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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Differential localization patterns of pyruvate kinase isoforms in murine naïve, formative, and primed pluripotent states

Highlights: • PKM1/2 protein abundance is greater in formative mEpiLCs compared to naïve mESCs or primed mEpiSCs. The ratio of PKM1/2 is maintained. • PKM1/2 are localized in both nuclear and cytoplasmic regions relative to GAPDH and OCT4 across the pluripotent continuum. • PKM1 localization is strongly correlated to OCT4 localization, and moderately to GAPDH in formative mEpiLCs. • PKM1/2 localization is moderately correlated to OCT4 and GAPDH localization in naïve mESCs. • PKM1/2 localization are moderately correlated to GAPDH localization in primed mEpiSCs. Mouse embryonic stem cells (mESCs) and mouse epiblast stem cells (mEpiSCs) represent opposite ends of the pluripotency continuum, referred to as naïve and primed pluripotent states, respectively. These divergent pluripotent states differ in several ways, including growth factor requirements, transcription factor expression, DNA methylation patterns, and metabolic profiles. Naïve cells employ both glycolysis and oxidative phosphorylation (OXPHOS), whereas primed cells preferentially utilize aerobic glycolysis, a trait shared with cancer cells referred to as the Warburg Effect. Until recently, metabolism has been regarded as a by-product of cell fate, however, evidence now supports metabolism as being a driver of stem cell state and fate decisions. Pyruvate kinase muscle isoforms (PKM1 and PKM2) are important for generating and maintaining pluripotent stem cells (PSCs) and mediating the Warburg Effect. Both isoforms catalyze the final, rate limiting step of glycolysis, generating adenosine triphosphate and pyruvate, however, the precise role(s) of PKM1/2 in naïve and primed pluripotency is not well understood. The primary objective of this study was to characterize the cellular expression and localization patterns of PKM1 and PKM2 in mESCs, chemically transitioned epiblast-like cells (mEpiLCs) representing formative pluripotency, and mEpiSCs using immunoblotting and confocal microscopy. The results indicate that PKM1 and PKM2 are not only localized to the cytoplasm, but also accumulate in differential subnuclear regions of mESC, mEpiLCs, and mEpiSCs as determined by a quantitative confocal microscopy employing orthogonal projections and airyscan processing. Importantly, we discovered that the subnuclear localization of PKM1/2 changes during the transition from mESCs, mEpiLCs, and mEpiSCs. Finally, we have comprehensively validated the appropriateness and power of the Pearson's correlation coefficient and Manders's overlap coefficient for assessing nuclear and cytoplasmic protein colocalization in PSCs by immunofluorescence confocal microscopy. We propose that nuclear PKM1/2 may assist with distinct pluripotency state maintenance and lineage priming by non-canonical mechanisms. These results advance our understanding of the overall mechanisms controlling naïve, formative, and primed pluripotency.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2020

Confocal Raman Microscopy Investigation of Phospholipid Monolayers Deposited on Nitrile-Modified Surfaces in Porous Silica Particles

Phospholipid bilayers deposited on a variety of surfaces offer models for investigation of the lipid membrane structure and supports for biocompatible sensors. Hybrid-supported phospholipid bilayers (HSLBs) are stable membrane models for these investigations, typically prepared by self-assembly of a lipid monolayer over an n-alkane-modified surface. HSLBs have been prepared on n-alkyl chain-modified silica and used for lipophilicity-based chromatographic separations. The structure of these hybrid bilayers differs from vesicle membranes where the lipid head group spacing is greater due to interdigitation of the lipid acyl chains with the underlying n-alkyl chains bound to the silica surface. This interdigitated structure exhibits a broader melting transition at a higher temperature due to strong interactions between the lipid acyl chains and the immobile n-alkyl chains bound to silica. In the present work, we seek to reduce the interactions between a lipid monolayer and its supporting substrate by self-assembly of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) on porous silica functionalized with nitrile-terminated surface ligands. The frequency of Raman scattering of the surface -C≡N stretching mode at the lipid–nitrile interface is consistent with an n-alkane-like environment and insensitive to lipid head group charge, indicating that the lipid acyl chains are in contact with the surface nitrile groups. The head group area of this lipid monolayer was determined from the within-particle phospholipid concentration and silica specific surface area and found to be 54 ± 2 Å 2 , equivalent to the head group area of a DMPC vesicle bilayer. The structure of these nitrile-supported phospholipid monolayers was characterized below and above their melting transition by confocal Raman microscopy and found to be nearly identical to DMPC vesicle bilayers. Their narrow gel-to-fluid-phase melting transition is equivalent to dispersed DMPC vesicles, indicating that the acyl chain structure on the nitrile support mimics the outer leaflet structure of a vesicle membrane.

36 MATERIALS SCIENCE↗
DOE OSTI2025

3013 Baseline Inner Container Report: Description, Imaging, and Analysis

This report presents consolidated findings from three microscopy studies of baseline Bagless Transfer Containers (BTCs), which are the inner containers used in DOE Standard 3013-compliant plutonium storage packages. The studies were conducted by Los Alamos National Laboratory (LANL), Savannah River National Laboratory (SRNL), and DNV, a subcontractor specializing in corrosion science. The baseline BTCs examined had not been exposed to corrosive environments, serving as control samples for comparison with destructively examined (DE) containers. Additionally, the Background section provides an overview of the BTC design, including manufacturing methods, material properties, and container configurations that may influence corrosion behavior. The primary objective of the LANL and SRNL studies was to establish reference characteristics of BTCs to distinguish manufacturing artifacts from corrosion-induced features observed in DE containers. Both laboratories used advanced microscopy techniques, including Wide Area Microscopy (WAMS), Laser Confocal Microscopy (LCM), Scanning Electron Microscopy (SEM), and Focused Ion Beam (FIB), to reveal that most surface anomalies were shallow and attributable to flow-forming processes, not corrosion. Subsurface impurities, identified as non-metallic inclusions, were also observed. DNV’s investigation focused on characterizing the mechanical and chemical properties of the BTC material to support crack growth modeling. Their findings indicate higher hardness and strength but reduced ductility—consistent with flow-formed 304L stainless steel and a higher-than-expected martensite content, in contrast to manufacturer claims. Ongoing and future studies, including Electron Backscatter Diffraction (EBSD), aim to further understand the microstructural factors influencing crack initiation and propagation. The combined studies provide a crucial baseline for distinguishing corrosion effects in DE containers, contributing to the safe and reliable long-term storage of plutonium-bearing materials.

36 MATERIALS SCIENCE↗
DOE OSTI2022

Electropolishing of a Full-Sized U-10Mo Plate

Electropolishing is used to remove material from the surface of a metal using electric potential and current. U-10Mo fuel is produced from a low enriched uranium plate alloyed with 10% molybdenum (U-10Mo). Electropolishing is being considered in two steps of the process of fabricating this U-10Mo fuel. First, it could be used as a means initially improving the surface finish of the cast plate while removing smut from the casting process from the surface prior to homogenization. Second, it could be used again after homogenization to remove oxidation and continue to improve the surface finish prior to hot rolling. The purpose of this work is to demonstrate the process on full-size ingots and optimize the process. A depleted uranium U-10Mo plate was electropolished three times: the first two times were prior to homogenization and the third after a homogenization step. The first two polishes decreased the overall roughness, as measured by laser confocal microscopy. After each polish, a smoother and shinier blue-gold surface was left on the plate. Upon homogenization, the surface had a more matte appearance and the surface roughness increased back to the pre-homogenization values obtained prior to the initial polish. After electropolishing the homogenized pieces, the changes in surface roughness increased, as measured by laser confocal microscopy. The final polishing step was not long enough to return the surface roughness to pre-homogenization conditions, implying less polishing is needed before the homogenization versus afterwards. Heat generation during the polish was addressed by circulating the electrolyte solution through an external heat exchanger; however, solubility and anodization challenges require further study.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗
DOE OSTI2023

Flash NanoPrecipitation as an Agrochemical Nanocarrier Formulation Platform: Phloem Uptake and Translocation after Foliar Administration

The increasing severity of pathogenic and environmental stressors that negatively affect plant health has led to interest in developing next-generation agrochemical delivery systems capable of precisely transporting active agents to specific sites within plants. In this work, we adapt Flash NanoPrecipitation (FNP), a scalable nanocarrier (NC) formulation technology used in the pharmaceutical industry, to prepare organic core–shell NCs and study their efficacy as foliar or root delivery vehicles. NCs ranging in diameter from 55 to 200 nm, with surface zeta potentials from -40 to +40 mV, and with seven different shell material properties were prepared and studied. Shell materials included synthetic polymers poly(acrylic acid), poly(ethylene glycol), and poly(2-(dimethylamino)ethyl methacrylate), naturally occurring compounds fish gelatin and soybean lecithin, and semisynthetic hydroxypropyl methylcellulose acetate succinate (HPMCAS). NC cores contained a gadolinium tracer for tracking by mass spectrometry, a fluorescent dye for tracking by confocal microscopy, and model hydrophobic compounds (alpha tocopherol acetate and polystyrene) that could be replaced by agrochemical payloads in subsequent applications. After foliar application onto tomato plants with Silwet L-77 surfactant, internalization efficiencies of up to 85% and NC translocation efficiencies of up to 32% were observed. Significant NC trafficking to the stem and roots suggests a high degree of phloem loading for some of these formulations. Results were corroborated by confocal microscopy and synchrotron X-ray fluorescence mapping. NCs stabilized by cellulosic HPMCAS exhibited the highest degree of translocation, followed by formulations with a significant surface charge. The results from this work indicate that biocompatible materials like HPMCAS are promising agrochemical delivery vehicles in an industrially viable pharmaceutical nanoformulation process (FNP) and shed light on the optimal properties of organic NCs for efficient foliar uptake, translocation, and delivery.

36 MATERIALS SCIENCE↗
DOE OSTI2025

Investigating the FLASH Effect in a Rat Brain Organotypic Model With a Novel High-Energy Electron Beam

Ultrahigh dose rate (FLASH) radiation therapy is reported to reduce normal tissue toxicity while maintaining tumor control; however, mechanism(s) remain obscure. To study FLASH mechanisms in brain tissue, we developed a novel experimental platform featuring a specialized high-energy electron linear accelerator, High Intensity Gamma Ray Source (HIGS), paired with an organotypic ex vivo brain metastasis model. We varied interpulse spacing to modulate the mean dose rate (MDR) of our unique 35 MeV electron beam, while maintaining extremely high instantaneous dose rate (IDR). We characterized dosimetry and targeting accuracy of the FLASH beam with film dosimetry. We combined this FLASH beam with an organotypic rat brain slice/breast carcinoma coculture model of brain metastasis to assess effects on normal and neoplastic tissues. Live-cell and bioluminescence imaging demonstrated cancer cell growth effects, whereas normal tissue responses and immune activation were assessed using live-cell imaging, cytokine profiles, and confocal microscopy. Here, we performed comparison experiments with 20 MeV electrons from a Varian clinical linear accelerator (VCLA) using conventional dose rates. The highest IDR of the FLASH beam to date was 20.7 ± 0.6 MGy/s, with maximum MDR of 20.7 MGy/s delivered in 1 pulse of 1 µs duration. Beam targeting was accurate to <1 mm and reproducible. HIGS-FLASH and VCLA dose rates equivalently decreased cancer cell growth. HIGS-FLASH irradiation significantly increased tumor necrosis factor α and fractalkine levels and confocal microscopy revealed distinct changes in microglial morphology slices suggesting microglia activation. Our novel experimental platform produces extremely high dose rates and rapid normal/neoplastic tissue readouts for mechanistic research into the effects of FLASH radiation in the brain. HIGS-FLASH irradiation induces comparable cancer cell growth inhibition but differential effects on cytokines and microglial morphology, suggesting that acute innate immune responses may be involved in FLASH normal tissue effects in the brain.

Kay, Tyler V. [Duke University, Durham, NC (United↗
DOE OSTI2021

DeepLearnMOR: a deep-learning framework for fluorescence image-based classification of organelle morphology

Abstract The proper biogenesis, morphogenesis, and dynamics of subcellular organelles are essential to their metabolic functions. Conventional techniques for identifying, classifying, and quantifying abnormalities in organelle morphology are largely manual and time-consuming, and require specific expertise. Deep learning has the potential to revolutionize image-based screens by greatly improving their scope, speed, and efficiency. Here, we used transfer learning and a convolutional neural network (CNN) to analyze over 47,000 confocal microscopy images from Arabidopsis wild-type and mutant plants with abnormal division of one of three essential energy organelles: chloroplasts, mitochondria, or peroxisomes. We have built a deep-learning framework, DeepLearnMOR (Deep Learning of the Morphology of Organelles), which can rapidly classify image categories and identify abnormalities in organelle morphology with over 97% accuracy. Feature visualization analysis identified important features used by the CNN to predict morphological abnormalities, and visual clues helped to better understand the decision-making process, thereby validating the reliability and interpretability of the neural network. This framework establishes a foundation for future larger-scale research with broader scopes and greater data set diversity and heterogeneity.

Plant Sciences↗
DOE OSTI2023

Plasmonic nanorod probes’ journey inside plant cells for in vivo SERS sensing and multimodal imaging

Nanoparticle-based platforms are gaining strong interest in plant biology and bioenergy research to monitor and control biological processes in whole plants. However, in vivo monitoring of biomolecules using nanoparticles inside plant cells remains challenging due to the impenetrability of the plant cell wall to nanoparticles beyond the exclusion limits (5-20 nm). To overcome this physical barrier, we have designed unique bimetallic silver-coated gold nanorods (AuNR@Ag) capable of entering plant cells, while conserving key plasmonic properties in the near-infrared (NIR). To demonstrate cellular internalization and tracking of the nanorods inside plant tissue, we used a comprehensive multimodal imaging approach that included transmission electron microscopy (TEM), confocal fluorescence microscopy, two-photon luminescence (TPL), X-ray fluorescence microscopy (XRF), and photoacoustics imaging (PAI). We successfully acquired SERS signals of nanorods in vivo inside plant cells of tobacco leaves. On the same leaf samples, we applied orthogonal imaging methods, TPL and PAI techniques for in vivo imaging of the nanorods. This study first demonstrates the intracellular internalization of AuNR@Ag inside whole plant systems for in vivo SERS analysis in tobacco cells. This work demonstrates the potential of this nanoplatform as a new nanotool for intracellular in vivo biosensing for plant biology.

Cupil-Garcia, Vanessa↗
DOE OSTI2021

Imaging of surface acoustic waves on GaAs using 2D confocal Raman microscopy and atomic force microscopy

Surface acoustic wave devices have been fabricated on a GaAs (100) substrate to demonstrate the capability of 2D Raman microscopy as an imaging technique for acoustic waves on the surface of a piezoelectric substrate. Surface acoustic waves are generated using a two-port interdigitated transducer platform, which is modified to produce surface standing waves. We have derived an analytical model to relate Raman peak broadening to the near-surface strain field of the GaAs surface produced by the surface acoustic waves. Additionally, atomic force microscopy is used to confirm the presence of a standing acoustic wave, resolving a total vertical displacement of 3 nm at the antinode of the standing wave. Stress calculations are performed for both imaging techniques and are in good agreement, demonstrating the potential of this Raman analysis.

47 OTHER INSTRUMENTATION↗
DOE OSTI2020

Bifacial Multilayer Graphene Float Transfer

A method for graphene transfer which is referred to as “bifacial transfer” that allows transfer of multilayer chemical vapor deposition (CVD) graphene from both sides of a native metal substrate, such as an as-received nickel catalyst, is presented. In traditional transfer methods, the graphene on the “non-preferred” side, that is, the bottom of the substrate, is removed with oxygen plasma before removal of the metal catalyst in etchant solution. Although this treatment prevents undesired aggregation of the graphene films, it fails to utilize both sides of CVD-grown graphene. The bifacial transfer method reduces the cost of multilayer graphene by allowing the transfer of graphene from both sides of the substrate. The quality of graphene transferred from both sides onto target glass and polymer substrates is compared. The results of optical microscopy, confocal Raman spectroscopy, atomic force microscopy, and electronic transport measurements suggest that the quality of the multilayer graphene on the “non-preferred” side does not differ significantly from that of the “preferred” side. Overall, this method will allow more efficient and cost-effective use of graphene by doubling the usable graphene per area of growth substrate, and by eliminating the need for intermediate sacrificial transfer substrates such as poly(methyl methacrylate).

77 NANOSCIENCE AND NANOTECHNOLOGY↗
DOE OSTI2026

Coupling Microstructural Evolution Simulations to Material Property Degradation Predictions for Plasma-Facing Materials

Reliable material performance is required for plasma-facing material (PFM) candidates. Previous research has shown that plasma and neutron radiation exposure induces microstructural changes in PFMs; changes in thermal and electrical conductivities and in material hardening and embrittlement were also observed after neutron irradiation. These material property changes will negatively impact the performance of the PFMs in a fusion reactor. Despite the well-known connection between material microstructure, properties, and performance, there is a need for validated modeling capabilities connecting PFM property degradation with microstructural evolution under fusion-relevant conditions. We are developing a simulation capability to couple plasma-induced microstructural evolution to material property degradation. Our approach relies on deliberate mapping between individual simulation models and experimental characterization for validation. The open-source Multiphysics Object-Oriented Simulation Environment (MOOSE) software was used for this simulation capability development. A MOOSE phase-field model was coupled with the cluster dynamics code, Xolotl, to predict microstructural evolution. Microstructure characterization techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), and laser scanning confocal microscopy (LSCM) are used to validate these microstructural evolution simulations. Calculation of thermal and electrical conductivities with first principles simulations was performed for bulk material and for grain boundaries; these results are used within MOOSE models to calculate effective thermal and electrical conductivities as a function of grain characteristics. Thermoreflectance and four-probe techniques were employed to measure the thermal and electrical conductivities, respectively. A MOOSE crystal plasticity model was adapted to predict microstructure-sensitive deformation behavior, and X-ray diffraction (XRD) was used to collect bulk dislocation density data for validation. After individual simulation validation, these models are coupled to predict material property changes resulting from plasma exposure. We focused here on an experimental design to emphasize the separate effects of moderate thermal loads and plasma exposure using tungsten. Annealing of tungsten was performed under a protective environment for temperatures ranging from 500 C to 1500 C. The plasma exposure was completed in the Tritium Plasma Experiment at Idaho National Laboratory under a deuterium flux of 1e22 D/m^2-s. This incremental approach is employed to build confidence in the modeling capability: separate-effects tests ensure that the models capture key mechanisms from single environmental conditions before predicting PFM property degradation under combined loads. We will show our early results from coupling these simulation models to predict PFM property changes from microstructural evolution. Comparisons of the simulation results with preliminary validation data will be discussed.

36 - MATERIALS SCIENCE↗
DOE OSTI2026

Connect microstructure evolution to property degradation with validated simulation

Reliable material performance is required for plasma-facing material (PFM) candidates. Previous research has shown that plasma and neutron radiation exposure induces microstructural changes in PFMs; changes in thermal and electrical conductivities and in material hardening and embrittlement were also observed after neutron irradiation. These material property changes will negatively impact the performance of the PFMs in a fusion reactor. Despite the well-known connection between material microstructure, properties, and performance, there is a need for validated modeling capabilities connecting PFM property degradation with microstructural evolution under fusion-relevant conditions. We are developing a simulation capability to couple plasma-induced microstructural evolution to material property degradation. Our approach relies on deliberate mapping between individual simulation models and experimental characterization for validation. The open-source Multiphysics Object-Oriented Simulation Environment (MOOSE) software was used for this simulation capability development. A MOOSE phase-field model was coupled with the cluster dynamics code, Xolotl, to predict microstructural evolution. Microstructure characterization techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), and laser scanning confocal microscopy (LSCM) are used to validate these microstructural evolution simulations. Calculation of thermal and electrical conductivities with first principles simulations was performed for bulk material and for grain boundaries; these results are used within MOOSE models to calculate effective thermal and electrical conductivities as a function of grain characteristics. Thermoreflectance and four-probe techniques were employed to measure the thermal and electrical conductivities, respectively. A MOOSE crystal plasticity model was adapted to predict microstructure-sensitive deformation behavior, and X-ray diffraction (XRD) was used to collect bulk dislocation density data for validation. After individual simulation validation, these models are coupled to predict material property changes resulting from plasma exposure. We focused here on an experimental design to emphasize the separate effects of moderate thermal loads and plasma exposure using tungsten. Annealing of tungsten was performed under a protective environment for temperatures ranging from 500$^o$C to 1500$^o$C. The plasma exposure was completed in the Tritium Plasma Experiment at Idaho National Laboratory under a deuterium flux of 1e22 $\frac{D}{m^2s}$. This incremental approach is employed to build confidence in the modeling capability: separate-effects tests ensure that the models capture key mechanisms from single environmental conditions before predicting PFM property degradation under combined loads. We will show our early results from coupling these simulation models to predict PFM property changes from microstructural evolution. Comparisons of the simulation results with preliminary validation data will be discussed.

36 - MATERIALS SCIENCE↗
DOE OSTI2020

Antibody Targeted Metal–Organic Frameworks for Bioimaging Applications

Here, we report on the availability and chemical utility of primary amines within metal–organic frameworks (MOFs) for cell targeting. Primary amine groups represent one of the most versatile chemical moieties for conjugation to biologically relevant molecules, including antibodies and enzymes. Specifically, we used two different chemical conjugations schemes, utilizing the amino functionality on the organic linker: first, carbodiimide chemistry was used to link the primary amine to available carboxyl groups on the protein neutravidin; second, sulfhydryl cross-linking chemistry was used via Traut’s reagent scheme. Importantly, this is the first report that documents this methodology implemented with MOF systems. Finally, the ability of the EpCAM antibody targeted MOFs to bind to a human epithelial cell line (A549), a common target for imaging studies, was confirmed with confocal microscopy.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Biofouling of filtration membranes in wastewater reuse: In situ visualization with confocal laser scanning microscopy

The filtration membranes utilized in advanced treatment of municipal wastewater are prone to biological fouling. Permeability loss and water recovery limitations due to fouling necessitate additional pretreatment and membrane cleaning, which raise the cost of water reuse. Better fouling mitigation calls for a deeper understanding of biofilm behavior on water reuse membranes. This study uses confocal microscopy to monitor biofouling under realistic conditions. To simulate water reuse, a miniature flow cell with microfiltration and ultrafiltration membranes was operated under typical filtration conditions using secondary effluent from California's East Bay Municipal Utility District. The growing biofilms were stained for extracellular polymeric substances and live and dead cells, and they were monitored in situ under filtration conditions with a confocal laser scanning microscope to produce 2-D and 3-D images. In situ image series revealed biofilm growth and removal processes including internal and external fouling, extracellular polymeric substance production, wrinkling, delamination, and film re-deposition. Additionally, changes in biofilm morphology between in situ and ex situ images highlighted limitations of ex situ imaging. Here, these results provide insight into the physical and biological mechanisms of biofouling in hopes of informing the development of improved techniques for biofouling mitigation.

ultrafiltration↗
DOE OSTI2021

Control of monodomain polymer-stabilized cuboidal nanocrystals of chiral nematics by confinement.

Liquid crystals are important components of optical technologies. Cuboidal crystals consisting of chiral liquid crystals-the so-called blue phases (BPs), are of particular interest due to their crystalline structures and fast response times, but it is critical that control be gained over their phase behavior as well as the underlying dislocations and grain boundaries that arise in such systems. Blue phases exhibit cubic crystalline symmetries with lattice parameters in the 100 nm range and a network of disclination lines that can be polymerized to widen the range of temperatures over which they occur. Here, we introduce the concept of strain-controlled polymerization of BPs under confinement, which enables formation of strain-correlated stabilized morphologies that, under some circumstances, can adopt perfect single-crystal monodomain structures and undergo reversible crystal-to-crystal transformations, even if their disclination lines are polymerized. We have used super-resolution laser confocal microscopy to reveal the periodic structure and the lattice planes of the strain and polymerization stabilized BPs in 3D real space. Our experimental observations are supported and interpreted by relying on theory and computational simulations in terms of a free energy functional for a tensorial order parameter. Simulations are used to determine the orientation of the lattice planes unambiguously. The findings presented here offer opportunities for engineering optical devices based on single-crystal, polymer-stabilized BPs whose inherent liquid nature, fast dynamics, and long-range crystalline order can be fully exploited.

confinement↗
DOE OSTI2024

Characterization of Incremental Markings in the Sagittal Otolith of the Pacific Sardine ( Sardinops sagax ) Using Different Imaging Modalities

Teleost fish possess calcium carbonate otoliths located in separate chambers (utriculus, sacculus, and lagena) of their membranous labyrinth. This study analyzed the surface topography of the sagittal otolith of the Pacific sardine (Sardinops sagax) and the daily and annual increments in these otoliths. The otolith surface, characterized by laser scanning confocal microscopy for the first time, consisted of a system of prominent ridges and valleys (grooves), but it is unclear whether these structures are functional or represent time-resolving markings reflecting growth periodicity. Within the first-year volume, daily increments, each consisting of an incremental (more mineralized) and a discontinuous (less mineralized) zone, were resolved by optical microscopy and backscattered electron (BSE) imaging in the scanning electron microscope (SEM). Daily growth increments could, however, not be resolved in volumes formed after the first year, presumably because otolith growth markedly slows down and spacing of incremental markings narrows in older fish. Throughout otolith growth, the crystalline network continues across the discontinuous zones. Fluorochrome labeling provides additional information on growth after the first year. Compared with optical and BSE imaging, synchrotron microComputed Tomography of intact otoliths (with 0.69 µm volume elements) was less able to resolve daily increments; X-ray phase contrast reconstructions provided more detail than reconstructions with absorption contrast. Future research directions are proposed.

59 BASIC BIOLOGICAL SCIENCES↗