instructors

The EVA Wiki was recently implemented as the primary knowledge database to retain critical knowledge and skills in the EVA Operations group at NASA's Johnson Space Center by ensuring that information is recorded in a common, easy to search repository. Prior to the EVA Wiki, information required for EVA flight controllers and instructors was scattered across different sources, including multiple file share directories, SharePoint, individual computers, and paper archives. Many documents were outdated, and data was often difficult to find and distribute. In 2011, a team recognized that these knowledge management problems could be solved by creating an EVA Wiki using MediaWiki, a free and open-source software developed by the Wikimedia Foundation. The EVA Wiki developed into an EVA-specific Wikipedia on an internal NASA server. While the technical implementation of the wiki had many challenges, one of the biggest hurdles came from a cultural shift. Like many enterprise organizations, the EVA Operations group was accustomed to hierarchical data structures and individually-owned documents. Instead of sorting files into various folders, the wiki searches content. Rather than having a single document owner, the wiki harmonized the efforts of many contributors and established an automated revision controlled system. As the group adapted to the wiki, the usefulness of this single portal for information became apparent. It transformed into a useful data mining tool for EVA flight controllers and instructors, as well as hundreds of others that support the EVA. Program managers, engineers, astronauts, flight directors, and flight controllers in differing disciplines now have an easier-to-use, searchable system to find EVA data. This paper presents the benefits the EVA Wiki has brought to NASA's EVA community, as well as the cultural challenges it had to overcome.

Johnston, Stephanie S.↗

Comparative performance and technoeconomic analyses of two microalgae harvesting systems evaluated at a commercially relevant scale

Using a commercially scalable system designed for processing thousands of liters a day, here we evaluated the factors that affected the performance, energy consumption and capital/operating costs of both flocculation-based and filtration-based algal harvesting systems over a 16 month period. Coagulation efficiency was the primary driver of harvest efficiency in the flocculation-based method, while cell lysis was important for the filtration-based method. Culture (algae) age differentially influenced harvest efficiency through changes in cell fragility and stickiness. The average energy consumption of the flocculation-based method was 0.389 kWh/m 3 , while that for the filtration-based method was 4.343 kWh/m 3 . The average harvesting cost of the filtration-based method was 5.35 $\$$/m 3 , while for flocculation-based method it was 4.52 $\$$/m 3 . The concentration factor of filtration-based and flocculation-based method were 770–1086 and 407–448, respectively. For both harvesting methods, labor costs dominated and ranged from 55.84%–67.94% of total cost. Further system automation is a potential method to lower the harvesting cost. The filtration-based harvesting method could produce a better quality of algal biomass with higher concentration factors and less ash content, but needed more energy input, as compared with a flocculation-based method. This study highlights the importance of algal culture status to successful harvesting, and it also provides insight into developing more efficient harvesting technology with lower energy and capital cost.

59 BASIC BIOLOGICAL SCIENCES↗