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At least 73 records · Page 4

Miniature Sensor Probe for O2, CO2, and H2O Monitoring in Portable Life Support Systems

A miniature sensor probe, composed of four sensors which monitor the partial pressure of O2, CO2, H2O, and temperature, designed to operate in the portable life support system (PLSS), has been demonstrated. The probe provides an important advantage over existing technology in that it is able to operate reliably while wet. These luminescence-based fiber optic sensors consist of an indicator chemistry immobilized in a polymeric film, whose emission lifetime undergoes a strong change upon a reversible interaction with the target gas. Each sensor includes chemistry specifically sensitive to one target parameter. All four sensors are based on indicator chemistries that include luminescent dyes from the same chemical family, and therefore exhibit similar photochemical properties, which allow performing measurements of all the sensors by a single, compact, low-power optoelectronic unit remotely connected to the sensors by an electromagnetic interference-proof optical fiber cable. For space systems, using these miniature sensor elements with remote optoelectronics provides unmatched design flexibility for measurements in highly constrained volume systems such as the PLSS. A 10 mm diameter and 15 mm length prototype multiparameter probe was designed, fabricated, tested, and demonstrated over a wide operational range of gas concentration, humidity, and temperature relevant to operation in the PLSS. The sensors were evaluated for measurement range, precision, accuracy, and response time in temperatures ranging from 50 aF-150 aF and relative humidity from dry to 100% RH. Operation of the sensors in water condensation conditions was demonstrated wherein the sensors not only tolerated liquid water but actually operated while wet.

Delgado, Jesus

Automated Lab-on-a-Chip Electrophoresis System

Capillary electrophoresis is an analytical technique that can be used to detect and quantify extremely small amounts of various biological molecules. In the search for biochemical traces of life on other planets, part of this search involves an examination of amino acids, which are the building blocks of life on Earth. The most sensitive method for detecting amino acids is the use of laser induced fluorescence. However, since amino acids do not, in general, fluoresce, they first must be reacted with a fluorescent dye label prior to analysis. After this process is completed, the liquid sample then must be transported into the electrophoresis system. If the system is to be reused multiple times, samples must be added and removed each time. In typical laboratories, this process is performed manually by skilled human operators using standard laboratory equipment. This level of human intervention is not possible if this technology is to be implemented on extraterrestrial targets. Microchip capillary electrophoresis (CE) combined with laser induced fluorescence detection (LIF) was selected as an extremely sensitive method to detect amino acids and other compounds that can be tagged with a fluorescent dye. It is highly desirable to package this technology into an integrated, autonomous, in situ instrument capable of performing CE-LIF on the surface of an extraterrestrial body. However, to be fully autonomous, the CE device must be able to perform a large number of sample preparation and analysis operations without the direct intervention of a human.

Willis, Peter A.

OH Planar Laser-Induced Fluorescence from Microgravity Droplet Combustion

Droplet combustion under microgravity conditions has been extensively studied, but laser diagnostics have just begun to be employed in microgravity droplet experiments. This is due in part to the level of difficulty associated with laser system size, power and economic availability. Hydroxyl radical (OH) is an important product of combustion, and laser-induced fluorescence (LIF) has proved to be an adequate and sensitive tool to measure OH. In this study, a frequency doubled Nd:YAG laser and a doubled dye laser, compact and reliable enough to perform OH PLIF experiments aboard a parabolic flight-path aircraft, has been developed and successfully demonstrated in a methanol droplet flame experiment. Application to microgravity conditions is planned aboard parabolic flight-path aircraft.

Winter, Michael

Micromotional studies of utricular and canal afferents

The long-range goal of this research was to refine our understanding of the sensitivity of the vestibular components of the ear to very-low-amplitude motion, especially, the role of gravity in this sensitivity. We focused on the American bullfrog--a common animal subject for vestibular sensory research. Our principal experimental method was to apply precise, sinusoidal microrotational stimuli to an anesthetized animal subject, to record the resulting responses in an individual vestibular nerve fiber from the intact ear, and to use intracellular dye to trace the fiber and thus identify the vestibular sensor that gave rise to it. In this way, we were able to identify specific micromotional sensitivities and to associate those sensitivities definitely with specific sensors. Furthermore, by recording from nerve fibers after they leave the intact inner-ear cavity, we were able to achieve these identifications without interrupting the delicate micromechanics of the inner ear. We were especially concerned with the relative roles of the utricle and the anterior semicircular canal in the sensing of microrotational motion of the head about horizontal axes, and with the role of gravity in mediating that sensing process in the utricle. The functional characterization of individual nerve fibers was accomplished with a conventional analytical tool, the cycle histogram, in which the nerve impulse rate was plotted against the phase of the sinusoidal stimulus.

Lewis, Edwin R.

Mechanical continuity and reversible chromosome disassembly within intact genomes removed from living cells

Chromatin is thought to be structurally discontinuous because it is packaged into morphologically distinct chromosomes that appear physically isolated from one another in metaphase preparations used for cytogenetic studies. However, analysis of chromosome positioning and movement suggest that different chromosomes often behave as if they were physically connected in interphase as well as mitosis. To address this paradox directly, we used a microsurgical technique to physically remove nucleoplasm or chromosomes from living cells under isotonic conditions. Using this approach, we found that pulling a single nucleolus or chromosome out from interphase or mitotic cells resulted in sequential removal of the remaining nucleoli and chromosomes, interconnected by a continuous elastic thread. Enzymatic treatments of interphase nucleoplasm and chromosome chains held under tension revealed that mechanical continuity within the chromatin was mediated by elements sensitive to DNase or micrococcal nuclease, but not RNases, formamide at high temperature, or proteases. In contrast, mechanical coupling between mitotic chromosomes and the surrounding cytoplasm appeared to be mediated by gelsolin-sensitive microfilaments. Furthermore, when ion concentrations were raised and lowered, both the chromosomes and the interconnecting strands underwent multiple rounds of decondensation and recondensation. As a result of these dynamic structural alterations, the mitotic chains also became sensitive to disruption by restriction enzymes. Ion-induced chromosome decondensation could be blocked by treatment with DNA binding dyes, agents that reduce protein disulfide linkages within nuclear matrix, or an antibody directed against histones. Fully decondensed chromatin strands also could be induced to recondense into chromosomes with pre-existing size, shape, number, and position by adding anti-histone antibodies. Conversely, removal of histones by proteolysis or heparin treatment produced chromosome decondensation which could be reversed by addition of histone H1, but not histones H2b or H3. These data suggest that DNA, its associated protein scaffolds, and surrounding cytoskeletal networks function as a structurally-unified system. Mechanical coupling within the nucleoplasm may coordinate dynamic alterations in chromatin structure, guide chromosome movement, and ensure fidelity of mitosis.

NASA Discipline Cell Biology

BioSentinel: An Adaptable Platform for Studying the Biological Effects of Deep Space Radiation

NASA's BioSentinel mission is a 6U nanosatellite with autonomous life support that will utilize the budding yeast Saccharomyces cerevisiae to study the DNA damage response to the deep space radiation environment. BioSentinel is planned to launch in 2019 as a secondary payload on the Space Launch System's first Exploration Mission (EM-1), and will undergo a lunar fly-by and enter heliocentric orbit after deployment. As the first biological mission beyond Low Earth Orbit (LEO) in nearly half a century, this mission will help fill critical gaps in knowledge about the effects of uniquely composed, chronic, low-flux deep space radiation on biological systems. Yeast is well-suited for this mission due to its desiccation tolerance and space-flight heritage. As a eukaryotic model organism, it also serves as a robust analog for human cells. Data gathered on this mission will thus inform us of the hazards involved in long-duration human exploration in deep space, and the protections necessary to mitigate them. Due to its low-cost, flexible and advanced technology, the 4U BioSensor payload contained within the nanosatellite is adaptable to other model microorganisms, exploration platforms and environments relevant to human exploration, such as the ISS, the Lunar Orbital Platform - Gateway and future lunar landers. In order to query the DNA damage response to deep space radiation, BioSentinel contains a wild type yeast strain as a positive control, and a radiation sensitive rad51 mutant strain that is defective for DNA repair. Yeast cells are desiccated in microfluidic cards, and rehydrated with growth medium and metabolic indicator dye at the desired time points during the mission. A thermal control system supports these stasis and growth states, and an optical system continuously measures cell growth and metabolism. An onboard radiation spectrometer and dosimeter allows us to correlate the dose, energy and particle-type of deep space radiation to the biological response. Data received from the deep space biosensor will be compared to control payloads on Earth and the ISS. Ongoing science testing for the BioSentinel project includes optimization for cell viability, desiccation tolerance, and long-term biocompatibility, as well as radiation experiments to understand the sensitivity and responsiveness of cells to varying radiation doses and particle types.

BioSentinel

Three color laser fluorometer for studies of phytoplankton fluorescence

A three-color laser fluorometer has been developed for field work operations. Using two tunable dye lasers (excitation wavelengths at 440 nm and 530 nm), broadband wavelength optical filters were selected to obtain maximum fluorescence sensitivity at wavelengths greater than 675 nm (chlorophyll) and 575 + or - 15 nm (phycoerythrin). The laser fluorometer permits the measurement of phytoplankton pigments under static or flowing conditions and more closely resembles the time scales (ns) and energy levels (mW) of other laser-induced fluorescence instruments.

Phinney, David A.

Single-Arm Double-Mode Double-Order Planar Waveguide Interferometric Sensor

We have met the goals stated in section one for the project. We have demonstrated the feasibility of a single-arm double-mode double-order waveguide interferometer as a cost efficient alternative to an optical chemical sensor. Experimental prototype was built as a dye-doped polymer waveguide with propagating modes of orders <<0>> and <<1>> of the same TM polarization. The prototype demonstrated sensitivity to ammonia of the order of 200 ppm per one full oscillation of the signal. Sensor based on polyimide doped with BCP can operate at elevated temperature up to 150 C. Upon the future funding, we are planning to optimize the light source, material and the design in order to achieve sensitivity of the order of 1 ppm per full oscillations.

Sarkisov, Sergey S.

Portable Unit for Metabolic Analysis

The Portable Unit for Metabolic Analysis (PUMA) is an instrument that measures several quantities indicative of human metabolic function. Specifically, this instrument makes time-resolved measurements of temperature, pressure, flow, and the partial pressures of oxygen and carbon dioxide in breath during both inhalation and exhalation. Portable instruments for measuring these quantities have been commercially available, but the response times of those instruments are too long to enable temporal resolution of phenomena on the time scales of human respiration cycles. In contrast, the response time of the PUMA is significantly shorter than characteristic times of human respiration phenomena, making it possible to analyze varying metabolic parameters, not only on sequential breath cycles but also at successive phases of inhalation and exhalation within the same breath cycle. In operation, the PUMA is positioned to sample breath near the subject s mouth. Commercial off-the-shelf sensors are used for three of the measurements: a miniature pressure transducer for pressure, a thermistor for temperature, and an ultrasonic sensor for flow. Sensors developed at Glenn Research Center are used for measuring the partial pressures of oxygen and carbon dioxide: The carbon dioxide sensor exploits the relatively strong absorption of infrared light by carbon dioxide. Light from an infrared source passes through the stream of inhaled or exhaled gas and is focused on an infrared- sensitive photodetector. The oxygen sensor exploits the effect of oxygen in quenching the fluorescence of ruthenium-doped organic molecules in a dye on the tip of an optical fiber. A blue laser diode is used to excite the fluorescence, and the optical fiber carries the fluorescent light to a photodiode, the temporal variation of the output of which bears a known relationship with the rate of quenching of fluorescence and, hence, with the partial pressure of oxygen. The outputs of the sensors are digitized, preprocessed by a small onboard computer, and then sent wirelessly to a desktop computer, where the collected data are analyzed and displayed. In addition to the raw data on temperature, pressure, flow, and mole fractions of oxygen and carbon dioxide, the display can include volumetric oxygen consumption, volumetric carbon dioxide production, respiratory equivalent ratio, and volumetric flow rate of exhaled gas.

Dietrich, Daniel L.

Hair cell tufts and afferent innervation of the bullfrog crista ampullaris

Within the bullfrog semicircular canal crista, hair cell tuft types were defined and mapped with the aid of scanning electron microscopy. Dye-filled planar afferent axons had mean distal axonal diameters of 1.6-4.9 microns, highly branched arbors, and contacted 11-24 hair cells. Dye-filled isthmus afferent axons had mean distal axonal diameters of 1.8-7.9 microns, with either small or large field arbors contacting 4-9 or 25-31 hair cells. The estimated mean number of contacts per innervated hair cell was 2.2 for planar and 1.3 for isthmus afferent neurons. Data on evoked afferent responses were available only for isthmus units that were observed to respond to our microrotational stimuli. Of 21 such afferent neurons, eight were successfully dye-filled. Within this sample, high-gain units had large field arbors and lower-gain units had small field arbors. The sensitivity of each afferent neuron was analyzed in terms of noise equivalent input (NEI), the stimulus amplitude for which the afferent response amplitude is just equivalent to the rms deviation of the instantaneous spike rate. NEI for isthmus units varied from 0.63 to 8.2 deg/s; the mean was 3.2 deg/s.

Myers, Steven F.

Tea ( Camellia sinensis ) Extract-Mediated Green Synthesis of Co 3 O 4 and Co 3 O 4 @Graphene Nanocomposites for Multifunctional Applications in Pollutant Degradation, Sensing, and Energy Storage

A novel solution-mixing method was proposed to synthesize Co 3 O 4 /graphene nanocomposites (Co 3 O 4 @Gr) using a green tea leaf (Camellia sinensis) extract as the reducing agent. XRD analysis shows that the as-prepared Co 3 O 4 @Gr exhibits a cubic spinel crystal structure. From morphological analysis, the obtained Co 3 O 4 NS forms spherical clusters that are uniformly distributed on the graphene surface. FT-IR and Raman analyses confirmed the strong molecular and vibrational interactions between the Co 3 O 4 NS and Gr. The suppressed PL intensity peak of the Co 3 O 4 @Gr NCs indicated significant inhibition in the recombination of charge carriers between the hybrid orbitals within the composites. As a result, the catalytic efficiency of Co 3 O 4 @Gr NCs increased to 80% compared to pristine Co 3 O 4 , which exhibited only 45% efficiency against methylene blue (MB) dye. Moreover, the as-prepared NCs exhibited a detection limit of 0.01−224 μM, demonstrating a superior low-DPA detection with high sensitivity. The Co 3 O 4 @Gr/GCE exhibits admirable selectivity for various pesticides, fungicides, and metal ions, with outstanding reproducibility and stability. From electrochemical investigations, the highest specific capacitance values of the as-synthesized Co 3 O 4 @Gr were 349 F/g at a scan rate of 5 mV/s and 158 F/g at a current density of 1 A/g.

Capacitors

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel

The effect of low temperature on poly(3-methyl- N -vinylcaprolactam)- b -poly( N -vinylpyrrolidone) diblock copolymer nanovesicles assembled from all-aqueous media

Nanosized polymeric vesicles (polymersomes) self-assembled from double hydrophilic copolymers of poly(3-methyl-N-vinylcaprolactam) n -b-poly(N-vinylpyrrolidone) m (PMVC n -b-PVPON m ) using all aqueous media are a promising platform for biomedical applications, because of their superior stability over liposomes in vivo and high loading capacity. Herein, we explored the temperature-sensitive behavior of PMVC 58 -b-PVPON 65 vesicles using transmission electron microscopy (TEM), dynamic light scattering (DLS), atomic force microscopy (AFM), and small-angle neutron scattering (SANS) in response to lowering the solution temperature from 37 to 25, 20, 14 and 4 °C. The copolymer vesicles with an average size of 350 nm at 37 °C were assembled from the diblock copolymer dissolved in aqueous solution at 4 °C. We show that while the polymersome's size gradually decreases upon the temperature decrease from 37 to 4 °C, the average shell thickness increases from 17 nm to 25 nm, respectively. SANS study revealed that the PMVC 58 -b-PVPON 65 vesicle undergoes a gradual structure evolution from a dense-shell vesicle at 37–25 °C to a highly-hydrated shell vesicle at 20–14 °C to molecular chain aggregates at 4 °C. From SANS contrast matching study, this vesicle behavior is found to be driven by the gradual rehydration of PMVC block at 37–14 °C. The shell hydration at 20–14 °C also correlated with the 4.4-fold decrease in the relative fluorescence intensity from vesicle-encapsulated fluorescent dye, indicating ~80% of the dye release within 12 hours after the vesicle exposure to 14 °C. No significant (<5%) dye release was observed for the vesicle solutions at 37–20 °C, indicating excellent cargo retention inside the vesicles. Our study provides new fundamental insights on temperature-sensitive polymer vesicles and demonstrates that the copolymer assembly into polymersomes can be achieved by decreasing a copolymer aqueous solution temperature below 14 °C followed by solution exposure to ≥20 °C. This type of all-aqueous assembly, instead of nanoprecipitation from organic solvents or solvent exchange, can be highly desirable for encapsulating a wide range of biological molecules, including proteins, peptides, and nucleic acids, into stable polymer vesicles without a need for organic solvents for dissolution of the copolymers that are amphiphilic at physiologically relevant temperatures of 20–37 °C.

36 MATERIALS SCIENCE

Development of Thermally Stable and Highly Fluorescent IR Dyes

Fluorophores are the core component in various optical applications such as sensors and probes. Fluorphores with low-energy or long wavelength emission, in particular, in NIR region, possess advantages of low interference and high sensitivity. In this study, we has explored several classes of imidazole-based compounds for NIR fluorescent properties and concluded: (1) thiazole-based imidazole compounds are fluorescent; (2) emission energy is tunable by additional donor groups; (3) they also possess impressive two- photon absorption properties; and (4) fluorescence emission can be induced by two- photon input. This report summarizes (1) synthesis of new series of fluorophore; (2) impact of electron-withdrawing groups on fluorescent property; (3) unique property of two-photon absorption; and (4) on-going development.

Bu, Xiu R.

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

SO2 Spectroscopy with A Tunable UV Laser

A portion of the fluorescence spectrum of SO2 has been studied using a narrow wavelength doubled dye laser as the exciting source. One purpose of this study is to evaluate the use of SO2 resonance re-emission as a probe of SO2 in the atmosphere. When the SO2 is excited by light at 300.2 nm, for example, a strong reemission peak is observed which is Stokes-shifted from the incident light wavelength by the usual Raman shift (the VI symmetric vibration frequency 1150.5/cm ). The intensity of this peak is sensitive to small changes (.01 nm) in the incident wavelength. Measurements of the N2 quenching and self quenching of this re-emission have been obtained. Preliminary analysis of this data indicates that the quenching is weak but not negligible. The dye laser in our system is pumped by a pulsed N2 laser. Tuning 'and spectral narrowing are accomplished using a telescope-echelle grating combination. In a high power configuration the resulting pulses have a spectral width of about 5 x 10(exp -3) nm and a time duration of about 6 nsec. The echelle grating is rotated by a digital stepping motor, such that each step shifts the wavelength by 6 x 10(exp -4) nm. In addition to the tunable, narrow wavelength uv source and spectral analysis of the consequent re-emission, the system also provides time resolution of the re-emitted light to 6 nsec resolution. This capability is being used to study the lifetime of low pressure S02 fluorescence at different wavelengths and pressures.

Morey, W. W.