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At least 703 records · Page 39
All sourcesNASA NTRSDOE OSTI
NASA NTRS2015

Evaluation of the Snow Simulations from the Community Land Model, Version 4 (CLM4)

This paper evaluates the simulation of snow by the Community Land Model, version 4 (CLM4), the land model component of the Community Earth System Model, version 1.0.4 (CESM1.0.4). CLM4 was run in an offline mode forced with the corrected land-only replay of the Modern-Era Retrospective Analysis for Research and Applications (MERRA-Land) and the output was evaluated for the period from January 2001 to January 2011 over the Northern Hemisphere poleward of 30 deg N. Simulated snow-cover fraction (SCF), snow depth, and snow water equivalent (SWE) were compared against a set of observations including the Moderate Resolution Imaging Spectroradiometer (MODIS) SCF, the Interactive Multisensor Snow and Ice Mapping System (IMS) snow cover, the Canadian Meteorological Centre (CMC) daily snow analysis products, snow depth from the National Weather Service Cooperative Observer (COOP) program, and Snowpack Telemetry (SNOTEL) SWE observations. CLM4 SCF was converted into snow-cover extent (SCE) to compare with MODIS SCE. It showed good agreement, with a correlation coefficient of 0.91 and an average bias of -1.54 x 10(exp 2) sq km. Overall, CLM4 agreed well with IMS snow cover, with the percentage of correctly modeled snow-no snow being 94%. CLM4 snow depth and SWE agreed reasonably well with the CMC product, with the average bias (RMSE) of snow depth and SWE being 0.044m (0.19 m) and -0.010m (0.04 m), respectively. CLM4 underestimated SNOTEL SWE and COOP snow depth. This study demonstrates the need to improve the CLM4 snow estimates and constitutes a benchmark against which improvement of the model through data assimilation can be measured.

northern hemisphere↗
NASA NTRS2012

Onboard Science and Applications Algorithm for Hyperspectral Data Reduction

An onboard processing mission concept is under development for a possible Direct Broadcast capability for the HyspIRI mission, a Hyperspectral remote sensing mission under consideration for launch in the next decade. The concept would intelligently spectrally and spatially subsample the data as well as generate science products onboard to enable return of key rapid response science and applications information despite limited downlink bandwidth. This rapid data delivery concept focuses on wildfires and volcanoes as primary applications, but also has applications to vegetation, coastal flooding, dust, and snow/ice applications. Operationally, the HyspIRI team would define a set of spatial regions of interest where specific algorithms would be executed. For example, known coastal areas would have certain products or bands downlinked, ocean areas might have other bands downlinked, and during fire seasons other areas would be processed for active fire detections. Ground operations would automatically generate the mission plans specifying the highest priority tasks executable within onboard computation, setup, and data downlink constraints. The spectral bands of the TIR (thermal infrared) instrument can accurately detect the thermal signature of fires and send down alerts, as well as the thermal and VSWIR (visible to short-wave infrared) data corresponding to the active fires. Active volcanism also produces a distinctive thermal signature that can be detected onboard to enable spatial subsampling. Onboard algorithms and ground-based algorithms suitable for onboard deployment are mature. On HyspIRI, the algorithm would perform a table-driven temperature inversion from several spectral TIR bands, and then trigger downlink of the entire spectrum for each of the hot pixels identified. Ocean and coastal applications include sea surface temperature (using a small spectral subset of TIR data, but requiring considerable ancillary data), and ocean color applications to track biological activity such as harmful algal blooms. Measuring surface water extent to track flooding is another rapid response product leveraging VSWIR spectral information.

Chien, Steve A.↗
NASA NTRS2003

A First Attempt to Simulate the Natural Formation of Meteoritic Organics

Our working hypothesis in these experiments is that meteoritic organics are a mixture of materials made both in the interstellar medium and in the solar nebula that were further processed within meteorite parent bodies to produce what we see today. The material synthesized in the pre-solar environment is currently under study in several laboratories, though to our knowledge none are investigating the changes that might occur as the result of metamorphic processes on the parent body itself. Material made in the Solar Nebula results from a Fischer-Tropsch type (FTT) synthesis as CO and H2 react on grain surfaces to make methane, as well as more complex species. The products of these reactions using natural catalysts have been well-studied and a calculation of the potential efficiency of FTT reactions in the solar nebula demonstrates that they could contribute significantly to the composition of material near 3 AU.

Gardner, Kathryn↗
NASA NTRS2012

Dust Production of Comet 21P/Giacobini-Zinner Using Broadband Photometry

Comet 21P/Giacobini-Zinner is a Jupiter family comet, approximately 2 km in diameter, and is established to be the parent of the Draconids, a meteor shower known to outburst. In 1933 and 1946 up to 10,000 meteors per hour were reported for the Draconids, and 2011 saw a minor Draconid outburst. Meteor stream modeling/forecasting being a primary focus for the NASA Meteoroid Environment Office, it was decided to monitor 21P for three purposes: firstly to find the apparent and absolute magnitude with respect to heliocentric distance; second to calculate Af , a quantity that describes the dust production rate and is used in models to predict the activity of the Draconids; and thirdly to detect possible increases in cometary activity, which could correspond to future Draconid meteor outbursts. A similar study was done for 21P during its 2004-2006 close approach to the Sun in which apparent and absolute magnitudes were found with various heliocentric distances, as well as the dust production. At 2.32 AU from the Sun, 21P possessed an apparent magnitude of 17.05 and Af of 83 cm, and an apparent magnitude of 15.91 and Af of 130.66 cm at 1.76 AU from the sun.

Blaauw, R. C.↗
DOE OSTI2021

Zymomonas diversity and potential for biofuel production

Abstract Background Zymomonas mobilis is an aerotolerant α-proteobacterium, which has been genetically engineered for industrial purposes for decades. However, a comprehensive comparison of existing strains on the genomic level in conjunction with phenotype analysis has yet to be carried out. We here performed whole-genome comparison of 17 strains including nine that were sequenced in this study. We then compared 15 available Zymomonas strains for their natural abilities to perform under conditions relevant to biofuel synthesis. We tested their growth in anaerobic rich media, as well as growth, ethanol production and xylose utilization in lignocellulosic hydrolysate. We additionally compared their tolerance to isobutanol, flocculation characteristics, and ability to uptake foreign DNA by electroporation and conjugation. Results Using clustering based on 99% average nucleotide identity (ANI), we classified 12 strains into four clusters based on sequence similarity, while five strains did not cluster with any other strain. Strains belonging to the same 99% ANI cluster showed similar performance while significant variation was observed between the clusters. Overall, conjugation and electroporation efficiencies were poor across all strains, which was consistent with our finding of coding potential for several DNA defense mechanisms, such as CRISPR and restriction–modification systems, across all genomes. We found that strain ATCC31821 (ZM4) had a more diverse plasmid profile than other strains, possibly leading to the unique phenotypes observed for this strain. ZM4 also showed the highest growth of any strain in both laboratory media and lignocellulosic hydrolysate and was among the top 3 strains for isobutanol tolerance and electroporation and conjugation efficiency. Conclusions Our findings suggest that strain ZM4 has a unique combination of genetic and phenotypic traits that are beneficial for biofuel production and propose investing future efforts in further engineering of ZM4 for industrial purposes rather than exploring new Zymomonas isolates.

09 BIOMASS FUELS↗
DOE OSTI2020

Large-scale production of 88 Y and 88 Zr/ 88 Y generators: A proof of concept study for a 70 MeV H – cyclotron

Here, the large-scale production of 88 Y with proton-induced reactions has been investigated from the perspective of new generation 70 MeV H – cyclotrons. Tandem target configurations are presented for both the direct production of 88 Y as well as for producing 88 Zr/ 88 Y generators. Based on the relevant excitation functions, physical yields have been derived for 88 Y production with Y 2 O 3 /SrCO 3 tandem targets and 88 Zr production with Zr/Y 2 O 3 tandem targets. Yields are presented for optimized targets (i.e. optimum yield) as well as for balanced thermal loads on the individual targets. Liquid 88 Zr/ 88 Y generators have been produced using both natural Zr and Nb target materials, the former for dedicated productions and the latter as a byproduct by processing spent irradiated Nb capsules which normally would constitute radioactive waste. These stock solutions, which contain both the target material and 88 Zr precursor, are retained virtually unchanged after processing except for the removal of 88 Y on AG MP-50 macroporous cation-exchange resin. Methods are presented for the preparation of Nb stock solutions in hydrofluoric acid and Zr stock solutions in sulfuric acid. It is shown that multi-Ci productions of 88 Y are feasible at a 70 MeV cyclotron facility, suitable for the needs of fracking applications. In addition, 88 Zr/ 88 Y generators can provide 88 Y with very high specific activity, suitable for labelling of biomolecules.

07 ISOTOPE AND RADIATION SOURCES↗
DOE OSTI2024

Search for a third-generation leptoquark coupled to a τ lepton and a b quark through single, pair, and nonresonant production in proton-proton collisions at $ \sqrt{s} $ = 13 TeV

A search is presented for a third-generation leptoquark (LQ) coupled exclusively to a τ lepton and a b quark. The search is based on proton-proton collision data at a center-of-mass energy of 13 TeV recorded with the CMS detector, corresponding to an integrated luminosity of 138 fb$^{−1}$. Events with τ leptons and a varying number of jets originating from b quarks are considered, targeting the single and pair production of LQs, as well as nonresonant t-channel LQ exchange. An excess is observed in the data with respect to the background expectation in the combined analysis of all search regions. For a benchmark LQ mass of 2 TeV and an LQ-b-τ coupling strength of 2.5, the excess reaches a local significance of up to 2.8 standard deviations. Upper limits at the 95% confidence level are placed on the LQ production cross section in the LQ mass range 0.5–2.3 TeV, and up to 3 TeV for t-channel LQ exchange. Leptoquarks are excluded below masses of 1.22–1.88 TeV for different LQ models and varying coupling strengths up to 2.5. The study of nonresonant ττ production through t-channel LQ exchange allows lower limits on the LQ mass of up to 2.3 TeV to be obtained.[graphic not available: see fulltext]

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗
DOE OSTI2021

Natural Product Gene Clusters in the Filamentous Nostocales Cyanobacterium HT-58-2

Cyanobacteria are known as rich repositories of natural products. One cyanobacterial-microbial consortium (isolate HT-58-2) is known to produce two fundamentally new classes of natural products: the tetrapyrrole pigments tolyporphins A–R, and the diterpenoid compounds tolypodiol, 6-deoxytolypodiol, and 11-hydroxytolypodiol. The genome (7.85 Mbp) of the Nostocales cyanobacterium HT-58-2 was annotated previously for tetrapyrrole biosynthesis genes, which led to the identification of a putative biosynthetic gene cluster (BGC) for tolyporphins. Here, bioinformatics tools have been employed to annotate the genome more broadly in an effort to identify pathways for the biosynthesis of tolypodiols as well as other natural products. A putative BGC (15 genes) for tolypodiols has been identified. Four BGCs have been identified for the biosynthesis of other natural products. Two BGCs related to nitrogen fixation may be relevant, given the association of nitrogen stress with production of tolyporphins. The results point to the rich biosynthetic capacity of the HT-58-2 cyanobacterium beyond the production of tolyporphins and tolypodiols.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2022

Soil C stock and CO2 production data for Soong et al. 2021: Effects of five years of soil warming at Blodgett Forest, CA.

Subsoils below 20 cm are an important reservoir in the global carbon cycle, but little is known about their vulnerability under climate change. We measured subsoil carbon in warmed vs control plots of a conifer forest after 4.5 years of whole-soil warming (4°C). We also measured soil CO2 efflux at the surface and soil CO2 concentrations along the soil profile. Over the 4.5 years we observed that warming resulted in a loss of subsoil carbon stocks (−33 ± 11%), primarily from unprotected particulate organic matter. Warming also stimulated a sustained 30 ± 4% increase in soil CO2 efflux due to increased CO2 production through the whole-soil profile. The observed in situ decline in subsoil carbon stocks with warming is now definitive evidence of a positive soil carbon-climate feedback, which could not be concluded based on increases in CO2 effluxes alone. This dataset comprises (1) 13C and C recovery in 2014, 2016, and 2018 1m soil cores sampled in 10cm increments. (2) Monthly surface CO2 fluxes from 2014 to 2018. (3) Monthly gas well sample [CO2] and production data. And (4) density fraction data for three depths of cores recovered in 2017.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Modeling suggests that virion production cycles within individual cells is key to understanding acute hepatitis B virus infection kinetics

Hepatitis B virus (HBV) infection kinetics in immunodeficient mice reconstituted with humanized livers from inoculation to steady state is highly dynamic despite the absence of an adaptive immune response. To recapitulate the multiphasic viral kinetic patterns, we developed an agent-based model that includes intracellular virion production cycles reflecting the cyclic nature of each individual virus lifecycle. The model fits the data well predicting an increase in production cycles initially starting with a long production cycle of 1 virion per 20 hours that gradually reaches 1 virion per hour after approximately 3–4 days before virion production increases dramatically to reach to a steady state rate of 4 virions per hour per cell. Together, modeling suggests that it is the cyclic nature of the virus lifecycle combined with an initial slow but increasing rate of HBV production from each cell that plays a role in generating the observed multiphasic HBV kinetic patterns in humanized mice.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1999

The Status of the Tropical Rainfall Measuring Mission (TRMM) after 2 Years in Orbit

The Tropical Rainfall Measuring Mission (TRMM) satellite was launched on November 27, 1997, and data from all the instruments first became available approximately 30 days after launch. Since then, much progress has been made in the calibration of the sensors, the improvement of the rainfall algorithms, in related modeling applications and in new datasets tailored specifically for these applications. This paper reports the latest results regarding the calibration of the TRMM Microwave Imager, (TMI), Precipitation Radar (PR) and Visible and Infrared Sensor (VIRS). For the TMI, a new product is in place that corrects for a still unknown source of radiation leaking in to the TMI receiver. The PR calibration has been adjusted upward slightly (by 0.6 dBZ) to better match ground reference targets, while the VIRS calibration remains largely unchanged. In addition to the instrument calibration, great strides have been made with the rainfall algorithms as well, with the new rainfall products agreeing with each other to within less than 20% over monthly zonally averaged statistics. The TRMM Science Data and Information System (TSDIS) has responded equally well by making a number of new products, including real-time and fine resolution gridded rainfall fields available to the modeling community. The TRMM Ground Validation (GV) program is also responding with improved radar calibration techniques and rainfall algorithms to provide more accurate GV products which will be further enhanced with the new multiparameter 10 cm radar being developed for TRMM validation and precipitation studies. Progress in these various areas has, in turn, led to exciting new developments in the modeling area where Data Assimilation, and Weather Forecast models are showing dramatic improvements after the assimilation of observed rainfall fields.

Kummerow, C.↗
NASA NTRS2010

Re-Engineering Complex Legacy Systems at NASA

The Flight Production Process (FPP) Re-engineering project has established a Model-Based Systems Engineering (MBSE) methodology and the technological infrastructure for the design and development of a reference, product-line architecture as well as an integrated workflow model for the Mission Operations System (MOS) for human space exploration missions at NASA Johnson Space Center. The design and architectural artifacts have been developed based on the expertise and knowledge of numerous Subject Matter Experts (SMEs). The technological infrastructure developed by the FPP Re-engineering project has enabled the structured collection and integration of this knowledge and further provides simulation and analysis capabilities for optimization purposes. A key strength of this strategy has been the judicious combination of COTS products with custom coding. The lean management approach that has led to the success of this project is based on having a strong vision for the whole lifecycle of the project and its progress over time, a goal-based design and development approach, a small team of highly specialized people in areas that are critical to the project, and an interactive approach for infusing new technologies into existing processes. This project, which has had a relatively small amount of funding, is on the cutting edge with respect to the utilization of model-based design and systems engineering. An overarching challenge that was overcome by this project was to convince upper management of the needs and merits of giving up more conventional design methodologies (such as paper-based documents and unwieldy and unstructured flow diagrams and schedules) in favor of advanced model-based systems engineering approaches.

Ruszkowski, James↗
DOE OSTI2021

HFTS-2 Completions Design and State-of-the-Art Diagnostics Results

Various completion schemes were tested as part of the Hydraulic Fracture Test Site-2 (HFTS-2) project. The wells were instrumented with permanent fiber optic (FO) cable, and unique near-wellbore and far-field region surveys were acquired to evaluate spatial and temporal hydraulic fracture characteristics. The state-of-the-art diagnostics included substantial monitoring of both the well stimulation and longer-term production performance. The objective was to evaluate and improve stimulation distribution effectiveness (SDE) for different completion designs based on the diagnostics results. A hydraulic fracturing (HF) profile (i.e., proppant/fluid allocation among clusters) was obtained using near-wellbore Distributed Acoustic Sensing (DAS) data acquired during stimulation. Statistical analysis provided a quantitative assessment of SDE for the different completion designs. A novel workflow was developed to assess time-lapse fracture property changes at the cluster and stage levels based on Distributed Strain Sensing (DSS) data acquired during the production phase. These time-lapse fracture property changes were compared with the HF profile. Based on the statistical analysis, Rate Per Cluster (RPC) during proppant slurry placement appears to be a primary completion design variable, rather than the typical Rate Per Perf (RPP). If RPC was not maintained at a certain level, the likelihood of uneven distribution increased dramatically. The stage configurations that created low RPC while maintaining high RPP did not show improved SDE. The strain change results based on DSS data during the production phase indicated near-wellbore fracture property changes (e.g., aperture changes during well shut-in and flow periods) and aligned well with near-wellbore DAS signals acquired during stimulation. More importantly, fracture property changes based on DSS data acquired during two tests over a 7-month period show high consistency and good alignment with the near-wellbore HF profile. This workflow quantitatively evaluates SDE for different completion designs using diagnostic results and could help optimize completion strategy for future development projects. Here, the analysis and interpretation of results demonstrate the technical feasibility of using permanent fiber optic cable for stimulation and production monitoring and indic

58 GEOSCIENCES↗
DOE OSTI2020

RRNPP-type quorum-sensing systems regulate solvent formation, sporulation and cell motility in Clostridium saccharoperbutylacetonicum

Abstract Background Clostridium saccharoperbutylacetonicum N1-4 (HMT) is a strictly anaerobic, spore-forming Gram-positive bacterium capable of hyper-butanol production through the well-known acetone–butanol–ethanol fermentation process. Recently, five putative RRNPP-type QSSs (here designated as QSS1 to QSS5) were predicted in this bacterial strain, each of which comprises a putative RRNPP-type regulator (QssR1 to QssR5) and a cognate signaling peptide precursor (QssP1 to QssP5). In addition, both proteins are encoded by the same operon. The functions of these multiple RRNPP-type QSSs are unknown. Results To elucidate the function of multiple RRNPP-type QSSs as related to cell metabolism and solvent production in N1-4 (HMT), we constructed qssR -deficient mutants ΔR1, ΔR2, ΔR3 and ΔR5 through gene deletion using CRISPR–Cas9 and N1-4-dcas9-R4 (with the QssR4 expression suppressed using CRISPR–dCas9). We also constructed complementation strains by overexpressing the corresponding regulator gene. Based on systematic characterization, results indicate that QSS1, QSS2, QSS3, and QSS5 positively regulate the sol operon expression and thus solvent production, but they likely negatively regulate cell motility. Consequently, QSS4 might not directly regulate solvent production, but positively affect cell migration. In addition, QSS3 and QSS5 appear to positively regulate sporulation efficiency. Conclusions Our study provides the first insights into the roles of multiple RRNPP-type QSSs of C. saccharoperbutylacetonicum for the regulation of solvent production, cell motility, and sporulation. Results of this study expand our knowledge of how multiple paralogous QSSs are involved in the regulation of essential bacterial metabolism pathways.

09 BIOMASS FUELS↗
DOE OSTI2021

Miscellaneous Electric Loads: Characterization and Energy Savings Potential

Over time, miscellaneous electric loads (MELs) are expected to increase both in magnitude and share of residential and commercial building energy consumption. This trend is most apparent in North America, but it is also occurring in Japan and Europe. However, the contribution of MELs to building energy use is not currently well understood, both because the products in this category are transforming rapidly and the definition and classification of MELs is ambiguous. This study estimated the national energy consumption of 36 MELs using best-available data and found them to comprise 12% of delivered electricity to the U.S. residential and commercial building sectors. If 26 of these MELs were replaced with the most energy-efficient product models available on the market, their energy consumption could be halved to 6% of delivered electricity. National energy models will better account for building energy consumption by incorporating the MELs data collected and analyzed for this study, leading to improved policy decisions.

Miscellaneous electric loads, Plug loads, Taxonomy↗
NASA NTRS2002

Plant Growth During the Greenhouse II Experiment on the MIR Orbital Station

We carried out three experiments with Super Dwarf wheat in the Bulgarian/Russian growth chamber Svet (0.1 sq m growing area) on the Space Station Mir. This paper mostly describes the first of these NASA-supported trials, began on Aug. 13, 1995. Plants were sampled five times and harvested on Nov. 9 after 90 days. Equipment failures led to low irradiance (three, then four of six lamp sets failed), instances of high temperatures (ca. 37 C), and sometimes excessive-substrate moisture. Although plants grew for the 90 days, no wheat heads were produced. Considering the low light levels, plants were surprisingly green, but of course biomass production was low. Plants were highly disoriented (low light, mirror walls?). Fixed and dried samples and the root module were returned on the US Shuttle Atlantis on Nov. 20, 1995. Samples of the substrate, a nutrient-charged zeolite called Balkanine, were taken from the root module, carefully examined for roots, weighed, dried, and reweighed. The Svet control unit and the light bank were shipped to Moscow. An experiment validation test (EVT) of plant growth and experiment procedures, carried out in Moscow, was highly successful. Equipment built in Utah to measure CO2, H2O vapor, irradiance, air and leaf (IR) temperature, O2, pressure, and substrate moisture worked well in the EVT and in space. After this manuscript was first prepared, plants were grown in Mir with a new light bank and controller for 123 days in late 1996 and 39 days in 1996/1997. Plants grew exceptionally well with higher biomass production than in any previous space experiment, but the ca. 280 wheat heads that were produced in 1996 contained no seeds. Ethylene in the cabin atmosphere was responsible.

Salisbury, F. B.↗
NASA NTRS2003

Plant growth during the Greenhouse II experiment on the Mir orbital station

We carried out three experiments with Super Dwarf wheat in the Bulgarian/Russian growth chamber Svet (0.1 m2 growing area) on the Space Station Mir. This paper mostly describes the first of these NASA-supported trials, began on Aug. 13, 1995. Plants were sampled five times and harvested on Nov. 9 after 90 days. Equipment failures led to low irradiance (3, then 4 of 6 lamp sets failed), instances of high temperatures (ca. 37 degrees C), and sometimes excessive substrate moisture. Although plants grew for the 90 d, no wheat heads were produced. Considering the low light levels, plants were surprisingly green, but of course biomass production was low. Plants were highly disoriented (low light, mirror walls?). Fixed and dried samples and the root module were returned on the U.S. Shuttle Atlantis on Nov. 20, 1995. Samples of the substrate, a nutrient-charged zeolite called Balkanine, were taken from the root module, carefully examined for roots, weighed, dried, and reweighed. The Svet control unit and the light bank were shipped to Moscow. An experiment validation test (EVT) of plant growth and experimental procedures, carried out in Moscow, was highly successful. Equipment built in Utah to measure CO2, H2O vapor, irradiance, air and leaf (IR) temperature, O2, pressure, and substrate moisture worked well in the EVT and in space. After this manuscript was first prepared, plants were grown in Mir with a new light bank and controller for 123 d in late 1996 and 39 days in 1996/1997. Plants grew exceptionally well with higher biomass production than in any previous space experiment, but the ca. 280 wheat heads that were produced in 1996 contained no seeds. Ethylene in the cabin atmosphere was responsible. c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

Mir Project↗
DOE OSTI2023

Development and implementation of an in situ high-resolution isotopic microscope for measuring metabolic interactions in soil mesocosms (Final Report)

Many of our planet’s ecosystems rely on the activities of soil microbial communities. These microbes have significant and wide-ranging effects: they metabolize carbon and other nutrients, interact with plants and fungi, and perform other processes important to soil health. However, our ability to directly observe the enzymatic and metabolic activities of microbes within soil is currently limited, not only by the complexity of soil microbial communities themselves, but also by the lack of experimental tools to study them and their molecular interactions in situ. These challenges hinder our understanding of the life-sustaining processes of biomass decomposition and the manner in which it contributes to the movement of freed carbon within soil ecosystems. Here we aimed to develop a novel ultrahigh-resolution isotopic microscope that combines complementary imaging modalities to gain insights into metabolic cycling in soil. This final report covers both the portion of this work that was initially completed at the University of North Carolina at Chapel Hill from 2018 to 2019 (as award DE-SC0019012) and then from 2020 to 2022 (2023 in NCE) at the University of Massachusetts Chan Medical School. This goal was to build an integrated platform consisting of fluorescence microscopy, Raman microspectroscopy, and nanospray desorption electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (nanoDESI-FTICR-MS) to directly investigate microbial activities and molecular transformations occurring in soil by exploiting the use of both fluorescent labels and stable isotope probing. Specifically, our instrument was conceived as integrating: (A) fluorescence detection to localize soil microbes, identify bacteria taking up polysaccharides, and monitor gene expression of enzymes involved in decomposition; (B) Raman microspectroscopy to determine which microbes incorporate decomposition products into their biomass, as well as which decomposers are metabolically active; and (C) nanoDESI-FTICR-MS imaging to spatially probe, in real-time, the metabolites in the surrounding area, which we expect will reveal the distributions of the products of the enzymatic breakdown of polysaccharides, as well as specialized metabolites acting as cell-cell signals between decomposers. To enable these measurements, we exploited fluorescence-based probes to map the microbes that are enzymatically active (per A) and utilized stable isotope-labeled substrates to visualize both the microbial and molecular fate of decomposed biomass (B and C, respectively). To accomplish our goal, we combined the expertise of a multidisciplinary group of scientists to pursue the construction of this microscope and to investigate scientific questions that would be facilitated by such a capability. We aimed to employ this spatially informative, high-resolution isotopic microscope to visualize the critical steps of biomass degradation and the molecular fate of other environmentally relevant substrates within soil mesocosms. Accordingly, this technology will enhance our understanding of the microbial and metabolic interactions occurring within soil communities that are relevant to carbon degradation and other soil processes. The instrument has been developed and housed at EMSL, where it will be available to the entire EMSL User Base, enabling a variety of related DOE-relevant systems to be interrogated in the future by diverse scientific research groups.

59 BASIC BIOLOGICAL SCIENCES↗