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NASA NTRS2001

Distribution and Spectroscopy of Green Fluorescent Protein and Acyl-CoA: Cholesterol Acytransferase in Sf21 Insect Cells

Acyl-CoA: cholesterol acyltransferase (ACAT) is thought to significantly participate in the pathway of cholesterol esterification that underlies the pathology of artherosclerosis. This enzyme is a membrane protein known to be preferentially bound within the endoplasmic reticulum of mammalian cells, from which location it esterifies cholesterol derived from low density lipoprotein. Cultures of insect cells were separately infected with baculovirus containing the gene for green fluroescent protein (GFP) and with baculovirus containing tandem genes for GFP and ACAT. These infected cultures expressed GFP and the fusion protein GCAT, respectively, with maximum expression occurring on the fourth day after infection. Extraction of GFP- and of GCAT-expressing cells with urea and detergent resulted in recovery of fluorescent protein in aqueous solution. Fluorescence spectra at neutral pH were identical for both GFP and GCAT extracts in aqueous solution, indicating unperturbed tertiary structure for the GFP moiety within GCAT. In a cholesterol esterification assay, GCAT demonstrated ACAT activity, but with less efficiency compared to native ACAT. It was hypothesized that the membrane protein ACAT would lead to differences in localization of GCAT compared to GFP within the respective expressing insect cells. The GFP marker directly and also within the fusion protein GCAT was accordingly used as the intracellular probe that was fluorescently analyzed by the new biophotonics technique of hyperspectral imaging. In that technique, fluorescence imaging was obtained from two dimensional arrays of cells, and regions of interest from within those images were then retrospectively analyzed for the emission spectra that comprises the image. Results of hyperspectral imaging of insect cells on day 4 postinfection showed that GCAT was preferentially localized to the cytoplasm of these cells compared to GFP. Furthermore, the emission spectra obtained for the localized GCAT displayed a peak blue shift from 518nm obtained in neutral aqueous solution to 505nm obtained in localized regions within the cells. This blue shift indicates change in the fluorescence coupling of the GFP moiety of GCAT. It is hypothesized that change in tertiary environment of GCAT, coincident with intracellular deposition of GCAT, follows from intracellular trafficking of GCAT leading to membrane interactions with the ACAT moiety, and/or self-assembly of GCAT, that alters the chromophore environment of the GFP moiety of GCAT. These findings introduce a new technique of biophotonic imaging to studies of intracellular protein trafficking and interactions. This technique of hyperspectral imaging could contribute to advancing the emergent field of proteomics. Because of the noninvasive nature of this technique, kinetic processes associated with intracellular protein trafficking, and interactions of proteins within cellular domains, can be considered for investigation within a single cell as well as a cell population.

Richmond, R. C.↗

NASA NTRS2012

Hyperspectral Feature Detection Onboard the Earth Observing One Spacecraft using Superpixel Segmentation and Endmember Extraction

We present a demonstration of onboard hyperspectral image processing with the potential to reduce mission downlink requirements. The system detects spectral endmembers and then uses them to map units of surface material. This summarizes the content of the scene, reveals spectral anomalies warranting fast response, and reduces data volume by two orders of magnitude. We have integrated this system into the Autonomous Science craft Experiment for operational use onboard the Earth Observing One (EO-1) Spacecraft. The system does not require prior knowledge about spectra of interest. We report on a series of trial overflights in which identical spacecraft commands are effective for autonomous spectral discovery and mapping for varied target features, scenes and imaging conditions.

Superpixel Endmember Detection↗

NASA NTRS2023

The Mars Atmosphere Water Ice Aerosol Climatology by MRO/CRISM: 5 Mars Years of Observations

We use near-infrared spectra returned from the Compact Reconnaissance Imaging Spectrometer for Mars (CRISM) aboard the Mars Reconnaissance Orbiter (MRO) to provide retrievals of the column-integrated optical depth of water ice aerosols for more than 5 Martian years between Mars Year (MY) 28 at Ls = 112° (27 September 2006) and MY 33 at Ls = 213° (30 August 2016). We have developed a radiative transfer model for this purpose that retrieves aerosol opacity from the water ice aerosol feature near 3.3 μm using CRISM hyperspectral observations for the first time. The resulting retrievals well depict the main features in the water ice aerosol climatology with repeatable patterns every Martian year. The aphelion cloud belt (ACB) is observed between 10°S and 30°N, with peak optical depth around Ls = 90°. Apart from the global dust storm during MY 28, modest interannual variability in the water ice aerosol optical depth is observed in the ACB. The north polar hood is observed at latitudes poleward of 45°N throughout northern spring and summer seasons and reappears in northern winter. The south polar hood is observed in early to mid-southern autumn, reaching latitudes around 60°S, and re-emerging during southern winter between 45°S and 60°S. A lack of water ice aerosols is repeatedly seen in the southern hemisphere throughout southern spring and summer. High optical depth in the ACB is observed over the volcanoes of Olympus Mons, Elysium, and the Tharsis bulge, as well as over Hellas Basin.

Mars↗

NASA NTRS2007

Mosaic-Detector-Based Fluorescence Spectral Imager

A battery-powered, pen-sized, portable instrument for measuring molecular fluorescence spectra of chemical and biological samples in the field has been proposed. Molecular fluorescence spectroscopy is among the techniques used most frequently in laboratories to analyze compositions of chemical and biological samples. Heretofore, it has been possible to measure fluorescence spectra of molecular species at relative concentrations as low as parts per billion (ppb), with a few nm spectral resolution. The proposed instrument would include a planar array (mosaic) of detectors, onto which a fluorescence spectrum would be spatially mapped. Unlike in the larger laboratory-type molecular fluorescence spectrometers, mapping of wavelengths to spatial positions would be accomplished without use of relatively bulky optical parts. The proposed instrument is expected to be sensitive enough to enable measurement of spectra of chemical species at relative concentrations <1 ppb, with spectral resolution that could be tailored by design to be comparable to a laboratory molecular fluorescence spectrometer. The proposed instrument (see figure) would include a button-cell battery and a laser diode, which would generate the monochromatic ultraviolet light needed to excite fluorescence in a sample. The sample would be held in a cell bounded by far-ultraviolet-transparent quartz or optical glass. The detector array would be, more specifically, a complementary metal oxide/ semiconductor or charge-coupled- device imaging photodetector array, the photodetectors of which would be tailored to respond to light in the wavelength range of the fluorescence spectrum to be measured. The light-input face of the photodetector array would be covered with a matching checkerboard array of multilayer thin film interference filters, such that each pixel in the array would be sensitive only to light in a spectral band narrow enough so as not to overlap significantly with the band of an adjacent pixel. The wavelength interval between adjacent pixels (and, thus, the spectral resolution) would typically be chosen by design to be approximately equal to the width of the total fluorescence wavelength range of interest divided by the number of pixels. The unitary structure comprising the photodetector array overlaid with the matching filter array would be denoted a hyperspectral mosaic detector (HMD) array.

Son, Kyung-Ah↗

NASA NTRS2011

High Speed, Multi-Channel, Thermal Instrument Development in Support of HyspIRI-TIR

The Jet Propulsion Laboratory is currently developing an end-to-end instrument which will provide a proof of concept prototype vehicle for a high data rate, multi-channel, thermal instrument in support of the Hyperspectral Infrared Imager (HyspIRI)-Thermal Infrared (TIR) space mission. HyspIRI mission was recommended by the National Research Council Decadal Survey (DS). The HyspIRI mission includes a visible shortwave infrared (SWIR) pushboom spectrometer and a multispectral whiskbroom thermal infrared (TIR) imager. The prototype testbed instrument addressed in this effort will only support the TIR. Data from the HyspIRI mission will be used to address key science questions related to the Solid Earth and Carbon Cycle and Ecosystems focus areas of the NASA Science Mission Directorate. Current designs for the HyspIRI-TIR space borne imager utilize eight spectral bands delineated with filters. The system will have 60m ground resolution, 200mK NEDT, 0.5C absolute temperature resolution with a 5-day repeat from LEO orbit. The prototype instrument will use mercury cadmium telluride (MCT) technology at the focal plane array in time delay integration mode. A custom read out integrated circuit (ROIC) will provide the high speed readout hence high data rates needed for the 5 day repeat. The current HyspIRI requirements dictate a ground knowledge measurement of 30m, so the prototype instrument will tackle this problem with a newly developed interferometeric metrology system. This will provide an absolute measurement of the scanning mirror to an order of magnitude better than conventional optical encoders. This will minimize the reliance on ground control points hence minimizing postprocessing (e.g. geo-rectification computations).

Multi-spectral↗

NASA NTRS2015

Crosscutting Airborne Remote Sensing Technologies for Oil and Gas and Earth Science Applications

Airborne imaging spectroscopy has evolved dramatically since the 1980s as a robust remote sensing technique used to generate 2-dimensional maps of surface properties over large spatial areas. Traditional applications for passive airborne imaging spectroscopy include interrogation of surface composition, such as mapping of vegetation diversity and surface geological composition. Two recent applications are particularly relevant to the needs of both the oil and gas as well as government sectors: quantification of surficial hydrocarbon thickness in aquatic environments and mapping atmospheric greenhouse gas components. These techniques provide valuable capabilities for petroleum seepage in addition to detection and quantification of fugitive emissions. New empirical data that provides insight into the source strength of anthropogenic methane will be reviewed, with particular emphasis on the evolving constraints enabled by new methane remote sensing techniques. Contemporary studies attribute high-strength point sources as significantly contributing to the national methane inventory and underscore the need for high performance remote sensing technologies that provide quantitative leak detection. Imaging sensors that map spatial distributions of methane anomalies provide effective techniques to detect, localize, and quantify fugitive leaks. Airborne remote sensing instruments provide the unique combination of high spatial resolution (<1 m) and large coverage required to directly attribute methane emissions to individual emission sources. This capability cannot currently be achieved using spaceborne sensors. In this study, results from recent NASA remote sensing field experiments focused on point-source leak detection, will be highlighted. This includes existing quantitative capabilities for oil and methane using state-of-the-art airborne remote sensing instruments. While these capabilities are of interest to NASA for assessment of environmental impact and global climate change, industry similarly seeks to detect and localize leaks of both oil and methane across operating fields. In some cases, higher sensitivities desired for upstream and downstream applications can only be provided by new airborne remote sensing instruments tailored specifically for a given application. There exists a unique opportunity for alignment of efforts between commercial and government sectors to advance the next generation of instruments to provide more sensitive leak detection capabilities, including those for quantitative source strength determination.

hyperspectral↗