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At least 73 records · Page 4

Advancing Launch Readiness of the Europa Light Microscope

The search for life in our solar system relies on our ability to measure key biomarkers in situ and identify them as likely of biotic origin. One class of biomarkers that would be indicative of life is cell-like features such as the colocalization of key biochemicals, spatial organization of native UV fluorescent compounds, and polarization across membrane compartments. The Europa Lander Science Definition Team has emphasized the need for a light microscope to search for these features on Europa, and the Europa Light Microscope (ELM) has been developed to address this need. This work presents the development towards high launch readiness of ELM by demonstrating its optical resolution under brightfield illumination, deep UV fluorescence imaging of unstained living samples, and multicolor imaging of samples stained with exogenous fluorophores targeting proteins, lipids, and nucleic acids. This work also seeks to extend the capabilities of ELM by incorporating fluorescence-based assays for membrane potential as a key biomarker for living cellular material.

Pavel Erich Zivanovic Klier↗

Spectrum

Spectrum is a multispectral fluorescence imager designed for capturing in vivo genetic expression in a variety of biological organisms, providing a capability that does not currently exist on the International Space Station (ISS). Researching organisms that have been transformed with in vivo reporter genes ligated with fluorescent proteins allows the scientific community to further understand the fundamental biological responses of these organisms when subjected to space environments. Model organisms that may utilize multispectral imaging on the ISS include unicellular organisms (e.g. Saccharomyces cerevisiae), plants (e.g. Arabidopsis thaliana), and invertebrates (e.g. Caenorhabditis elegans).

Levine, Howard G.↗

Design and Development of Modules to Support Live Microscopic Imaging on Ground-Based Microgravity Simulators

In space, astronauts are exposed to environmental stressors that often result in physiological changes. One prominent stressor in spaceflight is microgravity, and research has shown that long term microgravity exposure causes muscle atrophy, bone loss, cardiovascular concerns, and vision impairment. It is critical to understand how altered gravity affects physiology on the cellular, molecular, and gene level in order to accurately assess health risks and to develop effective countermeasures. Ground-based microgravity simulators such as random positioning machines (RPMs) are used to produce some of the biological effects of altered gravity on different cell types and organisms. Real-time imaging during simulations are of particular interest as we can study how basic cell functions such as cell division, cell migration, and proliferation progress under microgravity conditions. However, design limitations of present microgravity simulators such as susceptibility to parasitic vibration and displacement of the sample from the center of rotation challenge the accuracy of experiment results and live images. We have developed a cell culture sample holder module suitable for live microscopic imaging on an RPM. CAD modeling and 3D printing technology were used to implement modifications to the sample holder and to install a digital microscope to perform live bright-field and fluorescent imaging. Vibration damping materials were also investigated to allow for stable imaging while the microgravity simulator was within a cell culture incubator. Novel methods and hardware modifications for improving live cell imaging on ground-based microgravity simulators were proposed and discussed.

3D printing↗

Allochthonous Addition of Meteoritic Organics to the Lunar Regolith

Preparation of lunar samples 74220,861 was discussed in detail in [3, 4]. Our analysis sequence was as follows: optical microscopy, UV fluorescence imaging, -Raman, FESEM-EDX imaging and mapping, FETEMEDX imaging and mapping of a Focused Ion Beam (FIB) extracted section, and NanoSIMs analysis. We observed fluffytextured C-rich regions of interest (ROI) on three different volcanic glass beads. Each ROI was several m2 in size and fluoresced when exposed to UV. Using FESEM/EDX, the largest ROI measured ~36 m and was located on an edge of a plateau located on the uppermost surface of the bead. The ROI was covered on one edge by a siliceous filament emanating from the plateau surface indicating it was attached to the bead while on the Moon. EDX mapping of the ROI shows it is composed primarily of heterogeneously distributed C. Embedded with the carbonaceous phase are localized concentrations of Si, Fe, Al and Ti indicating the presence of glass and/or minerals grains. -Raman showed strong D- and G-bands and their associated second order bands; intensity and location of these bands indicates the carbonaceous matter is structurally disorganized. A TEM thin section was extracted from the surface of a glass bead using FIB microscopy. High resolution TEM imaging and selected area electron diffraction demonstrate the carbonaceous layer to be amorphous; it lacked any long or short range order characteristic of micro- or nanocrystalline graphite. Additionally TEM imaging also revealed the presence of submicron mineral grains, typically < 50 nm in size, dispersed within the carbonaceous layer. NanoSIMs data will be presented and discussed at the meeting. Given the noted similarities between the carbonaceous matter present on 74220 glass beads and meteoritic kerogen, we suggest the allochthonous addition of meteoritic organics as the most probable source for the C-rich ROIs.

Thomas-Keprta, K. L.↗

Rapid in situ hybridization technique using 16S rRNA segments for detecting and differentiating the closely related gram-positive organisms Bacillus polymyxa and Bacillus macerans

A rapid, sensitive, inexpensive in situ hybridization technique, using 30-mer 16S rRNA probes, can specifically differentiate two closely related Bacillus spp., B. polymyxa and B. macerans. The 16S rRNA probes were labeled with a rhodamine derivative (Texas Red), and quantitative fluorescence measurements were made on individual bacterial cells. The microscopic fields analyzed were selected by phase-contrast microscopy, and the fluorescence imaging analyses were performed on 16 to 67 individual cells. The labeled 16S rRNA probe, POL, whose sequence was a 100% match with B. polymyxa 16S rRNA but only a 60% match with B. macerans 16S rRNA, gave quantitative fluorescence ratio measurements that were 34.8-fold higher for B. polymyxa cells than for B. macerans cells. Conversely, the labeled probe, MAC, which matched B. polymyxa 16S rRNA in 86.6% of its positions and B. macerans 16S rRNA in 100% of its positions, gave quantitative fluorescence measurements that were 59.3-fold higher in B. macerans cells than in B. polymyxa cells. Control probes, whose 16S rRNA sequence segment (P-M) was present in both B. polymyxa and B. macerans as well as a panprokaryotic probe (16S), having a 100% match with all known bacteria, hybridized equally well with both organisms. These latter hybridizations generated very high fluorescence signals, but their comparative fluorescence ratios (the differences between two organisms) were low. The control paneukaryotic probe (28S), which had less than 30% identity for both B. macerans and B. polymyxa, did not hybridize with either organism.

Non-NASA Center↗

Selective functionalization of carbon nanotube tips allowing fabrication of new classes of nanoscale sensing and manipulation tools

Embodiments in accordance with the present invention relate to techniques for the growth and attachment of single wall carbon nanotubes (SWNT), facilitating their use as robust and well-characterized tools for AFM imaging and other applications. In accordance with one embodiment, SWNTs attached to an AFM tip can function as a structural scaffold for nanoscale device fabrication on a scanning probe. Such a probe can trigger, with nanometer precision, specific biochemical reactions or conformational changes in biological systems. The consequences of such triggering can be observed in real time by single-molecule fluorescence, electrical, and/or AFM sensing. Specific embodiments in accordance with the present invention utilize sensing and manipulation of individual molecules with carbon nanotubes, coupled with single-molecule fluorescence imaging, to allow observation of spectroscopic signals in response to mechanically induced molecular changes. Biological macromolecules such as proteins or DNA can be attached to nanotubes to create highly specific single-molecule probes for investigations of intermolecular dynamics, for assembling hybrid biological and nanoscale materials, or for developing molecular electronics. In one example, electrical wiring of single redox enzymes to carbon nanotube scanning probes allows observation and electrochemical control over single enzymatic reactions by monitoring fluorescence from a redox-active cofactor or the formation of fluorescent products. Enzymes ''nanowired'' to the tips of carbon nanotubes in accordance with embodiments of the present invention, may enable extremely sensitive probing of biological stimulus-response with high spatial resolution, including product-induced signal transduction.

Wade, Lawrence A.↗

Foliar Reflectance and Fluorescence Responses for Plants Under Nitrogen Stress Determined with Active and Passive Systems

Vegetation productivity is driven by nitrogen (N) availability in soils. Both excessive and low soil N induce physiological changes in plant foliage. In 2001, we examined the use of spectral fluorescence and reflectance measurements to discriminate among plants provided different N fertilizer application rates: 20%, 50%, 100% and 150% of optimal N levels. A suite of optical, fluorescence, and biophysical measurements were collected on leaves from field grown corn (Zea mays L.) and soybean plants (Glycine max L.) grown in pots (greenhouse + ambient sunlight daily). Three types of steady state laser-induced fluorescence measurements were made on adaxial and abaxial surfaces: 1) fluorescence images in four 10 nm bands (blue, green, red, far-red) resulting from broad irradiance excitation; 2) emission spectra (5 nm resolution) produced by excitation at single wavelengths (280,380 or 360, and 532 nm); and 3) excitation spectra (2 nm resolution), with emission wavelengths fixed at wavelengths centered on selected solar Fraunhofer lines (532,607,677 and 745 nm). Two complementary sets of high resolution (less than 2 nm) optical spectra were acquired for both adaxial and abaxial leaf surfaces: 1) optical properties (350-2500 nm) for reflectance, transmittance, and absorptance; and 2) reflectance spectra (500-1000 nm) acquired with and without a short pass filter at 665 nm to determine the fluorescence contribution to apparent reflectance in the 650-750 spectrum, especially at the 685 and 740 nm chlorophyll fluorescence (ChIF) peaks. The strongest relationships between foliar chemistry and optical properties were demonstrated for C/N content and two optical parameters associated with the red edge inflection point. Select optical properties and ChIF parameters were highly correlated for both species. A significant contribution of ChIF to apparent reflectance was observed, averaging 10-25% at 685 nm and 2 - 6% at 740 nm over all N treatments. Discrimination of N treatment groups was possible with specific fluorescence band ratios (e.g., F740/F525 obtained with 380EX). From all measurements assessing fluorescence, higher ChIF and blue/green emissions were measured from the abaxial leaf surfaces; Abaxial surfaces also produced higher reflectances in the 400-800 nm spectrum. Fluorescence information collected in Fraunhofer regions located on the shoulders of ChIF features compared favorably with peak emissions. This supports the potential capability of a future space-born interferometer sensor to capture plant canopy fluorescence.

Middleton, E. M.↗

Flow-Tagging Velocimetry for Hypersonic Flows Using Fluorescence of Nitric Oxide

We demonstrate a new variation of molecular-tagging velocimetry for hypersonic flows based on laser-induced fluorescence. A thin line of nitric-oxide molecules is excited with a laser beam and then, after a time delay, a fluorescence image of the displaced line is acquired. One component of velocity is determined from the time of flight. This method is applied to measure the velocity profile in a Mach 8.5 laminar, hypersonic boundary layer in the Australian National University s T2 free-piston shock tunnel. The single-shot velocity measurement uncertainty in the freestream was found to be 3.5%, based on 90% confidence. The method is also demonstrated in the separated flow region forward of a blunt fin attached to a flat plate in a Mach 7.4 flow produced by the Australian National University s T3 free-piston shock tunnel. The measurement uncertainty in the blunt fin experiment is approximately 30%, owing mainly to low fluorescence intensities, which could be improved significantly in future experiments. This velocimetry method is applicable to very high-speed flows that have low collisional quenching of the fluorescing species. It is particularly convenient in facilities where planar laser-induced fluorescence is already being performed.

Danehy, Paul M.↗

Alternative Bio-Derived JP-8 Class Fuel and JP-8 Fuel: Flame Tube Combustor Test Results Compared using a GE TAPS Injector Configuration

This paper presents results from tests in a flame tube facility, where a bio-derived alternate fuel was compared with JP-8 for emissions and general combustion performance. A research version of General Electric Aviation (GE) TAPS injector was used for the tests. Results include combustion efficiency from gaseous emission measurements, 2D planar laser-based imaging as well as basic flow visualization of the flame. Four inlet test conditions were selected that simulate various engine power conditions relevant to NASA Fundamental Aeronautics Supersonics Project and Environmentally Responsible Aviation Program. One inlet condition was a pilot-only test point. The other three inlet conditions incorporated fuel staging via a split between the pilot and main circuits of either 10%/90% or 20%/80%. For each engine power condition, three fuel mixes were used: 100% JP-8; 100% alternative; and a blend of the two, containing 75% alternative. Results for the inlet cases that have fuel split between pilot and main, indicate that fuel from the pilot appears to be evaporated by the time it reaches the dome exit. Main circuit liquid evaporates within a downstream distance equal to annulus height, no matter the fuel. Some fuel fluorescence images for a 10%/90% fuel staging case show a distinct difference between JP-8 and bio-derived fuel. OH PLIF results indicate that OH forms in a region more centrally-located for the JP-8 case downstream of the pilot, in its central recirculation region (CRZ). For the bio-derived Hydrotreated Renewable Jet (HRJ) fuel, however, we do not see much OH in the CRZ. The OH image structure near the dome exit is similar for the two fuels, but farther downstream the OH in the CRZ is much more apparent for the JP-8 than for the alternate fuel. For all conditions, there was no discernable difference between fuel types in combustion efficiency or emissions.

gas turbine combustors↗

Velocity profiles in a hot jet by simplified RELIEF

Raman Excitation + Laser Induced Electronic Fluorescence (RELIEF) is a double resonance velocimetry technique in which oxygen molecules are vibrationally excited via stimulated Raman scattering at a specific location within a flow field. After suitable time delay, typically 1-10 microseconds, the displacement of the tagged molecules is determined by laser induced fluorescence imaging. Providing support for the installation of simplified RELIEF flow tagging instrumentation at NASA LaRC was the principal goal of this research.

Source record↗

Gamma Ray and Neutron Spectrometer for the Lunar Resource Mapper

One of the early Space Exploration Initiatives will be a lunar orbiter to map the elemental composition of the Moon. This mission will support further lunar exploration and habitation and will provide a valuable dataset for understanding lunar geological processes. The proposed payload will consist of the gamma ray and neutron spectrometers which are discussed, an x ray fluorescence imager, and possibly one or two other instruments.

Moss, C. E.↗

Nonintrusive, multipoint velocity measurements in high-pressure combustion flows

A combined experimental and analytical effort was conducted to demonstrate the applicability of OH Doppler-shifted fluorescence imaging of velocity distributions in supersonic combustion gases. The experiments were conducted in the underexpanded exhaust flow from a 6.8 atm, 2400 K, H2-O2-N2 burner exhausting into the atmosphere. In order to quantify the effects of in-plane variations of the gas thermodynamic properties on the measurement accuracy, a set of detailed measurements of the OH (1,0) band collisional broadening and shifting in H2-air gases was produced. The effect of pulse-to-pulse variations in the dye laser bandshape was also examined in detail and a modification was developed which increased in the single pulse bandwidth, thereby increasing the intraimage velocity dynamic range as well as reducing the sensitivity of the velocity measurement to the gas property variations. Single point and imaging measurements of the velocity field in the exhaust flowfield were compared with 2D, finite-rate kinetics simulations of the flowfield. Relative velocity accuracies of +/- 50 m/s out of 1600 m/s were achieved in time-averaged imaging measurements of the flow over an order of magnitude variation in pressure and a factor of two variation in temperature.

Allen, M.↗

Experimental Reacting Hydrogen Shear Layer Data at High Subsonic Mach Number

The flow in a planar shear layer of hydrogen reacting with hot air was measured with a two-component laser Doppler velocimeter (LDV) system, a schlieren system, and OH fluorescence imaging. It was compared with a similar air-to-air case without combustion. The high-speed stream's flow speed was about 390 m/s, or Mach 0.71, and the flow speed ratio was 0.34. The results showed that a shear layer with reaction grows faster than one without; both cases are within the range of data scatter presented by the established data base. The coupling between the streamwise and the cross-stream turbulence components inside the shear layers was low, and reaction only increased it slightly. However, the shear layer shifted laterally into the lower speed fuel stream, and a more organized pattern of Reynolds stress was present in the reaction shear layer, likely as a result of the formation of a larger scale structure associated with shear layer corrugation from heat release. Dynamic pressure measurements suggest that coherent flow perturbations existed inside the shear layer and that this flow became more chaotic as the flow advected downstream. Velocity and thermal variable values are listed in this report for a computational fluid dynamics (CFD) benchmark.

Chang, C. T.↗

Effects of Passive Fuel-Air Mixing Control on Burner Emissions Via Lobed Fuel Injectors

The present experimental study examines the effects of differing levels of passive fuel-air premixing on flame structures and their associated NO(x) and CO emissions. Four alternative fuel injector geometries were explored, three of which have lobed shapes. These lobed injectors mix fuel and air and strain species inter-faces to differing extents due to streamwise vorticity generation, thus creating different local or core equivalence ratios within flow regions upstream of flame ignition and stabilization. Prior experimental studies of two of these lobed injector flowfields focused on non-reactive mixing characteristics and emissions measurements for the case where air speeds were matched above and below the fuel injector, effectively generating stronger streamwise vorticity than spanwise vorticity. The present studies examine the effects of airstream mismatch (and hence additional spanwise vorticity generation), effects of confinement of the crossflow to reduce the local equivalence ratio, and the effects of altering the geometry and position of the flameholders. NO(x) and CO emissions as well as planar laser-induced fluorescence imaging (PLIF) of seeded acetone are used to characterize injector performance and reactive flow evolution.

Mitchell, M. G.↗

Quantitative Measurements of Nitric Oxide Concentration in High-Pressure, Swirl-Stabilized Spray Flames

Lean direct-injection (LDI) spray flames offer the possibility of reducing NO(sub x) emissions from gas turbines by rapid mixing of the liquid fuel and air so as to drive the flame structure toward partially-premixed conditions. We consider the technical approaches required to utilize laser-induced fluorescence methods for quantitatively measuring NO concentrations in high-pressure LDI spray flames. In the progression from atmospheric to high-pressure measurements, the LIF method requires a shift from the saturated to the linear regime of fluorescence measurements. As such, we discuss quantitative, spatially resolved laser-saturated fluorescence (LSF), linear laser-induced fluorescence (LIF), and planar laser-induced fluorescence (PLIF) measurements of NO concentration in LDI spray flames. Spatially-resolved LIF measurements of NO concentration (ppm) are reported for preheated, LDI spray flames at pressures of two to five atmospheres. The spray is produced by a hollow-cone, pressure-atomized nozzle supplied with liquid heptane. NO is excited via the Q(sub 2)(26.5) transition of the gamma(0,0) band. Detection is performed in a two nanometer region centered on the gamma(0,1) band. A complete scheme is developed by which quantitative NO concentrations in high-pressure LDI spray flames can be measured by applying linear LIF. NO is doped into the reactants and convected through the flame with no apparent destruction, thus allowing a NO fluorescence calibration to be taken inside the flame environment. The in-situ calibration scheme is validated by comparisons to a reference flame. Quantitative NO profiles are presented and analyzed so as to better understand the operation of lean-direct injectors for gas turbine combustors. Moreover, parametric studies are provided for variations in pressure, air-preheat temperature, and equivalence ratio. Similar parametric studies are performed for lean, premixed-prevaporized flames to permit comparisons to those for LDI flames. Finally, PLIF is expanded to high pressure in an effort to quantify the detected fluorescence image for LDI flames. Success is achieved by correcting the PLIF calibration via a single-point LIF measurement. This procedure removes the influence of any preferential background that occurs in the PLIF detection window. In general, both the LIF and PLIF measurements verify that the LDI strategy could be used to reduce NO(sub x) emissions in future gas turbine combustors.

Cooper, Clayton S.↗

Microscale Concentration Measurements Using Laser Light Scattering Methods

The development of lab-on-a-chip devices for microscale biochemical assays has led to the need for microscale concentration measurements of specific analyses. While fluorescence methods are the current choice, this method requires developing fluorophore-tagged conjugates for each analyte of interest. In addition, fluorescent imaging is also a volume-based method, and can be limiting as smaller detection regions are required.

Niederhaus, Charles↗

The Focal Surface of the JEM-EUSO Telescope

Extreme Universe Space Observatory onboard JEM/EP (JEM-EUSO) is a space mission to study extremely high-energy cosmic rays. The JEM-EUSO instrument is a wide-angle refractive telescope in near-ultraviolet wavelength region to observe time-resolved atmospheric fluorescence images of the extensive air showers from the International Space Station. The focal surface is a spherical curved surface, and its area amounts to about 4.5 square m. The focal surface detector is covered with about 6,000 multi-anode photomultipliers (MAPMTs). The focal surface detector consists of Photo-Detector-Modules, each of which consists of 9 Elementary Cells (ECs). The EC contains 4 units of the MAPMTs. Therefore, about 1,500 ECs or about 160 PDMS are arranged on the whole of the focal surface of JEM- EUSO. The EC is a basic unit of the front-end electronics. The PDM is a, basic unit of the data acquisition system

Kawasaki, Yoshiya↗

Advancement of the Wide-angle JEM-EUSO Optical System with Holographic and Fresnel Lenses

JEM-EUSO is a space mission to observe extremely high-energy cosmic rays, evolved from the previous design studies of EUSO. It is adjusted for the Japan Experiment Module (JEM) of the International Space Station (ISS). JEM-EUSO uses a wide-angle refractive telescope in near-ultraviolet wavelength region to observe from ISS the time-and-space-resolved atmospheric fluorescence images of the extensive air showers. The JEM-EUSO optics is re-designed after the ESA-Phase A studies to upgrade the light-collecting-power by using a new material CYTOP, and its overall light-collecting power is about 1.5 times higher than the ESA-Phase A baseline optics. We describe in this paper an optimized optics design that maximizes the sensitivity of JEM-EUSO, and the results of the optics manufacturing tests.

Takizawa, Y.↗