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At least 181 records · Page 10

Control of valley polarization in gapped graphene by linearly polarized ultrashort optical pulse

We present a versatile model of gapped graphene to manipulate valley polarization by using ultrafast linearly polarized pulses of zero areas. First, a circularly polarized pulse of a single oscillation produces a valley-selective population of the conduction band by means of topological resonance. Then we apply a linearly polarized femtosecond-long pulse that changes the valley polarization. The magnitude of such a change depends on the amplitude and the direction of polarization of the pulse. In conclusion, our protocol provides a favorable platform for applications in valleytronics.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Rapid characterization and failure analysis of 6276 rooftop-harvested photovoltaic connectors

Photovoltaic (PV) connectors, which link modules in series and connect PV strings in parallel, have increasingly been recognized as a primary contributor to PV system failures and a source of numerous fire incidents. However, publicly available data on the rates and types of connector failures are scarce, primarily due to the proprietary nature of the information and the need for comprehensive analysis. This study represents the first large-scale investigation of harvested PV connectors, drawing from a dataset of 6276 connectors from residential rooftop solar systems across the United States. The outcome of this work is twofold: 1) we have established a rapid characterization method for large populations of harvested connectors, incorporating visual inspection, resistance measurements, and X-ray imaging; and 2) the analysis made possible by our rapid-processing method has revealed, for a population of connector models provided by a single rooftop installer, failure statistics and insights for various connector makes and models, installation practices, operating currents, and internal component displacements. This research identifies common failure modes that could be considered in future connector designs standards, and operations and maintenance practices, to ultimately improve the reliability of this vital component of PV infrastructure.

MC4↗

Single-cell proteomics of Arabidopsis leaf mesophyll reveals dynamic protein responses to water-deficit stress

Background The application of single-cell omics tools to biological systems can provide unique insights into diverse cellular populations and their heterogeneous responses to internal and external perturbations. Thus far, most single-cell studies in plant systems have been limited to RNA-sequencing approaches, which only provide indirect readouts of cellular functions. Results Here, we present a single-cell proteomics workflow for plant cells that integrates tape-sandwich protoplasting, piezoelectric cell sorting, nanoPOTS sample preparation, and ion mobility-based MS data acquisition method for label-free single-cell proteomics analysis of Arabidopsis leaf mesophyll cells. From a single leaf protoplast, over 3,000 proteins were quantified with high precision. The workflow is demonstrated to identify stress associated changes in protein abundance by analyzing 117 protoplasts from well-watered and water-deficit stressed plants. Additionally, we describe a new approach for constructing covarying protein networks at the single-cell level and demonstrate how single-cell protein covariation analysis can reveal previously unrecognized protein functions while also capturing stress-induced changes in protein–protein dynamics. Conclusions The label-free scProteomic approach presented here represents a significant advance through the demonstration of a facile protoplast isolation method combined with deep and precise proteomic coverage of Arabidopsis leaf mesophyll cell types. We believe this study will serve as an informative reference to future plant scProteomic investigations.

Arabidopsis↗

Evolutionary consequences of nascent multicellular life cycles

A key step in the evolutionary transition to multicellularity is the origin of multicellular groups as biological individuals capable of adaptation. Comparative work, supported by theory, suggests clonal development should facilitate this transition, although this hypothesis has never been tested in a single model system. We evolved 20 replicate populations of otherwise isogenic clonally reproducing ‘snowflake’ yeast (Δace2/∆ace2) and aggregative ‘floc’ yeast (GAL1p::FLO1 /GAL1p::FLO1) with daily selection for rapid growth in liquid media, which favors faster cell division, followed by selection for rapid sedimentation, which favors larger multicellular groups. While both genotypes adapted to this regime, growing faster and having higher survival during the group-selection phase, there was a stark difference in evolutionary dynamics. Aggregative floc yeast obtained nearly all their increased fitness from faster growth, not improved group survival; indicating that selection acted primarily at the level of cells. In contrast, clonal snowflake yeast mainly benefited from higher group-dependent fitness, indicating a shift in the level of Darwinian individuality from cells to groups. Through genome sequencing and mathematical modeling, we show that the genetic bottlenecks in a clonal life cycle also drive much higher rates of genetic drift—a result with complex implications for this evolutionary transition. Our results highlight the central role that early multicellular life cycles play in the process of multicellular adaptation.

Life Sciences & Biomedicine - Other Topics↗

Signal dispersion within a hippocampal neural network

A model network is described, representing two neural populations coupled so that one population is inhibited by activity it excites in the other. Parameters and operations within the model represent EPSPs, IPSPs, neural thresholds, conduction delays, background activity and spatial and temporal dispersion of signals passing from one population to the other. Simulations of single-shock and pulse-train driving of the network are presented for various parameter values. Neuronal events from 100 to 300 msec following stimulation are given special consideration in model calculations.

Horowitz, J. M.↗

Optical characterization of the oceanic unicellular cyanobacterium Synechococcus grown under a day-night cycle in natural irradiance

The optical properties of the ocenanic cyanobacterium Synechococcus (clone WH8103) were examined in a nutrient-replete laboratory culture grown under a day-night cycle in natural irradiance. Measurements of the spectral absorption and beam attenuation coefficients, the size distribution of cells in suspension, and microscopic analysis of samples were made at intervals of 2-4 hours for 2 days. These measurements were used to calculate the optical properties at the level of a single 'mean' cell representative of the acutal population, specifically, the optical cross sections for spectral absorption bar-(sigma(sub a)), scattering bar-sigma(sub b))(lambda), and attentuation bar-(sigma(sub c))(lambda). In addition, concurrent determinations of chlorophyll a and particulate organic carbon allowed calculation of the Chl a- and C-specific optical coefficients. The refractive index of cells was derived from the observed data using a theory of light absorption and scattering by homogeneous spheres. Low irradiance because of cloudy skies resulted in slow division rates of cells in the culture. The percentage of dividing cells was unusually high (greater than 30%) throughout the experiment. The optical cross sections varied greatly over a day-night cycle, with a minimum near dawn or midmorning and maximum near dusk. During daylight hours, bar-(sigma(sub b)) and bar-(sigma(sub c)) can increase more than twofold and bar-(sigma(sub a) by as much as 45%. The real part of the refractive index n increaed during the day; changes in n had equal or greater effect than the varying size distribution on changes in bar-(sigma(sub c)) and bar-(sigma(sub b)). The contribution of changes in n to the increase of bar-(sigma(sub c))(660) during daylight hours was 65.7% and 45.1% on day 1 and 2, respectively. During the dark period, when bar-(sigma(sub c))(660) decreased by a factor of 2.9, the effect of decreasing n was dominant (86.3%). With the exception of a few hours during the second light period, the imaginary part of the refractive index n' showed little variation over a day-night cycle, and bar-(sigma(sub a)) was largely controlled by variations in cell size. The real part of the refractive index at lambda = 660 nm was correlated with the intracellular C concentration and the imaginary part at lambda = 678 nm with the intracellular Chl a concentration. The C-specfic attenuation coefficient showed significant diel variability, which has implications for the estimation of oceanic primary production from measurements of diel variability in beam attenuation. This study provides strong evidence that diel variability is an important component of the optical characterization of marine phytoplankton.

Stramski, Dariusz↗

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.↗

Chromosome Aberrations in Astronauts

A review of currently available data on in vivo induced chromosome damage in the blood lymphocytes of astronauts proves that, after protracted exposure of a few months or more to space radiation, cytogenetic biodosimetry analyses of blood collected within a week or two of return from space provides a reliable estimate of equivalent radiation dose and risk. Recent studies indicate that biodosimetry estimates from single spaceflights lie within the range expected from physical dosimetry and biophysical models, but very large uncertainties are associated with single individual measurements and the total sample population remains low. Retrospective doses may be more difficult to estimate because of the fairly rapid time-dependent loss of "stable" aberrations in blood lymphocytes. Also, biodosimetry estimates from individuals who participate in multiple missions, or very long (interplanetary) missions, may be complicated by an adaptive response to space radiation and/or changes in lymphocyte survival and repopulation. A discussion of published data is presented and specific issues related to space radiation biodosimetry protocols are discussed.

George, Kerry A.↗

Ecological Impacts of the Space Shuttle Program at John F. Kennedy Space Center, Florida

The Space Shuttle Program was one of NASAs first major undertakings to fall under the environmental impact analysis and documentation requirements of the National Environmental Policy Act of 1969 (NEPA). Space Shuttle Program activities at John F. Kennedy Space Center (KSC) and the associated Merritt Island National Wildlife Refuge (MINWR) contributed directly and indirectly to both negative and positive ecological trends in the region through the long-term, stable expenditure of resources over the 40 year program life cycle. These expenditures provided support to regional growth and development in conjunction with other sources that altered land use patterns, eliminated and modified habitats, and contributed to cultural eutrophication of the Indian River Lagoon. At KSC, most Space Shuttle Program related actions were conducted in previously developed facilities and industrial areas with the exception of the construction of the shuttle landing facility (SLF) and the space station processing facility (SSPF). Launch and operations impacts were minimal as a result of the low annual launch rate. The majority of concerns identified during the NEPA process such as potential weather modification, acid rain off site, and local climate change did not occur. Launch impacts from deposition of HCl and particulates were assimilated as a result of the high buffering capacity of the system and low launch and loading rates. Metals deposition from exhaust deposition did not display acute impacts. Sub-lethal effects are being investigated as part of the Resource Conservation and Recovery Act (RCRA) regulatory process. Major positive Space Shuttle Program effects were derived from the adequate resources available at the Center to implement the numerous environmental laws and regulations designed to enhance the quality of the environment and minimize impacts from human activities. This included reduced discharges of domestic and industrial wastewater, creation of stormwater management systems, remediation of past contamination sites, implementation of hazardous waste management systems, and creation of a culture of sustainability. Working with partners such as the USFWS and the St Johns River Water Management District (SJRWMD), wetlands and scrub restoration and management initiatives were implemented to enhance fish and wildlife populations at the Center. KSC remains the single largest preserve on the east coast of Florida in part due to NASAs commitment to stewardship. Ongoing Ecological Program projects are directed at development of information and knowledge to address future KSC management questions including the transition to a joint government and commercial launch facility, enhanced habitat management requirements for wetlands and scrub, potential impacts of emerging contaminants, and adaptation to climate change including projected sea level rise over the next 50-75 years.

Space Shuttle Program↗

Single-cell and spatiotemporal transcriptomic profiling of brain immune infiltration following Venezuelan equine encephalitis virus infection

Neurotropic alphaviruses such as Venezuelan equine encephalitis virus (VEEV) are critical human pathogens that continually expand to naïve populations and for which there are no licensed vaccines or therapeutics. VEEV is highly infectious via the aerosol route and is a recognized weaponizable biothreat that causes neurological disease in humans. The neuropathology of VEEV has been attributed to an inflammatory immune response in the brain yet the underlying mechanisms and specific immune cell populations involved are not fully elucidated. This study uses single-cell RNA sequencing to produce a comprehensive transcriptional profile of immune cells isolated from the brain over a time course of infection in a mouse model of VEEV. Analyses reveal differentially activated subpopulations of microglia, including a distinct type I interferon-expressing subpopulation. This is followed by the sequential infiltration of myeloid cells and cytotoxic lymphocytes, also comprising subpopulations with unique transcriptional signatures. We identify a subpopulation of myeloid cells that form a distinct localization pattern in the hippocampal region whereas lymphocytes are widely distributed, indicating differential modes of recruitment, including that to specific regions of the brain. Altogether, this study provides a high-resolution analysis of the immune response to VEEV in the brain and highlights potential avenues of investigation for therapeutics that target neuroinflammation in the brain.

59 BASIC BIOLOGICAL SCIENCES↗

Survival of endospores of Bacillus subtilis on spacecraft surfaces under simulated martian environments: implications for the forward contamination of Mars

Experiments were conducted in a Mars simulation chamber (MSC) to characterize the survival of endospores of Bacillus subtilis under high UV irradiation and simulated martian conditions. The MSC was used to create Mars surface environments in which pressure (8.5 mb), temperature (-80, -40, -10, or +23 degrees C), gas composition (Earth-normal N2/O2 mix, pure N2, pure CO2, or a Mars gas mix), and UV-VIS-NIR fluence rates (200-1200 nm) were maintained within tight limits. The Mars gas mix was composed of CO2 (95.3%), N2 (2.7%), Ar (1.7%), O2 (0.2%), and water vapor (0.03%). Experiments were conducted to measure the effects of pressure, gas composition, and temperature alone or in combination with Mars-normal UV-VIS-NIR light environments. Endospores of B. subtilis, were deposited on aluminum coupons as monolayers in which the average density applied to coupons was 2.47 x 10(6) bacteria per sample. Populations of B. subtilis placed on aluminum coupons and subjected to an Earth-normal temperature (23 degrees C), pressure (1013 mb), and gas mix (normal N2/O2 ratio) but illuminated with a Mars-normal UV-VIS-NIR spectrum were reduced by over 99.9% after 30 sec exposure to Mars-normal UV fluence rates. However, it required at least 15 min of Mars-normal UV exposure to reduce bacterial populations on aluminum coupons to non-recoverable levels. These results were duplicated when bacteria were exposed to Mars-normal environments of temperature (-10 degrees C), pressure (8.5 mb), gas composition (pure CO2), and UV fluence rates. In other experiments, results indicated that the gas composition of the atmosphere and the temperature of the bacterial monolayers at the time of Mars UV exposure had no effects on the survival of bacterial endospores. But Mars-normal pressures (8.5 mb) were found to reduce survival by approximately 20-35% compared to Earth-normal pressures (1013 mb). The primary implications of these results are (a) that greater than 99.9% of bacterial populations on sun-exposed surfaces of spacecraft are likely to be inactivated within a few tens of seconds to a few minutes on the surface of Mars, and (b) that within a single Mars day under clear-sky conditions bacterial populations on sun-exposed surfaces of spacecraft will be sterilized. Furthermore, these results suggest that the high UV fluence rates on the martian surface can be an important resource in minimizing the forward contamination of Mars. c2003 Elsevier Inc. All rights reserved.

Bacillus subtilis/growth & development/physiology/↗

Single cell RNA sequencing reveals shifts in cell maturity and function of endogenous and infiltrating cell types in response to acute intervertebral disc injury

Intervertebral disc (IVD) degeneration contributes to disabling back pain. Degeneration can be initiated by injury and progressively leads to an irreversible loss of cells and function. IVD function restoration through cell replacement therapies have had limited success due to knowledge gaps in the critical cell populations important for repair. Here, in this study, we used single cell RNA sequencing to identify the transcriptional changes of IVD resident and infiltrating cell populations from Control and Injured coccygeal IVDs extracted from 12-week-old female C57BL/6J mice 7 days post injury. Clustering, gene ontology, and pseudotime trajectory analyses determined transcriptomic divergences with injury, flow cytometry identified they types of infiltrating immune cells, and immunofluorescence was utilized to define mesenchymal stem cell (MSC) localization. We identified 11 distinct clusters that included IVD, immune, vascular cells, and MSCs. Differential gene expression analysis determined that Outer Annulus Fibrosus, Neutrophils, Saa2-High MSCs, Macrophages, and Krt18 + Nucleus Pulposus (NP) cells were the major drivers of transcriptomic differences between Control and Injured cells. Gene ontology revealed that the most upregulated biological pathways were angiogenesis and T cell-related while wound healing and ECM regulation were downregulated. Pseudotime trajectory analyses revealed that IVD injury directed cells towards increased differentiation in all clusters, except for Krt18 + NP cells which remained in a less mature cell state. Saa2-High and Grem1-High MSCs populations shifted towards more differentiated IVD cells profiles with injury and localized distinctly within the IVD. This study revealed novel MSC populations with the potential to be leveraged for future IVD repair studies.

Cartilage↗

Dynamic Countermeasure Fabrics for Post-Spaceflight Orthostatic Intolerance

Aerospace orthostatic intolerance garments (OIG) have historically been pneumatic (e.g., NASA’s antigravity suit), an approach that inhibits mobility and requires connection to an air supply. Elastic compression garments, an alternative technology, are difficult to don/doff and cannot be worn in a noncompressive state, resulting in discomfort and usability challenges. This research evaluates a novel technology—contractile shape memory alloy (SMA) knitted actuators—that can enable low-profile, dynamic compression for an aerospace OIG. To characterize the functional capabilities of SMA knitted actuators, displacement control testing was conducted on 10 actuator samples with a range of geometric design parameters. Inactive (FI) and actuated forces (FA) were observed by repeatedly thermally cycling each sample at 0%, 15%, 30%, and 45% structural strain. Compression capabilities were approximated using medical compression hosiery standards and anthropometric data from a representative aerospace population (ANSUR 2012). Dynamic compression predictions reached 52 mmHg (single layer fabric) and 105 mmHg (double layer fabric) at the ankle. Low, inactive pressures (p, 20 mmHg) demonstrate that compression is controllable and can be dynamically increased upon actuation up to 33 mmHg in a single layer system and up to 67 mmHg in a double layer system. The results highlight the potential of SMA knitted actuators to enable low-profile, dynamic compression garments that can reach medically therapeutic pressures on an aerospace population to counteract OI symptoms. In addition to astronautic applications, this technology demonstrates widespread terrestrial medical and high-performance aircraft applicability.

Orthostatic intolerance↗

Parallel Variable Population Multi-Objective Optimizer (pvpmoo) v1.0

This is a parallel variable population multi-objective optimizer with an adaptive unified differential evolution algorithm or a genetic algorithm. It can also be used for single objective optimization. Some features of this code include: 1) The population size varies from generation to generation to save the total # of objective function evaluations. 2) The population is uniformly distributed to a number of parallel processors for simultaneous objective function evaluation. 3) The objective function evaluation can be attained from an external simulation program with control variables in its input file and objectives calculated from its output files. 4) The optimizer includes an adaptive unified differential evolution algorithm and a real value genetic algorithm. The parameters in the unified differential evolution algorithm can be chosen to attain any mutation schemes in the published literature.

Qiang, Ji↗

Long term microparticle impact fluxes on LDEF determined from optical survey of Interplanetary Dust Experiment (IDE) sensors

Many of the IDE metal-oxide-silicon (MOS) capacitor-discharge impact sensors remained active during the entire Long Duration Exposure Facility (LDEF) mission. An optical survey of impact sites on the active surfaces of these sensors has been extended to include all sensors from the low-flux sides of LDEF (i.e. the west or trailing side, the earth end, and the space end) and 5-7 active sensors from each LDEF's high-flux sides (i.e. the east or leading side, the south side, and the north side). This survey was facilitated by the presence of a relatively large (greater than 50 micron diameter) optical signature associated with each impact site on the active sensor surfaces. Of the approximately 4700 impacts in the optical survey data set, 84% were from particles in the 0.5 to 3 micron size range. An estimate of the total number of hypervelocity impacts on LDEF from particles greater than 0.5 micron diameter yields a value of approximately 7 x 10(exp 6). Impact feature dimensions for several dozen large craters on MOS sensors and germanium witness plates are also presented. Impact fluxes calculated from the IDE survey data closely matched surveys of similar size impacts (greater than or equal to 3 micron diameter craters in Al, or marginal penetrations of a 2.4 micron thick Al foil) by other LDEF investigators. Since the first year IDE data were electronically recorded, the flux data could be divided into three long term time periods: the first year, the entire 5.8 year mission, and the intervening 4.8 years (by difference). The IDE data show that there was an order of magnitude decrease in the long term microparticle impact flux on the trailing side of LDEF, from 1.01 to 0.098 x 10(exp -4) m(exp 2)/s, from the first year in orbit compared to years 2-6. The long term flux on the leading edge showed an increase from 8.6 to 11.2 x 10(exp -4) m(exp -2)/s over this same time period. (Short term flux increases up to 10,000 times the background rate were recorded on the leading side during LDEF's first year in orbit.) The overall east/west ratio was 44, but during LDEF's first year in orbit the ratio was 8.5, and during years 2-6 the ratio was 114. Long term microparticle impact fluxes on the space end decreased from 1.12 to 0.55 x 10(exp -4) m(exp -2)/s from the first year in orbit compared to years 2-6. The earth end showed the opposite trend with an increase from 0.16 to 0.38 x 10(exp -4) m(exp -2)/s. Fluxes on rows 6 and 12 decreased from 6.1 to 3.4 and 6.7 to 3.7 x 10(exp -4) m(exp -2)/s, respectively, over the same time periods. This resulted in space/earth microparticle impact flux ratios of 7.1 during the first year and 1.5 during years 2-6, while the south/north, space/north and space/south ratios remained constant at 1.1, 0.16 and 0.17, respectively, during the entire mission. This information indicates the possible identification of long term changes in discrete microparticle orbital debris component contributions to the total impact flux experienced by LDEF. A dramatic decrease in the debris population capable of striking the trailing side was detected that could possibly be attributed to the hiatus of western launch activity experienced from 1986-1989. A significant increase in the debris population that preferentially struck the leading side was also observed and could possibly be attributed to a single breakup event that occurred in September of 1986. A substantial increase in the microparticle debris population that struck the earth end of LDEF, but not the space end, was also detected and could possibly be the result of a single breakup event at low altitude. These results point to the importance of including discrete orbital debris component contribution changes in flux models in order to achieve accurate predictions of the microparticle environment that a particular spacecraft will experience in earth orbit. The only reliable, verified empirical measurements of these changes are reported in this paper. Further time-resolved in-situ measurements of these debris populations are needed to accurately assess model predictions and mitigation practices.

Simon, C. G.↗

Phonon-mediated temperature dependence of Er 3+ optical transitions in Er 2 O 3

Characterization of the atomic level processes that determine optical transitions in emerging materials is critical to the development of new platforms for classical and quantum networking. Such understanding often emerges from studies of the temperature dependence of the transitions. We report measurements of the temperature dependent Er 3+ photoluminescence in single crystal Er 2 O 3 thin films epitaxially grown on Si(111) focused on transitions that involve the closely spaced Stark-split levels. Radiative intensities are compared to a model that includes relevant Stark-split states, single phonon-assisted excitations, and the well-established level population redistribution due to thermalization. This approach, applied to the individual Stark-split states and employing Er 2 O 3 specific single-phonon-assisted excitations, gives good agreement with experiment. This model allows us to demonstrate the difference in the electron-phonon coupling of the 4 S 3/2 and 2 H 11/2 states of Er 3+ in E 2 O 3 and suggests that the temperature dependence of Er 3+ emission intensity may vary significantly with small shifts in the wavelength (~0.1 nm) of the excitation source.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Single-cell profiling of Arabidopsis leaves to Pseudomonas syringae infection

Plant response to pathogen infection varies within a leaf, yet this heterogeneity is not well resolved. We expose Arabidopsis to Pseudomonas syringae or mock treatment and profile >11,000 individual cells using single-cell RNA sequencing. Integrative analysis of cell populations from both treatments identifies distinct pathogen-responsive cell clusters exhibiting transcriptional responses ranging from immunity to susceptibility. Pseudotime analyses through pathogen infection reveals a continuum of disease progression from an immune to a susceptible state. Confocal imaging of promoter-reporter lines for transcripts enriched in immune cell clusters shows expression surrounding substomatal cavities colonized or in close proximity to bacterial colonies, suggesting that cells within immune clusters represent sites of early pathogen invasion. Susceptibility clusters exhibit more general localization and are highly induced at later stages of infection. Overall, our work shows cellular heterogeneity within an infected leaf and provides insight into plant differential response to infection at a single-cell level.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluating inkjet dispenser/liquid vortex capture-mass spectrometry for single-cell metabolomics in Hep G2 steatosis caused by tamoxifen

Single-cell mass spectrometry (MS) is advancing our understanding of metabolic pathways in heterogeneous cell populations; however, many techniques are slow or require disruptive sample preparations. This study evaluated coupling a modified HP D100 single-cell inkjet dispenser with liquid vortex capture-mass spectrometry (D100/LVC-MS). The D100 is a single-cell inkjet dispenser capable of titrating solutions and isolating single cells via disposable cassettes equipped with microfluidic channels and an impedance sensor. The LVC-MS enables high-throughput capture, lysis, and ionization of analytes for mass spectrometric analysis. The D100/LVC-MS system was characterized through titration and single-cell experiments. Propranolol titration demonstrated linearity across a broad concentration range using the D100/LVC-MS system. Additionally, Hep G2 hepatocarcinoma cells and Chlamydomonas reinhardtii algae were used to showcase the D100’s high-throughput or low-buffer-volume single-cell dispensing strategies. The D100/LVC-MS system’s performance was validated by evaluating tamoxifen-induced steatosis in Hep G2 cells. Tamoxifen, associated with nonalcoholic fatty liver disease in breast cancer patients following long-term use, was tested in Hep G2 cells at 20 µM for 72 h against DMSO-treated controls. High-throughput analysis of 500 cells per condition, completed in 25 min per run, demonstrated the system’s efficiency. The D100/LVC-MS system simultaneously quantified tamoxifen and measured triglycerides and phosphatidylcholines. Triglycerides were upregulated in the tamoxifen-treated cells and the results indicated two distinct cell populations, differing in tamoxifen and phosphatidylcholines levels, suggesting heterogeneity within the treated population. In conclusion, these findings highlight the D100/LVC-MS system as a cost-effective, high-throughput platform for single-cell metabolomics and lipidomics, with significant potential for evaluating metabolic alterations.

Chromatography↗