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At least 37 records · Page 2

Lunar Dust and Lunar Simulant Activation and Monitoring

Prior to returning to the moon, understanding the effects of lunar dust on both human physiology and mechanical equipment is a pressing concern, as problems related to lunar dust during the Apollo missions have been well documented (J.R. Gaier, The Effects of Lunar Dust on EVA Systems During the Apollo Missions. 2005, NASA-Glenn Research Center. p. 65). While efforts were made to remove the dust before reentering the lunar module, via brushing of the suits or vacuuming, a significant amount of dust was returned to the spacecraft, causing various problems. For instance, astronaut Harrison Schmitt complained of hay fever effects caused by the dust, and the abrasive nature of the material was found to cause problems with various joints and seals of the spacecraft and suits. It is clear that, in order to avoid potential health and performance problems while on the lunar surface, the reactive properties of lunar dust must be quenched. It is likely that soil on the lunar surface is in an activated form, i.e. capable of producing oxygen-based radicals in a humidified air environment, due to constant exposure to meteorite impacts, UV radiation, and elements of the solar wind. An activated silica surface serves as a good example. An oxygen-based radical species arises from the breaking of Si-OSi bonds. This system is comparable to that expected for the lunar dust system due to the large amounts of agglutinic glass and silicate vapor deposits present in lunar soil. Unfortunately, exposure to the Earth s atmosphere has passivated the active species on lunar dust, leading to efforts to reactivate the dust in order to understand the true effects that will be experienced by astronauts and equipment on the moon. Electron spin resonance (ESR) spectroscopy is commonly used for the study of radical species, and has been used previously to study silicon- and oxygen-based radicals, as well as the hydroxyl radicals produced by these species in solution (V. Vallyathan, et al., Am. Rev. Respir. Dis. 138 (1988) 1213-1219). The size and cost of these instruments makes them unattractive for the monitoring of lunar dust activity. A more suitable technique is based on the change in fluorescence of a molecule upon reaction with a hydroxyl radical (or other radical species). Fluorescence instruments are much less costly and bulky than ESR spectrometers, and small fluorescence sensors for space missions have already been developed (F. Gao, et al., J. Biomed. Opt. 10 (2005) 054005). For the current fluorescence studies, the terephthalate molecule has been chosen for monitoring the production of hydroxyl radicals in solution. As shown in Scheme 1, the reaction between the non-fluorescent terephthalate molecule and a hydroxyl radical produces the highly-fluorescent 2-hydroxyterephthalate molecule.

Wallace, W. T.↗

Sensor for infrared communication using plant nanobionics

A living plant can function as self-powered auto-samplers and preconcentrators of an analyte within ambient groundwater, detectors of the analyte contained therein. For example, a pair of near infrared (IR) fluorescent sensors embedded within the mesophyll of the plant leaf can be used as detectors of the nitroaromatic molecules, with the first IR channel engineered through CoPhMoRe to recognize analyte via an IR fluorescent emission and the second IR channel including a functionalized nanostructure that acts as an invariant reference signal.

Strano, Michael S.↗

Airborne Instruments for the In Situ Detection of ClONO2, NO2, ClO, and BrO in the Stratosphere

Design and construction of an in situ sensor for the detection of stratospheric ClONO2, ClO, BrO, and NO2, was conceived as a two-year program. The experiment has two novel components: a resistive silicon thermal dissociation heater used to fragment ClONO2 into ClO and NO2 and a laser-induced fluorescence sensor for N02. These two new components are integrated into an experiment that uses technology developed in our labs for the ER-2 ClO and ER-2 HO(X) instruments. During the first year we reconstructed our laboratory prototypes for ClONO2 and NO2 detection and made substantial improvements in the calibration apparatus. Results from these laboratory experiments have been used to refine the design of the flight instrument. During this year we began the design of all of the long-lead items required to produce a flight instrument: including the design and fabrication of the air flow system used to direct stratospheric air to our halogen sensors, design and prototyping of an aircraft-compatible thermal dissociation heater, and development and test of a new high powered laser system. Finally, we have designed and released for fabrication several subsystems.

Anderson, James G.↗

Barium ion sensing with IPG K + molecular probes

Fluorophores covalently bound to azacrown ether ionophores can be assembled into sensitive turn-on chemosensors. The size specificity and electron-rich nature of the ionophore's binding domain contribute to both selectivity and strong turn-on fluorescence sensing by various mechanisms when properly constructed. Aza-18-crown-6 ethers are quite selective for binding to K + and Ba 2+ , yet the more electron-withdrawing dicationic nature of barium imposes a larger electronic effect on turn-on fluorescent sensors. Barium chemosensors can be important for measuring soluble Ba 2+ in drinking water and have gained recent attention for their potential to enhance the detection of rare events in xenon decay. Here we quantify the capability of three chemosensors, marketed for biologically useful K + sensing, as effective probes for Ba 2+ ions. Here, we present measurements from bulk spectrofluorometry to characterize the system in aqueous solutions and demonstrate the usefulness of these species for low-background single-ion fluorescence microscopy, revealing new candidates for Ba 2+ sensing.

Miller, R. L. [Department of Chemistry and Biochem↗

K-region tetrasubstituted deep-blue pyrene-based luminogens: Visual detection of trace nitroaniline

In recent years, fluorescence sensors have been widely used in explosive detection due to their portability and high efficiency. Here, in the present work, two K-region tetrasubstituted deep-blue pyrene-based luminogens were designed and synthesized by introducing electron-donating groups. The optical properties were investigated by experimental and density functional theory (DFT) methods. Reasonable luminescence efficiency and electron rich characteristic allowed for the possible detection of nitro-explosives. Both luminogens, especially TFPy, exhibit a sensitive response towards nitroanilines (NA), with a limit of detection (LOD) for p-NA as low as 2.75 × 10 –8 M, which is mainly attributed to the electron-donating substituents on the periphery of the pyrene core and the extended π-conjugated structure. These research results establish a low-cost and simple strategy for the detection of trace nitroaniline.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Plastic recognition and electrogenic uniport translocation of 1 st -, 2 nd -, and 3 rd -row transition and post-transition metals by primary-active transmembrane P 1B-2 -type ATPase pumps

Transmembrane P 1B -type ATPase pumps catalyze the extrusion of transition metal ions across cellular lipid membranes to maintain essential cellular metal homeostasis and detoxify toxic metals. Zn(II)-pumps of the P 1B-2 -type subclass, in addition to Zn 2+ , select diverse metals (Pb 2+ , Cd 2+ and Hg 2+ ) at their transmembrane binding site and feature promiscuous metal-dependent ATP hydrolysis in the presence of these metals. Yet, a comprehensive understanding of the transport of these metals, their relative translocation rates, and transport mechanism remain elusive. We developed a platform for the characterization of primary-active Zn(II)-pumps in proteoliposomes to study metal selectivity, translocation events and transport mechanism in real-time, employing a “multi-probe” approach with fluorescent sensors responsive to diverse stimuli (metals, pH and membrane potential). Together with atomic-resolution investigation of cargo selection by X-ray absorption spectroscopy (XAS), we demonstrate that Zn(II)-pumps are electrogenic uniporters that preserve the transport mechanism with 1 st -, 2 nd - and 3 rd -row transition metal substrates. Promiscuous coordination plasticity, guarantees diverse, yet defined, cargo selectivity coupled to their translocation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Fluorescent Detection of Carbon Disulfide by a Highly Emissive and Robust Isoreticular Series of Zr-Based Luminescent Metal Organic Frameworks (LMOFs)

Carbon disulfide (CS 2 ) is a highly volatile neurotoxic species. It is known to cause atherosclerosis and coronary artery disease and contributes significantly to sulfur-based pollutants. Therefore, effective detection and capture of carbon disulfide represents an important aspect of research efforts for the protection of human and environmental health. In this study, we report the synthesis and characterization of two strongly luminescent and robust isoreticular metal organic frameworks (MOFs) Zr 6 (µ 3- O) 4 (OH) 8 (tcbpe) 2 (H 2 O) 4 (here termed 1) and Zr 6 (µ 3- O) 4 (OH) 8 (tcbpe-f) 2 (H 2 O) 4 (here termed 2) and their use as fluorescent sensors for the detection of carbon disulfide. Both MOFs demonstrate a calorimetric bathochromic shift in the optical bandgap and strong luminescence quenching upon exposure to carbon disulfide. The interactions between carbon disulfide and the frameworks are analyzed by in-situ infrared spectroscopy and computational modelling by density functional theory. These results reveal that both the Zr metal node and organic ligand act as the preferential binding sites and interact strongly with carbon disulfide.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A spectral study of a radio-frequency plasma-generated flux of atomic oxygen

The active environment of a radio-frequency (RF) plasma generator, with and without low-pressure oxygen, has been characterized through the identification of emission lines in the spectral region from 250 to 900 nm. The environment is shown to be dependent on the partial pressure of oxygen and the power applied to the RF generator. Atomic oxygen has been found in significant amounts as well as atomic hydrogen and the molecular oxygen species O2((sup 1)Sigma). The only charged species observed was the singly charged molecular ion O2(+). With a polymer specimen in the plasma chamber, carbon monoxide was also observed. The significance of these observations with respect to previous studies using this type of generator to stimulate material degradation in space is discussed. The possibility of using these generators as atomic oxygen sources in the development of oxygen atom fluorescence sensors is explored.

Batten, Carmen E.↗

Characterization of Arcjet Flows Using Laser-Induced Fluorescence

A sensor based on laser-induced fluorescence has been installed at the 20-MW NASA Ames Aerodynamic Heating Facility. The sensor has provided new, quantitative, real-time information about properties of the arcjet flow in the highly dissociated, partially ionized, nonequilibrium regime. Number densities of atomic oxygen, flow velocities, heavy particle translational temperatures, and collisional quenching rates have been measured. These results have been used to test and refine computational models of the arcjet flow. The calculated number densities, translational temperatures, and flow velocities are in moderately good agreement with experiment

Bamford, Douglas J.↗

Hyperspectral Radiative Transfer Modeling to Explore the Combined Retrieval of Biophysical Parameters and Canopy Fluorescence from FLEX - Sentinel-3 Tandem Mission Multi-Sensor Data

The FLuorescence EXplorer (FLEX) satellite mission, selected as ESA's 8th Earth Explorer, has been designed forthe measurement of sun-induced fluorescence (F) spectra emitted by plants. This will be accomplished through amulti-sensor approach by placing it in a common orbit in tandem with the Sentinel-3 (S3) mission, which willhave two optical sensors on board, OLCI (Ocean and Land Colour Instrument) and SLSTR (Sea and Land SurfaceTemperature Radiometer) to complement FLEX. These S3 instruments will be used in combination with theimaging spectrometers on board FLEX to provide data useful for atmospheric correction of FLEX data. However,a fully synergetic approach, i.e. by exploiting the spectral and directional information from all tandem missioninstruments together, is an attractive alternative which is explored in this paper. By employing all combined topof-atmosphere (TOA) spectral radiance data, one can (i) characterize the relevant optical properties of the atmosphere,(ii) retrieve biophysical canopy properties including the associated reflectance anisotropy, and (iii)retrieve a more accurate and consistent canopy F.Regarding retrieval methods, Fraunhofer Line Depth (FLD) and Spectral Fitting (SF) are well-known techniquesapplied to hyperspectral data. Both methods depend on a high spectral resolution and assume aLambertian (isotropic) canopy reflectance. However, most vegetation canopies are non-Lambertian. This impliesthat, in particular when ignoring the anisotropic surface reflection, substantial retrieval errors can occur due tothe interaction between atmospheric absorption bands and surface reflectance anisotropy. In this paper, a novelmethod based on spectral radiative transfer (RT) modeling is proposed, in which coupled RT models are used tosimulate TOA radiance spectra. These are then matched with ‘measured' spectra in order to retrieve surfacefluorescence, along with a suite of biophysical parameters, by model inversion through optimization. By applyingcoupled RT models of the soil-leaf-canopy and the surface-atmosphere systems, TOA radiance spectra canbe simulated for all optical sensors of this tandem mission. In this way, complex effects due to surface reflectanceanisotropy and the spectral sampling by the various instruments, which are difficult to compensate for in the endproducts, are properly taken into account by their incorporation in the forward modeling. Next, by model inversionof TOA radiance data via optimization, the most accurate F retrievals can be achieved in a consistentmanner, along with important canopy level biophysical parameters that may help interpret the F spectrum, suchas chlorophyll content and leaf area index (LAI). The potential of this approach has been explored in a numericalexperiment, and the results are presented in this paper. We find that, with the assumed well-characterized andplausible FLEX/S3 instrument performances, the simultaneous retrieval of biophysical canopy parameters and Fspectra would be possible with a remarkable accuracy, provided the correct atmospheric characterization isavailable.

Verhoef, Wouter↗

Detection Technologies for Reactive Oxygen Species: Fluorescence and Electrochemical Methods and Their Applications

Reactive oxygen species (ROS) have been found in plants, mammals, and natural environmental processes. The presence of ROS in mammals has been linked to the development of severe diseases, such as diabetes, cancer, tumors, and several neurodegenerative conditions. The most common ROS involved in human health are superoxide (O 2 •-), hydrogen peroxide (H 2 O 2 ), and hydroxyl radicals (•OH). Organic and inorganic molecules have been integrated with various methods to detect and monitor ROS for understanding the effect of their presence and concentration on diseases caused by oxidative stress. Among several techniques, fluorescence and electrochemical methods have been recently developed and employed for the detection of ROS. This literature review intends to critically discuss the development of these techniques to date, as well as their application for in vitro and in vivo ROS detection regarding free-radical-related diseases. Moreover, important insights into and further steps for using fluorescence and electrochemical methods in the detection of ROS are presented.

59 BASIC BIOLOGICAL SCIENCES↗

Optical fiber sensors for materials and structures characterization

The final technical report on Optical Fiber Sensors for Materials and Structures Characterization, covering the period August 1990 through August 1991 is presented. Research programs in the following technical areas are described; sapphire optical fiber sensors; vibration analysis using two-mode elliptical core fibers and sensors; extrinsic Fabry-Perot interferometer development; and coatings for fluorescent-based sensor. Research progress in each of these areas was substantial, as evidenced by the technical publications which are included as appendices.

Lindner, D. K.↗

Fluorescence Signatures of Rare Earth Metals during Precipitation in Various Conditions

Fluorescence spectroscopy is a widely used sensor methodology that analyzes light emitted from a compound or element as it decays from an excited state. This technique is very sensitive and selective, which is ideal to characterize analytes at lower limits of detection. Key example targets of significant industry and research interest include rare earth elements (REEs) such as dysprosium (Dy) and europium (Eu). These are widely used in advanced technologies including semiconductors, electric vehicle motors, lasers, and permanent magnets. Identifying new sources and responsible reutilization of REEs is essential, and new approaches to extract and recycle REEs could be notably enhanced through the integration of on-line sensors. The sensors can support faster process design, informed scale-up, and cost-effective deployment. This study covers the initial exploration of applying fluorescence-based on-line monitoring to REEs within a precipitation process. This study demonstrates the successful scale-up of a fluorescence -based sensing approach, from stationary cuvettes and small-volume microfluidic devices to continuous flow systems operating at the bench scale (10-25mL). This work also provides initial insight into the challenges of signal’s effects and utility within a turbid environment. Using a modular design for monitoring flowing solutions in a flow tube, fluorescence can be characterized for a variety of analytical targets. In this study, detection performance parameters between the cuvette and flow tube system were compared. Additionally, the response of Dy during precipitation by sodium bicarbonate in the two measurement designs was explored. This letter represents a starting point to bridge the gap between traditional fluorescence sensor measurements in a cuvette to future developments that explore the ability to integrate fluorescence sensors into extraction and separation processes at industrially relevant scales.

fluorescence↗

RuBisCO activity assays: a simplified biochemical redox approach for in vitro quantification and an RNA sensor approach for in vivo monitoring

Abstract Background Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is the most abundant soluble protein in nature. Extensive studies have been conducted for improving its activity in photosynthesis through approaches like protein engineering. Concurrently, multiple biochemical and radiolabeling assays have been developed for determining its activity. Although these existing assays yield reliable results, they require addition of multiple external components, rendering them less convenient and expensive. Therefore, in this study, we have developed two relatively cheaper, convenient, and easily reproducible assays for quantitative and qualitative estimation of RuBisCO activity. Results We simplified a contemporary NADH based spectrophotometric RuBisCO assay by using cyanobacterial cell lysate as the source for Calvin cycle enzymes. We analyzed the influence of inorganic carbon substrates, CO 2 and NaHCO 3 , and varying protein concentrations on RuBisCO activity. Ribulose-1,5-bisphosphate (RuBP) consumption rates for the cultures grown under 5% CO 2 were 5–7 times higher than the ones grown with 20 mM NaHCO 3 , at different protein concentrations. The difference could be due to the impaired activity of carbonic anhydrase in the cell lysate, which is required for the conversion of HCO 3 − to CO 2 . The highest RuBisCO activity of 2.13 nmol of NAD + / µg of Chl-a/ min was observed with 50 µg of protein and 5% CO 2 . Additionally, we developed a novel RNA-sensor based fluorescence assay that is based on the principle of tracking the kinetics of ATP hydrolysis to ADP during the conversion of 3-phosphoglycerate (3-PG) to 1,3-bisphosphoglycerate (1,3-BPG) in the Calvin cycle. Under in vitro conditions, the fluorometric assay exhibited ~ 3.4-fold slower reaction rate (0.37 min −1 ) than the biochemical assay when using 5% CO 2 . We also confirmed the in vivo application of this assay, where increase in the fluorescence was observed with the recombinant strain of Synechocystis sp. PCC 6803 (SSL142) expressing the ADP-specific RNA sensor, compared to the WT. In addition, SSL142 exhibited three-fold higher fluorescence when supplemented with 20 mM NaHCO 3 as compared to the cells that were grown without NaHCO 3 supplementation. Conclusions Overall, we have developed a simplified biochemical assay for monitoring RuBisCO activity and demonstrated that it can provide reliable results as compared to the prior literature. Furthermore, the biochemical assay using 5% CO 2 (100% relative activity) provided faster RuBP consumption rate compared to the biochemical assay utilizing 20 mM NaHCO 3 (30.70% relative activity) and the in vitro fluorometric assay using 5% CO 2 (29.64% relative activity). Therefore, the absorbance-based biochemical assay using 5% CO 2 or higher would be suitable for in vitro quantification of the RuBisCO activity. On the other hand, the RNA-sensor based in vivo fluorometric assay can be applied for qualitative analysis and be used for high-throughput screening of RuBisCO variants. As RuBisCO is an enzyme shared amongst all the photoautotrophs, the assays developed in this study can easily be extended for analyzing the RuBisCO activities even in microalgae and higher plants.

59 BASIC BIOLOGICAL SCIENCES↗

Incorporation of sensing modalities into de novo designed fluorescence-activating proteins

Through the efforts of many groups, a wide range of fluorescent protein reporters and sensors based on green fluorescent protein and its relatives have been engineered in recent years. Here we explore the incorporation of sensing modalities into de novo designed fluorescence-activating proteins, called mini-fluorescence-activating proteins (mFAPs), that bind and stabilize the fluorescent cis -planar state of the fluorogenic compound DFHBI. We show through further design that the fluorescence intensity and specificity of mFAPs for different chromophores can be tuned, and the fluorescence made sensitive to pH and Ca 2+ for real-time fluorescence reporting. Bipartite split mFAPs enable real-time monitoring of protein–protein association and (unlike widely used split GFP reporter systems) are fully reversible, allowing direct readout of association and dissociation events. The relative ease with which sensing modalities can be incorporated and advantages in smaller size and photostability make de novo designed fluorescence-activating proteins attractive candidates for optical sensor engineering.

59 BASIC BIOLOGICAL SCIENCES↗

A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging

Abstract Genetically encoded fluorescent calcium indicators allow cellular-resolution recording of physiology. However, bright, genetically targetable indicators that can be multiplexed with existing tools in vivo are needed for simultaneous imaging of multiple signals. Here we describe WHaloCaMP, a modular chemigenetic calcium indicator built from bright dye-ligands and protein sensor domains. Fluorescence change in WHaloCaMP results from reversible quenching of the bound dye via a strategically placed tryptophan. WHaloCaMP is compatible with rhodamine dye-ligands that fluoresce from green to near-infrared, including several that efficiently label the brain in animals. When bound to a near-infrared dye-ligand, WHaloCaMP shows a 7× increase in fluorescence intensity and a 2.1-ns increase in fluorescence lifetime upon calcium binding. We use WHaloCaMP1a to image Ca 2+ responses in vivo in flies and mice, to perform three-color multiplexed functional imaging of hundreds of neurons and astrocytes in zebrafish larvae and to quantify Ca 2+ concentration using fluorescence lifetime imaging microscopy (FLIM).

Biochemistry & Molecular Biology↗

Phosphate binding protein as the biorecognition element in a biosensor for phosphate

This work explores the potential use of a member of the periplasmic family of binding proteins, the phosphate binding protein (PBP), as the biorecognition element in a sensing scheme for the detection of inorganic phosphate (Pi). The selectivity of this protein originates from its natural role which, in Escherichia coli, is to serve as the initial receptor for the highly specific translocation of Pi to the cytoplasm. The single polypeptide chain of PBP is folded into two similar domains connected by three short peptide linkages that serve as a hinge. The Pi binding site is located deep within the cleft between the two domains. In the presence of the ligand, the two globular domains engulf the former in a hinge-like manner. The resultant conformational change constitutes the basis of the sensor development. A mutant of PBP (MPBP), where an alanine was replaced by a cysteine residue, was prepared by site-directed mutagenesis using the polymerase chain reaction (PCR). The mutant was expressed, from plasmid pSD501, in the periplasmic space of E. coli and purified in a single chromatographic step on a perfusion anion-exchange column. Site-specific labeling was achieved by attaching the fluorophore, N-[2-(1-maleimidyl)ethyl]-7-(diethylamino)coumarin-3-carboxamide (MDCC), to the protein through the sulfhydryl group of the cysteine moiety. Steady-state fluorescence studies of the MPBP-MDCC conjugate showed a change in the intensity of the signal upon addition of Pi. Calibration curves for Pi were constructed by relating the intensity of the fluorescence signal with the amount of analyte present in the sample. The sensing system was first developed and optimized on a spectrofluorometer using ml volumes of sample. It was then adapted to be used on a microtiter plate arrangement with microliter sample volumes. The system's versatility was finally proven by developing a fiber optic fluorescence-based sensor for monitoring Pi. In all three cases the detection limits for the analyte were in the sub-microMolar range. It was also demonstrated that the sensing system was selective for phosphate over other structurally-similar anions, paving the way for the design and development of a new family of biosensors utilizing the specific binding properties of periplasmic proteins. c2003 Elsevier B.V. All rights reserved.

NASA Program Fundamental Space Biology↗