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NASA NTRS2005

Medical and Scientific Evaluations aboard the KC-135. Microgravity-Compatible Flow Cytometer

A spaceflight-compatible flow cytometer would be useful for the diagnosis of astronaut illness during long duration spaceflight and for conducting in-flight research to evaluate the effects of microgravity on human physiology. Until recently, the primary limitations preventing the development of a spaceflight compatible flow cytometer have been largely mechanical. Standard commercially available flow cytometers are large, complex instruments that use high-energy lasers and require significant training to operate. Standard flow cytometers function by suspending the particles to be analyzed inside a sheath fluid for analysis. This requires the presence of several liters of sheath fluid for operation, and generates a corresponding amount of liquid hazardous waste. The particles are then passed through a flow cell which uses the fluid mechanical property of hydrodynamic focusing to place the cells in single-file (laminar flow) as they pass through a laser beam for scanning and evaluation. Many spaceflight experiments have demonstrated that fluid physics is dramatically altered in microgravity (MSF [Manned Space Flight] Fluid Physics Data Sheet-August 1997) and previous studies have shown that sheath-fluid based hydrodynamic focusing may also be altered during microgravity (Crucian et al, 2000). For these reasons it is likely that any spaceflight compatible design for a flow cytometer would abandon the sheath fluid requirement. The elimination of sheath fluid would remove both the problems of weight associated with large volumes of liquids as well as the large volume of liquid waste generated. It would also create the need for a method to create laminar particle flow distinct from the standard sheath-fluid based method. The spaceflight prototype instrument is based on a recently developed commercial flow cytometer possessing a novel flow cell design that creates single-particle laser scanning and evaluation without the need for sheath-fluid based hydrodynamic focusing. This instrument also possesses a number of design advances that make it conditionally microgravity compatible: it is highly miniaturized and lightweight, uses a low energy diode laser, has a small number of moving parts, does not use sheath fluid and does not generate significant liquid waste. Although possessing certain limitations, the commercial cytometer functions operationally like a standard bench top laboratory flow cytometer, aspirating liquid particle samples and generating histogram or dot-plot data in standard FCS file format. In its current configuration however, the cytometer is limited to three parameter/two-color capability (two color PMTs + forward scatter), does not allow compensation between colors, does not allow linear analysis and is operated by rather inflexible software with limited capabilities. This is due to the fact that the cytometer has been designed and marketed as an instrument specific to a few particular assays, not as a multipurpose cytometer.

Crucian, Brian↗

NASA NTRS1999

Detection of endogenous alkaline phosphatase activity in intact cells by flow cytometry using the fluorogenic ELF-97 phosphatase substrate

BACKGROUND: The alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (ELF((R))-97 for enzyme-labeled fluorescence) has been found useful for the histochemical detection of endogenous AP activity and AP-tagged proteins and oligonucleotide probes. In this study, we evaluated its effectiveness at detecting endogenous AP activity by flow cytometry. METHODS: The ELF-97 phosphatase substrate was used to detect endogenous AP activity in UMR-106 rat osteosarcoma cells and primary cultures of chick chondrocytes. Cells were labeled with the ELF-97 reagent and analyzed by flow cytometry using an argon ultraviolet (UV) laser. For comparison purposes, cells were also assayed for AP using a Fast Red Violet LB azo dye assay previously described for use in detecting AP activity by flow cytometry. RESULTS: The ELF-97 phosphatase substrate effectively detected endogenous AP activity in UMR-106 cells, with over 95% of the resulting fluorescent signal resulting from AP-specific activity (as determined by levamisole inhibition of AP activity). In contrast, less than 70% of the fluorescent signal from the Fast Red Violet LB (FRV) assay was AP-dependent, reflecting the high intrinsic fluorescence of the unreacted components. The ELF-97 phosphatase assay was also able to detect very low AP activity in chick chondrocytes that was undetectable by the azo dye method. CONCLUSIONS: The ELF-97 phosphatase assay was able to detect endogenous AP activity in fixed mammalian and avian cells by flow cytometry with superior sensitivity to previously described assays. This work also shows the applicability of ELF-97 to flow cytometry, supplementing its previously demonstrated histochemical applications. Copyright 1999 Wiley-Liss, Inc.

Non-NASA Center↗

NASA NTRS1975

Cost and size estimates for an electrochemical bulk energy storage concept

Preliminary capital cost and size estimates were made for an electrochemical bulk energy storage concept for a redox-flow-cell system. Preliminary calculations showed that the redox-flow-cell system has great promise as a bulk energy storage system for power load leveling. The size of the system was estimated to be less than 2 percent of the size of a comparable pumped hydroelectric storage plant.

Warshay, M.↗

NASA NTRS1985

A cluster analysis method for identification of subpopulations of cells in flow cytometric list-mode arrays

A specialized program was developed for flow cytometric list-mode data using an heirarchical tree method for identifying and enumerating individual subpopulations, the method of principal components for a two-dimensional display of 6-parameter data array, and a standard sorting algorithm for characterizing subpopulations. The program was tested against a published data set subjected to cluster analysis and experimental data sets from controlled flow cytometry experiments using a Coulter Electronics EPICS V Cell Sorter. A version of the program in compiled BASIC is usable on a 16-bit microcomputer with the MS-DOS operating system. It is specialized for 6 parameters and up to 20,000 cells. Its two-dimensional display of Euclidean distances reveals clusters clearly, as does its 1-dimensional display. The identified subpopulations can, in suitable experiments, be related to functional subpopulations of cells.

Li, Z. K.↗

NASA NTRS1974

Rheological properties of RBC in the microcirculation of mammalian skeletal muscle

In the investigation the established technique of direct microscopic viewing was combined with the use of a closed circuit television system and cinematography. The red cell flow patterns in all capillaries were found to be oscillatory with characteristic cycle frequencies and amplitudes for all concentrations of inspired oxygen greater than 8%. Generally, there was a transient decrease in mean flow rate with increasing severity of hypoxia, with a gradual return toward control values. Red cell flow patterns are discussed along with questions of red cell configuration.

Ehrenberg, M. H.↗

NASA NTRS1974

Methods for improved resolution of flow electrophoresis cells

First method involves remote adjusting of zeta potential. Second approach sandwiches two conducting metal plates between opposite cell walls and thin insulating layer. Third method forces buffer to flow in direction opposite particle streams.

Mccreight, L. R.↗

NASA NTRS1992

Effect of radiation on convection at moderate temperatures

A combined numerical and experimental investigation of radiation-induced convection is presented to show that the convective stability of the top-heated enclosure is disrupted by heat transfer conditions at the wall. When the enclosure is not insulated the thermal stratification of the fluid is modified by convective and radiative losses to the surrounding environment. This results in a double annular cell flow which, when cut by the laser sheet, shows a four-vortex pattern with a weak annular cell at the bottom and a large counter-rotating annular cell at the top. When the enclosure is insulated the convective stability of the fluid is again disrupted - this time as a result of radiative heat transfer between the enclosing surfaces which drives two annular flow cells of relatively equal size. Comparison between model and experiment shows that radiation effects are important even at temperature levels as low as 300 C and, if these effects are not included, numerical predictions can be highly erroneous.

Degroh, Henry C., III↗

NASA NTRS1992

Effect of radiation on convection at moderate temperatures

De Groh, H. C., III↗

NASA NTRS1975

Cost and size estimates for an electrochemical bulk energy storage concept

Preliminary capital cost and size estimates were made for an electrochemical bulk energy storage concept. The electrochemical system considered was an electrically rechargeable flow cell with a redox couple. On the basis of preliminary capital cost estimates, size estimates, and several other important considerations, the redox-flow-cell system emerges as having great promise as a bulk energy storage system for power load leveling. The size of this system would be less than 2 percent of that of a comparable pumped hydroelectric plant. The capital cost of a 10-megawatt, 60- and 85-megawatt-hour redox system is estimated to be $190 to $330 per kilowatt. The other important features of the redox system contributing to its load leveling application are its low adverse environmental impact, its high efficiency, its apparent absence of electrochemically-related cycle life limitations, and its fast response.

Warshay, M.↗

NASA NTRS2010

Electrochemical Detection of Multiple Bioprocess Analytes

An apparatus that includes highly miniaturized thin-film electrochemical sensor array has been demonstrated as a prototype of instruments for simultaneous detection of multiple substances of interest (analytes) and measurement of acidity or alkalinity in bioprocess streams. Measurements of pH and of concentrations of nutrients and wastes in cell-culture media, made by use of these instruments, are to be used as feedback for optimizing the growth of cells or the production of desired substances by the cultured cells. The apparatus is designed to utilize samples of minimal volume so as to minimize any perturbation of monitored processes. The apparatus can function in a potentiometric mode (for measuring pH), an amperometric mode (detecting analytes via oxidation/reduction reactions), or both. The sensor array is planar and includes multiple thin-film microelectrodes covered with hydrous iridium oxide. The oxide layer on each electrode serves as both a protective and electrochemical transducing layer. In its transducing role, the oxide provides electrical conductivity for amperometric measurement or pH response for potentiometric measurement. The oxide on an electrode can also serve as a matrix for one or more enzymes that render the electrode sensitive to a specific analyte. In addition to transducing electrodes, the array includes electrodes for potential control. The array can be fabricated by techniques familiar to the microelectronics industry. The sensor array is housed in a thin-film liquid-flow cell that has a total volume of about 100 mL. The flow cell is connected to a computer-controlled subsystem that periodically draws samples from the bioprocess stream to be monitored. Before entering the cell, each 100-mL sample is subjected to tangential-flow filtration to remove particles. In the present version of the apparatus, the electrodes are operated under control by a potentiostat and are used to simultaneously measure the pH and the concentration of glucose. It is anticipated that development of procedures for trapping more enzymes into hydrous iridium oxide (and possibly into other electroactive metal oxides) and of means for imparting long-term stability to the transducer layers should make it possible to monitor concentrations of products of many enzyme reactions for example, such key bioprocess analytes as amino acids, vitamins, lactose, and acetate.

Rauh, R. David↗

NASA NTRS1981

Design and assembly considerations for Redox cells and stacks

Individual redox flow cells are arranged electrically in series and hydraulically in parallel to form a single assembly called a stack. The hardware currently being tested in the laboratory has an active electrode area of either 310 sq cm or 929 sq cm. Four 310 sq cm stacks, each consisting of 39 active cells, were incorporated into a 1.0 kW preprototype system. The physical design of the stack is very critical to the performance and efficiency of the redox storage sytem. This report will discuss the mechanical aspects of the cell and stack design for the current Redox hardware, with regard to sealing the stack internally as well as externally, minimizing shunt currents and minimizing the electrical resistance of the stack.

Stalnaker, D. K.↗

NASA NTRS1994

Continuous monitoring of bacterial attachment

A major concern with the Space Station Freedom (SSF) water supply system is the control of longterm microbial contamination and biofilm development in the water storage and distribution systems. These biofilms have the potential for harboring pathogens as well as microbial strains containing resistance factors that could negatively influence crew health. The proposed means for disinfecting the water system on SSF (iodine) may encourage the selection of resistant strains. In fact, biofilm bacteria were observed in water lines from the Space Shuttle Columbia (OV-102); therefore, an alternative remediation method is required to disinfect spacecraft water lines. A thorough understanding of colonization events and the physiological parameters that will influence bacteria adhesion is required. The limiting factor for development of this technology is the ability to continuously monitor adhesion events and the effects of biocides on sessile bacteria. Methods were developed to allow bacterial adhesion and subsequent biocidal treatment to be monitored continuously. This technique couples automated image analysis with a continuous flow of a bacterial suspension through an optical flow cell. A strain of Pseudomonas cepacia isolated from the water supply of the Space Shuttle Discovery (OV-103) during STS-39 was grown in a nitrogen-limited continuous culture. This culture was challenged continuously with iodine during growth, and the adhesion characteristics of this strain was measure with regard to flow rate. Various biocides (ozone, hypochlorite, and iodine) were added to the flow stream to evaluate how well each chemical removed the bacteria. After biocide treatment, a fresh bacterial suspension was introduced into the flow cell, and the attachment rate was evaluated on the previously treated surface. This secondary fouling was again treated with biocide to determine the efficacy of multiple batch chemical treatments in removing biofilm.

Koeing, D. W.↗

NASA NTRS1996

Simulated 'On-Line' Wear Metal Analysis of Lubricating Oils by X-Ray Fluorescence Spectroscopy

The objective of this project was to assess the sensitivity of X-ray Fluorescence Spectroscopy (XFS) for quantitative evaluation of metal particle content in engine oil suspensions and the feasibility of real-time, dynamic wear metal analysis. The study was focused on iron as the majority wear metal component. Variable parameters were: particle size, particle concentration and oil velocity. A commercial XFS spectrometer equipped with interchangeable static/dynamic (flow cell) sample chambers was used. XFS spectra were recorded for solutions of Fe-organometallic standard and for a series of DTE oil suspensions of high purity spherical iron particles of 2g, 4g, and 8g diameter, at concentrations from 5 ppm to 5,000 ppm. Real contaminated oil samples from Langley Air Force Base aircraft engines and NASA Langley Research Center wind tunnels were also analyzed. The experimental data conform the reliability of XFS as the analytical method of choice for this project. Intrinsic inadequacies of the instrument for precise analytic work at low metal concentrations were identified as being related to the particular x-ray beam definition, system geometry, and flow-cell materials selection. This work supports a proposal for the design, construction and testing of a conceptually new, miniature XFS spectrometer with superior performance, dedicated to on-line, real-time monitoring of lubricating oils in operating engines. Innovative design solutions include focalization of the incident x-ray beam, non-metal sample chamber, and miniaturization of the overall assembly. The instrument would contribute to prevention of catastrophic engine failures. A proposal for two-year funding has been presented to NASA Langley Research Center Internal Operation Group (IOG) Management, to continue the effort begun by this summer's project.

Kelliher, Warren C.↗

NASA NTRS2013

New Class of Flow Batteries for Terrestrial and Aerospace Energy Storage Applications

Future sustainable energy generation technologies such as photovoltaic and wind farms require advanced energy storage systems on a massive scale to make the alternate (green) energy options practical. The daunting requirements of such large-scale energy systems such as long operating and cycle life, safety, and low cost are not adequately met by state-of-the-art energy storage technologies such as vanadium flow cells, lead-acid, and zinc-bromine batteries. Much attention is being paid to redox batteries specifically to the vanadium redox battery (VRB) due to their simplicity, low cost, and good life characteristics compared to other related battery technologies. NASA is currently seeking high-specific- energy and long-cycle-life rechargeable batteries in the 10-to-100-kW range to support future human exploration missions, such as planetary habitats, human rovers, etc. The flow batteries described above are excellent candidates for these applications, as well as other applications that propose to use regenerative fuel cells. A new flow cell technology is proposed based on coupling two novel electrodes in the form of solvated electron systems (SES) between an alkali (or alkaline earth) metal and poly aromatic hydrocarbons (PAH), separated by an ionically conducting separator. The cell reaction involves the formation of such SES with a PAH of high voltage in the cathode, while the alkali (or alkaline earth metal) is reduced from such an MPAH complex in the anode half-cell. During recharge, the reactions are reversed in both electrodes. In other words, the alkali (alkaline earth) metal ion simply shuttles from one M-PAH complex (SES) to another, which are separated by a metal-ion conducting solid or polymer electrolyte separator. As an example, the concept was demonstrated with Li-naphthalene//Li DDQ (DDQ is 2,3-Dichloro-5,6-dicyano- 1,4-benzoquinone) separated by lithium super ion conductor, either ceramic or polymer (solid polymer or gel polymer) electrolytes. The reactants are Li-naphthalene dissolved in tetrahydrofuran (THF) with a lithium salt of 1M LiBF4 (lithium tetra fluoroborate) in the anode compartment, and DDQ again dissolved in THF and also containing 1M LiBF4 salt in the cathode half-cell. The solid electrolyte separator used in the first set of experiments is a ceramic solid electrolyte, available from a commercial source. The open circuit voltage of the cells is close to 3.0 V, as expected from the individual half-cell voltages of Li-naphthalene and Li-DDQ. Upon discharge, the cell shows steady discharge voltage of 2.7 V, which confirms that the electrochemical processes do involve lithium ion shuttling from the anodic compartment to the cathode half-cell. The reversibility or rechargeability is demonstrated by charging the partially discharged cells (i.e., with lithium present in the DDQ half). Once again, a steady voltage close to 3.0 V was observed during charge, indicating that the system is quite reversible. In the subsequent concept-demonstration studies, the ceramic electrolyte has been replaced with a gel polymer electrolyte, e.g., PVDF-HFP (poly vinylene difluoride hexafluoropropene) gel, which has several advantages such as high ionic conductivity (almost comparable to liquid electrolyte and about 2 orders of magnitude better than the ceramic equivalent), lower cost, and possibly higher chemical stability at the anode. In addition, it can be bonded to the electrode by thermal fusion to form membrane electrode assemblies (MEAs), as is done in fuel cells.

Bugga, Ratnakumar V.↗

NASA NTRS1985

Urokinase production by electrophoretically separated cultured human embryonic kidney cells

Urokinase is a plasminogen activator found in urine. Relatively pure preparations have been tested in Europe, Japan and the United States for the treatment of deep vein thrombosis and other dangerous blood clots. Human embryonic kidney cell cultures have been found to produce urokinase at much higher concentrations, but less than 5% of the cells in typical cultures are producers. Since human diploid cells become senescent in culture the selection of clones derived from single cells will not provide enough material to be useful, so a bulk purification method is needed for the isolation of urokinase producing cell populations. Preparative cell electrophoresis was chosen as the method, since evidence exists that human embryonic cell cultures are richly heterogeneous with respect to electrophoretic mobility, and preliminary electrophoretic separations on the Apollo-Soyuz space flight produced cell populations that were rich in urokinase production. Similarly, erythropoietin is useful in the treatment of certain anemias and is a kidney cell duct, and electrophoretically enriched cell populations producing this product have been reported. Thus, there is a clear need for diploid human cells that produce these products, and there is evidence that such cells should be separable by free-flow cell electrophoresis.

Kunze, M. E.↗

NASA NTRS1988

On the relation between photospheric flow fields and the magnetic field distribution on the solar surface

Using the technique of local correlation tracking on a 28 minute time sequence of white-light images of solar granulation, the horizontal flow field on the solar surface is measured. The time series was obtained by the Solar Optical Universal Polarimeter (SOUP) on Spacelab 2 (Space Shuttle flight 51-F) and is free from atmospheric blurring and distortion. The SOUP flow fields have been compared with carefully aligned magnetograms taken over a nine hour period at the Big Bear Solar Observatory before, during, and after the SOUP images. The flow field and the magnetic field agree in considerable detail: vectors which define the flow of the white-light intensity pattern (granulation) point toward magnetic field regions, magnetic fields surround flow cells, and magnetic features move along the flow arrows. The projected locations of free particles ('corks') in the measured flow field congregate at the same locations where the magnetic field is observed.

Simon, George W.↗

NASA NTRS2011

The Advantages of Non-Flow-Through Fuel Cell Power Systems for Aerospace Applications

NASA has been developing proton-exchange-membrane (PEM) fuel cell power systems for the past decade, as an upgraded technology to the alkaline fuel cells which presently provide power for the Shuttle Orbiter. All fuel cell power systems consist of one or more fuel cell stacks in combination with appropriate balance-of-plant hardware. Traditional PEM fuel cells are characterized as flow-through, in which recirculating reactant streams remove product water from the fuel cell stack. NASA recently embarked on the development of non-flow-through fuel cell systems, in which reactants are dead-ended into the fuel cell stack and product water is removed by internal wicks. This simplifies the fuel cell power system by eliminating the need for pumps to provide reactant circulation, and mechanical water separators to remove the product water from the recirculating reactant streams. By eliminating these mechanical components, the resulting fuel cell power system has lower mass, volume, and parasitic power requirements, along with higher reliability and longer life. These improved non-flow-through fuel cell power systems therefore offer significant advantages for many aerospace applications.

Hoberecht, Mark↗

NASA NTRS2012

Non-Flow-Through Fuel Cell System Test Results and Demonstration on the SCARAB Rover

This presentation describes the results of the demonstration of a non-flow-through PEM fuel cell as part of a power system on the SCARAB rover at the NASA Glenn Research Center. A 16-cell non-flow-through fuel cell stack from Infinity Fuel Cell and Hydrogen, Inc. was incorporated into a power system designed to act as a range extender by providing power to the SCARAB rover s hotel loads. The power system, including the non-flow-through fuel cell technology, successfully demonstrated its goal as a range extender by powering hotel loads on the SCARAB rover, making this demonstration the first to use the non-flow-through fuel cell technology on a mobile platform.

Scheidegger, Brianne↗