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At least 145 records · Page 8

Technology

In this session, Session WP3, the discussion focuses on the following topics: Monitoring Physiological Variables With Membrane Probes; Real Time Confocal Laser Scanning Microscopy, Potential Applications in Space Medicine and Cell Biology; Optimum Versus Universal Planetary and Interplanetary Habitats; Application of Remote Sensing and Geographic Information System Technologies to the Prevention of Diarrheal Diseases in Nigeria; A Small G Loading Human Centrifuge for Space Station ERA; Use of the Bicycle Ergometer on the International Space Station and Its Influence On The Microgravity Environment; Munich Space Chair (MSC) - A Next Generation Body Restraint System for Astronauts; and Thermoelectric Human-Body Cooling Units Used By NASA Space Shuttle Astronauts.

Source record↗

L-citrulline immunostaining identifies nitric oxide production sites within neurons

The cellular and subcellular localization of L-citrulline was analyzed in the adult rat brain and compared with that of traditional markers for the presence of nitric oxide synthase. Light, transmission electron, and confocal laser scanning microscopy were used to study tissue sections processed for immunocytochemistry employing a monoclonal antibody against L-citrulline or polyclonal anti-neuronal nitric oxide synthase sera, and double immunofluorescence to detect neuronal nitric oxide synthase and L-citrulline co-localization. The results demonstrate that the same CNS regions and cell types are labeled by neuronal nitric oxide synthase polyclonal antisera and L-citrulline monoclonal antibodies, using both immunocytochemistry and immunofluorescence. Short-term pretreatment with a nitric oxide synthase inhibitor reduces L-citrulline immunostaining, but does not affect neuronal nitric oxide synthase immunoreactivity. In the vestibular brainstem, double immunofluorescence studies show that many, but not all, neuronal nitric oxide synthase-positive cells co-express L-citrulline, and that local intracellular patches of intense L-citrulline accumulation are present in some neurons. Conversely, all L-citrulline-labeled neurons co-express neuronal nitric oxide synthase. Cells expressing neuronal nitric oxide synthase alone are interpreted as neurons with the potential to produce nitric oxide under other stimulus conditions, and the subcellular foci of enhanced L-citrulline staining are viewed as intracellular sites of nitric oxide production. This interpretation is supported by ultrastructural observations of subcellular foci with enhanced L-citrulline and/or neuronal nitric oxide synthase staining that are located primarily at postsynaptic densities and portions of the endoplasmic reticulum. We conclude that nitric oxide is produced and released at focal sites within neurons that are identifiable using L-citrulline as a marker. Copyright 2002 IBRO.

NASA Discipline Neuroscience↗

Time course changes in [Ca2+]i, force, and protein content in hindlimb-suspended mouse soleus muscles

BACKGROUND: Exposure to reduced gravitational forces during spaceflight is associated with significant reductions in skeletal muscle mass and strength. The purpose of this study was to test the hypothesis that increases in resting cytosolic free calcium concentration ([Ca2+]i) would precede reductions in protein content and maximal isometric tetanic force (Po) in mouse soleus muscle after initiation of hindlimb suspension. METHODS: Female ICR mice (n = 42) were hindlimb suspended for 1, 2, 3, 5, or 7 d; weight-matched mice were used as controls. Following the hindlimb suspension, the left soleus muscle was used to determine Po in vitro and the right soleus muscle was used to determine protein content and [Ca2+]i via confocal laser scanning microscopy. RESULTS: Compared with controls, [Ca2+]i was elevated by 38% at 2 d, and 117% at 7 d. Compared with controls, soleus muscle total and myofibrillar protein contents were reduced 27-29% and 30-34%, respectively, at 5-7 d after initiation of hindlimb suspension. Compared with controls, soleus muscle Po was decreased by 24% at 3 d, and 38% at 7 d. CONCLUSION: It appears that resting cytosolic Ca2+ homeostasis is disturbed soon after the initiation of hindlimb suspension, and these elevations in [Ca2+]i may play a role in initiating soleus muscle atrophy.

Non-NASA Center↗

Hair cell recovery in mitotically blocked cultures of the bullfrog saccule

Hair cells in many nonmammalian vertebrates are regenerated by the mitotic division of supporting cell progenitors and the differentiation of the resulting progeny into new hair cells and supporting cells. Recent studies have shown that nonmitotic hair cell recovery after aminoglycoside-induced damage can also occur in the vestibular organs. Using hair cell and supporting cell immunocytochemical markers, we have used confocal and electron microscopy to examine the fate of damaged hair cells and the origin of immature hair cells after gentamicin treatment in mitotically blocked cultures of the bullfrog saccule. Extruding and fragmenting hair cells, which undergo apoptotic cell death, are replaced by scar formations. After losing their bundles, sublethally damaged hair cells remain in the sensory epithelium for prolonged periods, acquiring supporting cell-like morphology and immunoreactivity. These modes of damage appear to be mutually exclusive, implying that sublethally damaged hair cells repair their bundles. Transitional cells, coexpressing hair cell and supporting cell markers, are seen near scar formations created by the expansion of neighboring supporting cells. Most of these cells have morphology and immunoreactivity similar to that of sublethally damaged hair cells. Ultrastructural analysis also reveals that most immature hair cells had autophagic vacuoles, implying that they originated from damaged hair cells rather than supporting cells. Some transitional cells are supporting cells participating in scar formations. Supporting cells also decrease in number during hair cell recovery, supporting the conclusion that some supporting cells undergo phenotypic conversion into hair cells without an intervening mitotic event.

Non-NASA Center↗

Intracellular Ca2+ transients in mouse soleus muscle after hindlimb unloading and reloading

The objective of this study was to determine whether altered intracellular Ca(2+) handling contributes to the specific force loss in the soleus muscle after unloading and/or subsequent reloading of mouse hindlimbs. Three groups of female ICR mice were studied: 1) unloaded mice (n = 11) that were hindlimb suspended for 14 days, 2) reloaded mice (n = 10) that were returned to their cages for 1 day after 14 days of hindlimb suspension, and 3) control mice (n = 10) that had normal cage activity. Maximum isometric tetanic force (P(o)) was determined in the soleus muscle from the left hindlimb, and resting free cytosolic Ca(2+) concentration ([Ca(2+)](i)), tetanic [Ca(2+)](i), and 4-chloro-m-cresol-induced [Ca(2+)](i) were measured in the contralateral soleus muscle by confocal laser scanning microscopy. Unloading and reloading increased resting [Ca(2+)](i) above control by 36% and 24%, respectively. Although unloading reduced P(o) and specific force by 58% and 24%, respectively, compared with control mice, there was no difference in tetanic [Ca(2+)](i). P(o), specific force, and tetanic [Ca(2+)](i) were reduced by 58%, 23%, and 23%, respectively, in the reloaded animals compared with control mice; however, tetanic [Ca(2+)](i) was not different between unloaded and reloaded mice. These data indicate that although hindlimb suspension results in disturbed intracellular Ca(2+) homeostasis, changes in tetanic [Ca(2+)](i) do not contribute to force deficits. Compared with unloading, 24 h of physiological reloading in the mouse do not result in further changes in maximal strength or tetanic [Ca(2+)](i).

NASA Program Biomedical Research and Countermeasur↗

Synthesis, Characterization, Topographical Modification, and Surface Properties of Copoly(Imide Siloxane)s

Novel copoly(imide siloxane)s were synthesized from commercially available aminopropyl terminated siloxane oligomers, aromatic dianhydrides, and diamines. This synthetic approach produced copolymers with well-defined siloxane blocks linked with imide units in a random fashion. The copoly(amide acid)s were characterized by solution viscosity and subsequently used to cast thin films followed by thermal imidization in an inert atmosphere. Thin films were characterized using contact angle goniometry, attenuated total reflection Fourier transform infrared spectroscopy, confocal and optical microscopy, and tensile testing. Adhesion of micronsized particles was determined quantitatively using a sonication device. The polydimethylsiloxane (PDMS) moieties lowered the copolymer surface energy due to migration of siloxane moieties to the film s surface, resulting in a notable reduction in particle adhesion. A further reduction in particle adhesion was achieved by introducing topographical features on a scale of several to tens of microns by a laser ablation technique.

Wohl, Christopher J.↗

Achieving Improved Reliability with Failure Analysis

Reliability is the ability of a product to properly function, within specified performance limits, for a specified period of time, under the life cycle application conditions. Failure analysis is a vital tool in the effort to ensure reliability of electronic products and systems throughout their product lifecycle. Today, organizations involved in activities within the electronics supply chain are facing new challenges, not just from complex assembly styles, harsher lifecycle environments, and sophisticated supply chains, but also from customers who are demanding a quicker turn-around. Unfortunately, root cause failure analysis is often performed incompletely, leading to a poor understanding of failure mechanisms and causes and, customer dissatisfaction due to recurring failures. The PDC (Professional Development Course) starts with an introduction to reliability concepts, physics of failure and an overview of failure mechanisms that affect PCBs (Printed Circuit Boards), PCBAs (Printed Circuit Board Assembly) and components. The PDC then dives into root cause hypothesizing techniques (Pareto, FMEA (Failure Modes and Effects Analysis), fishbone (Cause-And-Effect Diagram), FTA (Fault Tree Analysis)), non-destructive and destructive analysis and, materials characterization will be discussed. Numerous failure analysis case studies will be used to illustrate the techniques and analysis principles to arrive at the root cause(s) of field failures on printed circuit boards, active components, and assemblies. What Attendees will Learn: Topics include: Overview of Reliability Concepts Failure mechanisms of electronic products Root cause analysis Failure analysis techniques -Non-destructive techniques (optical, CSAM (Confocal Scanning Electron Microscopy) etc.) -Destructive analysis (DPA (Destructive Physical Analysis), Decap (Decapsulation), FIB (Focused Ion Beam) etc.) -Materials characterization (XRF (X-Ray Fluorescence) , EDS (Error Detection Sequential), TMA/DSC (Thermal Mechanical Analysis/Differential Scanning Calorimetry) etc.)

PCB quality↗

The Light Microscopy Module: An On-Orbit Multi-User Microscope Facility

The Light Microscopy Module (LMM) is planned as a remotely controllable on-orbit microscope subrack facility, allowing flexible scheduling and operation of fluids and biology experiments within the Fluids and Combustion Facility (FCF) Fluids Integrated Rack (FIR) on the International Space Station (ISS). The LMM will be the first integrated payload with the FIR to conduct four fluid physics experiments. A description of the LMM diagnostic capabilities, including video microscopy, interferometry, laser tweezers, confocal, and spectrophotometry, will be provided.

Motil, Susan M.↗

Gravity Sensors and the Role of 3-D Visualization and Simulation in Biomedical Research

Ross 3.0 software was developed in the Biocomputation Center for semi-automated reconstruction of objects from serial thin sections. Data are captured directly from a transmission electron microscope via a video camera to a graphics workstation where the sections are mosaicked and contours are traced, registered and displayed by semi-automated methods. For the first time, macular type II cells are described completely for their innervation patterns. The purposes are to learn more about the fundamental circuitry of the macula and to demonstrate whether the terminals are altered morphologically by space flight. Current examples, from the medial part of the macula, are from maculas collected in-flight on the Space Life Sciences-2 mission, 4.5 hrs post-flight, and from a ground control. Results show that the typical type 11 cell receives processes from up to six nearby calyces or afferents. Nearly all the processes are elongated; some have bouton-like swellings and numerous vesicles. Multiple (2 to 4) processes from a single calyx to a type II cell are common, and approx. 1/3 of the processes innervate 2 type II cells of a neighboring cluster of 3 cells. About 2% of type II cells resemble type I cells morphologically and are surrounded by demicalyces. Differences in size or shape of the terminals under flight conditions could not be determined because the sample size is still too small; but it is clear that reconstruction methods provide insights into macular circuitry not obtainable by other techniques. The results demonstrate a morphological basis for interactions between adjacent receptive fields, through feedback-feedforward connections, during preprocessing of linear acceleratory information by the maculas. While the methods are currently being tested using vestibular maculas as the model system, it is clear that the technology is applicable to any tissue that can be physically or optically sectioned. ROSS software has already been implemented for reconstructing objects from tissues studied by confocal and by transmitted light microscopy, and research into magnetic resonance imaging-computational tomography combined visualization are underway.

Ross, Muriel D.↗

Physiological Maturation of Regenerating Hair Cells

The bullfrog saccule, a sensor of gravity and substrate-borne vibration, is a model system for hair cell transduction. Saccular hair cells also increase in number throughout adult life and rapidly recover after hair cell damage, making this organ an ideal system for studying hair cell development, repair, and regeneration. We have used of hair cell and supporting cell immunocytochemical markers to identify damaged hair cells and hair cell precursors in organotypic cultures of the bullfrog saccule. We then used an innovative combination of confocal, electron, and time-lapse microscopy to study the fate of damaged hair cells and the origin of new hair cells after gentamicin ototoxicity in normal and mitotically blocked saccular cultures. These studies have shown that gentamicin ototoxicity produces both lethal and sublethal hair cell damage. They have also shown that hair cell recovery in this organ takes place by both the repair of sublethally damaged hair cells and by the replacement of lost hair cells by mitotic regeneration. In parallel studies, we have used biophysical and molecular biological techniques to study the differentiation and innervation of developing, repairing, and regenerating hair cells. More specifically, we have used RT-PCR to obtain the bullfrog homologues of L-type voltage- gated calcium (L-VGCC) and large-conductance Ca(2+)-activated potassium (BK) channel genes. We have then obtained probes for these genes and, using in situ hybridization, begun to examine their expression in the bullfrog saccule and amphibian papilla. We have also used fluorescent-labeled channel toxins and channel toxin derivatives to determine the time of appearance of L-type voltage-gated calcium (L-VGCC) and Ca(2+)-activated potassium (BK) channels and to study dynamic changes in the number, distribution, and co-localization of these proteins in developing, repairing, and regenerating hair cells. Using time-lapse microscopy, we are also studying the dynamic relationship between ion channel clustering and synaptic formation in hair cells and afferent neurons. In future studies, we will determine when hair cell precursors acquire electrical tuning, and, using whole-cell patch-clamp techniques, identify and characterize their L-VGCC and BK currents. We will also use biophysical techniques to determine the number of L-VGCC and BK channels and the size and gating kinetics of their underlying L-VGCC and BK conductances, correlating these variables with the amplitude and frequency of membrane oscillations produced by intracellular current steps. We expect these studies to determine how hair cells regulate ion channel expression to achieve specific physiological responses.

Baird, Richard A.↗

Identification of a calmodulin-regulated Ca2+-ATPase in the endoplasmic reticulum

A unique subfamily of calmodulin-dependent Ca2+-ATPases was recently identified in plants. In contrast to the most closely related pumps in animals, plasma membrane-type Ca2+-ATPases, members of this new subfamily are distinguished by a calmodulin-regulated autoinhibitor located at the N-terminal instead of a C-terminal end. In addition, at least some isoforms appear to reside in non-plasma membrane locations. To begin delineating their functions, we investigated the subcellular localization of isoform ACA2p (Arabidopsis Ca2+-ATPase, isoform 2 protein) in Arabidopsis. Here we provide evidence that ACA2p resides in the endoplasmic reticulum (ER). In buoyant density sucrose gradients performed with and without Mg2+, ACA2p cofractionated with an ER membrane marker and a typical "ER-type" Ca2+-ATPase, ACA3p/ECA1p. To visualize its subcellular localization, ACA2p was tagged with a green fluorescence protein at its C terminus (ACA2-GFPp) and expressed in transgenic Arabidopsis. We collected fluorescence images from live root cells using confocal and computational optical-sectioning microscopy. ACA2-GFPp appeared as a fluorescent reticulum, consistent with an ER location. In addition, we observed strong fluorescence around the nuclei of mature epidermal cells, which is consistent with the hypothesis that ACA2p may also function in the nuclear envelope. An ER location makes ACA2p distinct from all other calmodulin-regulated pumps identified in plants or animals.

NASA Discipline Plant Biology↗

ROADRUNNER uranium nitride MiniFuel: Experimental design, fabrication and pre-irradiation baseline characterization for accelerated burnup testing

Uranium nitride (UN) is a promising fuel candidate for advanced reactor systems owing to its high uranium density and thermal conductivity; however, its qualification remains constrained by the scarcity of well-controlled irradiation performance data. Here, to address this limitation, the ROADRUNNER (Research On ADvancing the peRformance of UraNium Nitrides in Extreme enviRonments) campaign employs the MiniFuel platform in the High Flux Isotope Reactor (HFIR) to enable accelerated burnup irradiation testing under tightly controlled and largely isothermal conditions. This paper presents the experimental design, fuel fabrication, and pre-irradiation baseline characterization of the ROADRUNNER UN MiniFuel campaign. Thirty-six UN minidisc specimens were fabricated with systematically varied as-fabricated density (86–96% of theoretical density), carbon impurity content (961–5240 ppm), oxygen content (≤ ∼2000 ppm), and grain size (2.5–24 μm). The irradiation matrix spans nominal fuel temperatures of 873 K, 1173 K, and 1473 K and target burnups of 3.75%, 6.0%, and 7.5% fissions per initial metal atom (FIMA). Neutronic and thermal analyses were performed to define specimen-specific burnup accumulation and temperature histories, establishing the boundary conditions for subsequent in-pile behavior. Comprehensive pre-irradiation characterization—including dimensional metrology, density verification, impurity analysis, X-ray diffraction, Raman spectroscopy, scanning electron microscopy, X-ray computed tomography, and confocal profilometry—provides a detailed baseline for post-irradiation examination. Pre-irradiation data were further used to generate predictive estimates of fission gas release and swelling using existing empirical correlations. This quantitative comparison reveals substantial inter-model divergence at intermediate and elevated temperatures that exceeds propagated input uncertainties, highlighting structural gaps in the historical irradiation database. The ROADRUNNER irradiation campaign is currently underway in HFIR, with initial firs cycle completed in late 2025 and remaining targets scheduled through 2027. The experimental design and baseline dataset presented here establish the framework needed to interpret forthcoming post-irradiation measurements and to provide discriminating data for the validation and refinement of physics-based UN fuel performance models.

Lopes, Denise Adorno [Oak Ridge National Laborator↗

Two-Photon Fluorescence Microscopy for Biomedical Research

This viewgraph presentation gives an overview of two-photon microscopy as it applies to biomedical research. The topics include: 1) Overview; 2) Background; 3) Principles of Operation; 4) Advantages Over Confocal; 5) Modes of Operation; and 6) Applications.

Fischer, David↗

Dense, continuous membrane labeling and expansion microscopy visualization of ultrastructure in tissues

Abstract Lipid membranes are key to the nanoscale compartmentalization of biological systems, but fluorescent visualization of them in intact tissues, with nanoscale precision, is challenging to do with high labeling density. Here, we report ultrastructural membrane expansion microscopy (umExM), which combines an innovative membrane label and optimized expansion microscopy protocol, to support dense labeling of membranes in tissues for nanoscale visualization. We validate the high signal-to-background ratio, and uniformity and continuity, of umExM membrane labeling in brain slices, which supports the imaging of membranes and proteins at a resolution of ~60 nm on a confocal microscope. We demonstrate the utility of umExM for the segmentation and tracing of neuronal processes, such as axons, in mouse brain tissue. Combining umExM with optical fluctuation imaging, or iterating the expansion process, yields ~35 nm resolution imaging, pointing towards the potential for electron microscopy resolution visualization of brain membranes on ordinary light microscopes.

Science & Technology - Other Topics↗

Light Microscopy Module: International Space Station Premier Automated Microscope

The Light Microscopy Module (LMM) was launched to the International Space Station (ISS) in 2009 and began hardware operations in 2010. It continues to support Physical and Biological scientific research on ISS. During 2016, if all goes as planned, three experiments will be completed: [1] Advanced Colloids Experiments with Heated base-2 (ACE-H2) and [2] Advanced Colloids Experiments with Temperature control (ACE-T1). Preliminary results, along with an overview of present and future LMM capabilities will be presented; this includes details on the planned data imaging processing and storage system, along with the confocal upgrade to the core microscope. [1] a consortium of universities from the State of Kentucky working through the Experimental Program to Stimulate Competitive Research (EPSCoR): Stuart Williams, Gerold Willing, Hemali Rathnayake, et al. and [2] from Chungnam National University, Daejeon, S. Korea: Chang-Soo Lee, et al.

Microgravity Biology Research↗

Using the Light Microscopy Module (LMM) on the International Space Station (ISS), The Advanced Colloids Experiment (ACE) and MacroMolecular Biophysics (MMB)

The Light Microscopy Module (LMM) was launched to the International Space Station (ISS) in 2009 and began science operations in 2010. It continues to support Physical and Biological scientific research on ISS. During 2016, if all goes as planned, three experiments will be completed: [1] Advanced Colloids Experiments with Heated base-2 (ACE-H2) and [2] Advanced Colloids Experiments with Temperature control (ACE-T1). Preliminary results, along with an overview of present and future LMM capabilities will be presented; this includes details on the planned data imaging processing and storage system, along with the confocal upgrade to the core microscope. [1] a consortium of universities from the State of Kentucky working through the Experimental Program to Stimulate Competitive Research (EPSCoR): Stuart Williams, Gerold Willing, Hemali Rathnayake, et al. and [2] from Chungnam National University, Daejeon, S. Korea: Chang-Soo Lee, et al.

Microgravity Biology Research↗

Quantifying Twist Angles in Cuprate Heterostructures with Anisotropic Raman Signatures

Artificially engineered twisted van der Waals (vdW) heterostructures have unlocked new pathways for exploring emergent quantum phenomena and strongly correlated electronic states. Many of these phenomena are highly sensitive to the twist angle, which can be deliberately tuned to tailor the interlayer interactions. This makes the twist angle a critical tunable parameter, emphasizing the need for precise control and accurate characterization during device fabrication. In particular, twisted cuprate heterostructures based on Bi 2 Sr 2 CaCu 2 O 8 + x (BSCCO) have demonstrated angle-dependent superconducting properties, positioning the twist angle as a key tunable parameter. However, the twisted interface is highly unstable under ambient conditions and vulnerable to damage from conventional characterization tools such as electron microscopy or scanning probe techniques. In this work, a fully non-invasive, polarization-resolved Raman spectroscopy approach is introduced for determining twist angles in artificially stacked BSCCO heterostructures. By analyzing twist-dependent anisotropic vibrational Raman modes, particularly utilizing the out-of-plane A 1g vibrational mode of Bi/Sr at ≈116 cm −1 , clear optical fingerprints of the rotational misalignment between cuprate layers are identified. The high-resolution confocal Raman setup, equipped with polarization control and RayShield filtering down to 10 cm −1 , allows for reliable and reproducible measurements without compromising the material's structural integrity.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Light Microsopy Module, International Space Station Premier Automated Microscope

The Light Microscopy Module (LMM) was launched to the International Space Station (ISS) in 2009 and began science operations in 2010. It continues to support Physical and Biological scientific research on ISS. During 2015, if all goes as planned, five experiments will be completed: [1] Advanced Colloids Experiments with a manual sample base -3 (ACE-M-3), [2] the Advanced Colloids Experiment with a Heated Base -1 (ACE-H-1), [3] (ACE-H-2), [4] the Advanced Plant Experiment -03 (APEX-03), and [5] the Microchannel Diffusion Experiment (MDE). Preliminary results, along with an overview of present and future LMM capabilities will be presented; this includes details on the planned data imaging processing and storage system, along with the confocal upgrade to the core microscope. [1] New York University: Paul Chaikin, Andrew Hollingsworth, and Stefano Sacanna, [2] University of Pennsylvania: Arjun Yodh and Matthew Gratale, [3] a consortium of universities from the State of Kentucky working through the Experimental Program to Stimulate Competitive Research (EPSCoR): Stuart Williams, Gerold Willing, Hemali Rathnayake, et al., [4] from the University of Florida and CASIS: Anna-Lisa Paul and Rob Ferl, and [5] from the Methodist Hospital Research Institute from CASIS: Alessandro Grattoni and Giancarlo Canavese.

Microscopy↗