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At least 127 records · Page 7

Directional Solidification of Pure Succinonitrile and a Succinonitrile-Acetone Alloy

An experimental study of the horizontal Bridgman growth of pure succinonitrile (SCN) and of a succinonitrile-1.0 mol% acetone alloy (SCN-1.0 mol.% ACE) has been performed. Experiments involving both a stationary thermal field (no-growth case) and a translating thermal field (growth case) were conducted. Growth rates of 2 and 40 micrometers/sec were investigated. For the pure SCN experiments, the velocity field in the melt was estimated using video images of seed particles in the melt. Observations of the seed particles indicate that a primary longitudinal convective cell is formed. The maximum velocity of two different particles which traveled along similar paths was the same and equal to 1.49 +/- 0.01 mm/s. The general accuracy of velocity measurements is estimated to be +/-0.08 mm/s, though the data shows consistency to within +/- 0.02 mm/s. The shape of the solid/liquid interface was also quantitatively determined. The solid/liquid interface was stable (non-dendritic and non-cellular) but not flat: rather it was significantly distorted by the influence of connection in the melt and, for the growth case, by the moving temperature boundary conditions along the ampoule. It was found that the interface shape and position were highly dependent on the alignment of the ampoule in the apparatus. Consequently, the ampoule was carefully aligned for all experiments. The values for front location agree with those determined in previous experiments. For the alloy experiments, the solid/liquid interface was determined to be unstable at growth rates greater than 2.8 micrometers/sec, but stable for the cases of no-growth and growth at 2 micrometers/sec. When compared to the shape of the pure SCN interface, the alloy interface forms closer to the cold zone, indicating that the melting temperature decreased due to the alloying element. Extensive temperature measurements were performed on the outside of the ampoule containing pure SCN. The resulting thermal profiles are presented in detail in the results section. It is intended that the interface shape, thermal boundary condition, and velocity data presented in this paper be used to test numerical simulations.

Simpson, James E.↗

Pluto’s Sputnik Planitia: Composition of geological units from infrared spectroscopy

We have compared spectroscopic data of Sputnik Planitia on Pluto, as acquired by New Horizons’ Linear Etalon Imaging Spectral Array (LEISA) instrument, to the geomorphology as mapped by White et al. (2017) using visible and panchromatic imaging acquired by the LOng-Range Reconnaissance Imager (LORRI) and the Multi-spectral Visible Imaging Camera (MVIC). We have focused on 13 of the geologic units identified by White et al. (2017), which include the plains and mountain units contained within the Sputnik basin. We divided the map of Sputnik Planitia into 15 provinces, each containing one or more geologic units, and we use LEISA to calculate the average spectra of the units inside the 15 provinces. Hapke-based modeling was then applied to the average spectra of the units to infer their surface composition, and to determine if the composition resulting from the modeling of LEISA spectra reflects the geomorphologic analyses of LORRI data, and if areas classified as being the same geologically, but which are geographically separated, share a similar composition. We investigated the spatial distribution of the most abundant ices on Pluto’s surface - CH4, N2, CO, H2O, and a non-ice component presumed to be a macromolecular carbon-rich material, termed a tholin, that imparts a positive spectral slope in the visible spectral region and a negative spectral slope longward of ~1.1 μm. Because the exact nature of the non-ice component is still debated and because the negative spectral slope of the available tholins in the near infrared does not perfectly match the Pluto data, for spectral modeling purposes we reference it generically as the negative spectral slope endmember (NSS endmember). We created maps of variations in the integrated band depth (from LEISA data) and areal mass fraction (from the modeling) of the components. The analysis of correlations between the occurrences of the endmembers in the geologic units led to the observation of an anomalous suppression of the strong CH4 absorption bands in units with compositions that are dominated by H2O ice and the NSS endmember. Exploring the mutual variation of the CH4 and N2 integrated band depths with the abundance of crystalline H2O and NSS endmember revealed that the NSS endmember is primarily responsible for the suppression of CH4 absorptions in mountainous units located along the western edge of Sputnik Planitia. Our spectroscopic analyses have provided additional insight into the geological processes that have shaped Sputnik Planitia. A general increase in volatile abundance from the north to the south of Sputnik Planitia is observed. Such an increase first observed and interpreted by Protopapa et al., 2017 and later confirmed by climate modeling (Bertrand et al., 2018) is expressed geomorphologically in the form of preferential deposition of N2 ice in the upland and mountainous regions bordering the plains of southern Sputnik Planitia. Relatively high amounts of pure CH4 are seen at the southern Tenzing Montes, which are a natural site for CH4 deposition owing to their great elevation and the lower insolation they are presently receiving. The NSS endmember correlates the existence of tholins within certain units, mostly those coating the low-latitude mountain ranges that are co-latitudinal with the tholin-covered Cthulhu Macula. The spectral analysis has also revealed compositional differences between the handful of occurrences of northern non-cellular plains and the surrounding cellular plains, all of which are located within the portion of Sputnik Planitia that is presently experiencing net sublimation of volatiles, and which do not therefore exhibit a surface layer of bright, freshly-deposited N2 ice. The compositional differences between the cellular and non-cellular plains here hint at the effectiveness of convection in entraining and trapping tholins within the body of the cellular plains, while preventing the spread of such tholins to abutting non-cellular plains.

Pluto's Sputnik Planitia↗

The cytoskeleton and gravitropism in higher plants

The cellular and molecular mechanisms underlying the gravitropic response of plants have continued to elude plant biologists despite more than a century of research. Lately there has been increased attention on the role of the cytoskeleton in plant gravitropism, but several controversies and major gaps in our understanding of cytoskeletal involvement in gravitropism remain. A major question in the study of plant gravitropism is how the cytoskeleton mediates early sensing and signal transduction events in plants. Much has been made of the actin cytoskeleton as the cellular structure that sedimenting amyloplasts impinge upon to trigger the downstream signaling events leading to the bending response. There is also strong molecular and biochemical evidence that the transport of auxin, an important player in gravitropism, is regulated by actin. Organizational changes in microtubules during the growth response phase of gravitropism have also been well documented, but the significance of such reorientations in controlling differential cellular growth is unclear. Studies employing pharmacological approaches to dissect cytoskeletal involvement in gravitropism have led to conflicting results and therefore need to be interpreted with caution. Despite the current controversies, the revolutionary advances in molecular, biochemical, and cell biological techniques have opened up several possibilities for further research into this difficult area. The myriad proteins associated with the plant cytoskeleton that are being rapidly characterized provide a rich assortment of candidate regulators that could be targets of the gravity signal transduction chain. Cytoskeletal and ion imaging in real time combined with mutant analysis promises to provide a fresh start into this controversial area of research.

Review↗

Spatial Pattern of Cell Damage in Tissue from Heavy Ions

A new Monte Carlo algorithm was developed that can model passage of heavy ions in a tissue, and their action on the cellular matrix for 2- or 3-dimensional cases. The build-up of secondaries such as projectile fragments, target fragments, other light fragments, and delta-rays was simulated. Cells were modeled as a cell culture monolayer in one example, where the data were taken directly from microscopy (2-d cell matrix). A simple model of tissue was given as abstract spheres with close approximation to real cell geometries (3-d cell matrix), as well as a realistic model of tissue was proposed based on microscopy images. Image segmentation was used to identify cells in an irradiated cell culture monolayer, or slices of tissue. The cells were then inserted into the model box pixel by pixel. In the case of cell monolayers (2-d), the image size may exceed the modeled box size. Such image was is moved with respect to the box in order to sample as many cells as possible. In the case of the simple tissue (3-d), the tissue box is modeled with periodic boundary conditions, which extrapolate the technique to macroscopic volumes of tissue. For real tissue, specific spatial patterns for cell apoptosis and necrosis are expected. The cell patterns were modeled based on action cross sections for apoptosis and necrosis estimated based on BNL data, and other experimental data.

Ponomarev, Artem L.↗

X-ray fluorescence mapping of brain tissue reveals the profound extent of trace element dysregulation in stroke pathophysiology

Abstract The brain is a privileged organ with regard to its trace element composition and maintains a robust barrier system to sequester this specialized environment from the rest of the body and the vascular system. Stroke is caused by loss of adequate blood flow to a region of the brain. Without adequate blood flow ischaemic changes begin almost immediately, triggering an ischaemic cascade, characterized by ion dysregulation, loss of function, oxidative damage, cellular degradation, and breakdown of the barrier that helps maintain this environment. Ion dysregulation is a hallmark of stroke pathophysiology and we observe that most elements in the brain are dysregulated after stroke. X-ray fluorescence-based detection of physiological changes in the neurometallome after stroke reveals profound ion dysregulation within the lesion and surrounding tissue. Not only are most elements significantly dysregulated after stroke, but the level of dysregulation cannot be predicted from a cell-level description of dysregulation. X-ray fluorescence imaging reveals that the stroke lesion retains <25% of essential K+ after stroke, but this element is not concomitantly elevated elsewhere in the organ. Moreover, elements like Na+, Ca2+, and Cl− are vastly elevated above levels available in normal brain tissue (>400%, >200%, and >150%, respectively). We hypothesize that weakening of the blood–brain barrier after stroke allows elements to freely diffuse down their concentration gradient so that the stroke lesion is in equilibrium with blood (and the compartments containing brain interstitial fluid and cerebrospinal fluid). The change observed for the neurometallome likely has consequences for the potential to rescue infarcted tissue, but also presents specific targets for treatment.

Biochemistry & Molecular Biology↗

Unraveling the Dynamics of Nucleosome Arrays

The organization of genomic DNA into chromatin is a fundamental determinant of genome stability, regulation, and cellular function. Nucleosomes, the basic repeating units of chromatin, assemble into higher-order structures whose organization and heterogeneity remain difficult to characterize using conventional ensemble-averaged techniques. A key need in the field is the development of experimental approaches capable of directly visualizing nucleosome assemblies and their structural variability at the single-molecule level. This LDRD Lab-Wide project focused on establishing and evaluating atomic force microscopy (AFM)–based approaches for the characterization of nucleosome assemblies. The work emphasized experimental workflows for preparing, imaging, and assessing multi-nucleosome systems, rather than isolated single nucleosomes. Through method development and exploratory measurements, the project demonstrated the feasibility of applying scanning probe microscopy to investigate chromatin-relevant assemblies and provided preliminary insight into the strengths and limitations of this approach for future quantitative studies. Results and lessons learned from this effort were disseminated to the broader scientific community through multiple national conference presentations, helping to position LLNL for continued work in chromatin and genome organization research.

59 BASIC BIOLOGICAL SCIENCES↗

Impact of Simulated Galactic Cosmic Radiation and Hindlimb Unloading on the Mouse Adrenal Gland Morphology and Histology

Stress related impact of Galactic Cosmic Radiation (GCR) combined with microgravity in space are concerns for astronauts going beyond the Van Allen belts on their way to the Moon and Mars. The adrenal gland is the central endocrine tissue involved in stress management, adrenal hormones regulate the immune system, cellular and tissue metabolism, and glucose and sodium balances. Adult (6 m old) female (n=18) and male (n=18) C57BL/6J mice were divided evenly into 6 treatment groups (control, hindlimb unloading (HU), irradiated (IR), HU+IR, group house control, and group house IR). Adrenals were dissected and fixed for 24 h in 4% paraformaldehyde, followed by transfer to 70% EtOH, and shipment to KUMC. Adrenals were then embedded in paraffin and serial sectioned and 4 central sections from each were stained with H&E, prior to imaging on a Nikon 80i microscope. Whole adrenal cross section area, medullary area, and cortex area were determined. General adrenal gland morphology and histology did not appear to be affected by hindlimb unloading or GCR exposure. Total cross-sectional area, medullary region, and cortex of the adrenal were not different across treatment groups. Male adrenal cross-sectional areas are half the size of the female adrenals, due to a difference in cortex area. In conclusion, simulated microgravity and GCR did not dramatically affect the adrenal gland histology or size. Support from Accelerate Cancer Education Internship Program (KUCC) and KINBRE Summer Scholars Program (KUMC) and NASA Human Research Program (HRP) Human Factors Behavioral Performance Element Grant 18 18FLAG 2 0028

radiation↗

Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape

Studies were carried out with capillary endothelial cells cultured on fibronectin (FN)-coated dishes in order to analyze the mechanism of cell and nuclear shape control by extracellular matrix (ECM). To examine the role of the cytoskeleton in shape determination independent of changes in transmembrane osmotic pressure, membranes of adherent cells were permeabilized with saponin (25 micrograms/ml) using a buffer that maintains the functional integrity of contractile microfilaments. Real-time videomicroscopic studies revealed that addition of 250 microM ATP resulted in time-dependent retraction and rounding of permeabilized cells and nuclei in a manner similar to that observed in intact living cells following detachment using trypsin-EDTA. Computerized image analysis confirmed that permeabilized cells remained essentially rigid in the absence of ATP and that retraction was stimulated in a dose-dependent manner as the concentration of ATP was raised from 10 to 250 microM. Maximal rounding occurred by 30 min with projected cell and nuclear areas being reduced by 69 and 41%, respectively. ATP-induced rounding was also accompanied by a redistribution of microfilaments resulting in formation of a dense net of F-actin surrounding retracted nuclei. Importantly, ATP-stimulated changes in cell, cytoskeletal, and nuclear form were prevented in permeabilized cells using a synthetic myosin peptide (IRICRKG) that has been previously shown to inhibit actomyosin filament sliding in muscle. In contrast, both the rate and extent of cell and nuclear rounding were increased in permeabilized cells exposed to ATP when the soluble FN peptide, GRGDSP, was used to dislodge immobilized FN from cell surface integrin receptors.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center↗

Heavy ion action on single cells: Cellular inactivation capability of single accelerated heavy ions

Heavy ions (HZE-particles) constitute an important part of radiation in space. Although their number is small the high amount of energy transferred by individual particles may cause severe biological effects. Their investigation requires special techniques which were tested by experiments performed at the UNILAC at the GSI (Darmstadt). Diploid yeast was used which is a suitable eucaryotic test system because of its resistance to extreme conditions like dryness and vacuum. Cells were placed on nuclear track detector foils and exposed to ions of different atomic number and energy. To assess the action of one single ion on an individual cell, track parameters and the respective colony forming abilities (CFA) were determined with the help of computer aided image analysis. There is mounting evidence that not only the amount of energy deposited along the particle path, commonly given by the LET, is of importance but also the spatial problem of energy deposition at a submicroscopical scale. It is virtually impossible to investigate track structure effects in detail with whole cell populations and (globally applied) high particle fluences. It is, therefore, necessary to detect the action of simple ions in individual cells. The results show that the biological action depends on atomic number and specific energy of the impinging ions, which can be compared with model calculations of recent track structure models.

Kost, M.↗

Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions

We have described a powerful tool for the study of mechanical interactions between cells and their physical environment. Although the approach has already been used in a variety of ways to measure traction forces and to characterize active and passive responses of cultured cells to mechanical stimulation, it can be extended easily and combined with other microscopic approaches, including fluorescent analog imaging (Beningo et al., 2001), photobleaching, calcium imaging, micromanipulation, and electrophysiology. This method will be particularly useful for studying the functions of various components at focal adhesions, and the effects of mechanical forces on focal adhesion-mediated signal transduction. In addition, the method can be extended to a 3D setting, e.g., by sandwiching cultured cells between two layers of polyacrylamide to create an environment mimicking that in the tissue of a multicellular organism. Whereas chemical interactions between cells and the environment have been investigated extensively, many important questions remain as to the role of physical forces in cellular functions and the interplay between chemical and physical mechanisms of communication. The present approach, as well as other approaches capable of probing physical interactions, should fill in this important gap in the near future.

NASA Discipline Cell Biology↗

A regional ultrastructural analysis of the cellular and synaptic architecture in the chinchilla cristae ampullares

The chinchilla crista ampullaris was studied in 10 samples, each containing 32 consecutive ultrathin sections of the entire neuroepithelium. Dissector methods were used to estimate the incidence of various synaptic features, and results were confirmed in completely reconstructed hair cells. There are large regional variations in cellular and synaptic architecture. Type I and type II hair cells are shorter, broader, and less densely packed in the central zone than in the intermediate and peripheral zones. Complex calyx endings are most common centrally. On average, there are 15-20 ribbon synapses and 25-30 calyceal invaginations in each type I hair cell. Synapses and invaginations are most numerous centrally. Central type II hair cells receive considerably fewer afferent boutons than do peripheral type II hair cells, but have similar numbers of ribbon synapses. The numbers are similar because central type II hair cells make more synapses with the outer faces of calyx endings and with individual afferent boutons. Most afferent boutons get one ribbon synapse. Boutons without ribbon synapses were only found peripherally, and boutons getting multiple synapses were most frequent centrally. Throughout the neuroepithelium, there is an average of three to four efferent boutons on each type II hair cell and calyx ending. Reciprocal synapses are rare. Most synaptic ribbons in type I hair cells are spherules; those in type II hair cells can be spherical or elongated and are particularly heterogeneous centrally. Consistent with the proposal that the crista is concentrically organized, the intermediate and peripheral zones are each similar in their cellular and synaptic architecture near the base and near the planum. An especially differentiated subzone may exist in the middle of the central zone.

Non-NASA Center↗

Electron microscopy evidence of gadolinium toxicity being mediated through cytoplasmic membrane dysregulation

Past functional toxicogenomic studies have indicated that genes relevant to membrane lipid synthesis are important for tolerance to the lanthanides. Moreover, previously reported imaging of patient's brains following administration of gadolinium-based contrast agents shows gadolinium lining the vessels of the brain. Taken together, these findings suggest the disruption of cytoplasmic membrane integrity as a mechanism by which lanthanides induce cytotoxicity. In the presented work we used scanning transmission electron microscopy and spatially resolved elemental spectroscopy to image the morphology and composition of gadolinium, europium, and samarium precipitates that formed on the outside of yeast cell membranes. In no sample did we find that the lanthanide contaminant had crossed the cell membrane, even in experiments using yeast mutants with disrupted genes for sphingolipid synthesis—the primary lipids found in yeast cytoplasmic membranes. Rather, we have evidence that lanthanides are co-located with phosphorus outside the yeast cells. Finally, these results lead us to hypothesize that the lanthanides scavenge or otherwise form complexes with phosphorus from the sphingophospholipid head groups in the cellular membrane, thereby compromising the structure or function of the membrane, and gaining the ability to disrupt membrane function without entering the cell.

59 BASIC BIOLOGICAL SCIENCES↗

White-light movies of the solar photosphere from the SOUP instrument on Spacelab

Initial results are presented on solar granulation, pores and sunspots from the white-light films obtained by the Solar Optical Universal Polarimeter (SOUP) instrument in Spacelab 2. Several hours of movies were taken at various disk and limb positions in quiet and active regions. The images are diffraction-limited at 0.5 arcsec resolution and are, of course, free of atmospheric seeing and distortion. Properties of the granulation in magnetic and nonmagnetic regions are compared and are found to differ significantly in size, rate of intensity variation, and lifetime. In quiet sun, on the order of fifty-percent of the area has at least one 'exploding granule' occurring in it during a 25-min period. Local correlation tracking has detected several types of transverse flows, including systematic outflow from the penumbral boundary of a spot, motion of penumbral filaments, and cellular flow patterns of supergranular and mesogranular size. Feature tracking has shown that, in the quiet sun, the average granule fragment has a velocity of about one kilometer/second.

Title, A. M.↗

Regulation of Absorption and Emission in a Protein/Fluorophore Complex

Human cellular retinol binding protein II (hCRBPII) was used as a protein engineering platform to rationally regulate absorptive and emissive properties of a covalently bound fluorogenic dye. We demonstrate the binding of a thio-dapoxyl analog via formation of a protonated imine between an active site lysine residue and the chromophore’s aldehyde. Rational manipulation of the electrostatics of the binding pocket results in a 204 nm shift in absorption and a 131 nm shift in emission. The protein is readily expressed in mammalian systems and binds with exogenously delivered fluorophore as demonstrated by live-cell imaging experiments.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The three-dimensional structure of cumulus clouds over the ocean. 1: Structural analysis

Thermal channel (channel 6, 10.4-12.5 micrometers) images of five Landsat thematic mapper cumulus scenes over the ocean are examined. These images are thresholded using the standard International Satellite Cloud Climatology Project (ISCCP) thermal threshold algorithm. The individual clouds in the cloud fields are segmented to obtain their structural statistics which include size distribution, orientation angle, horizontal aspect ratio, and perimeter-to-area (PtA) relationship. The cloud size distributions exhibit a double power law with the smaller clouds having a smaller absolute exponent. The cloud orientation angles, horizontal aspect ratios, and PtA exponents are found in good agreement with earlier studies. A technique also is developed to recognize individual cells within a cloud so that statistics of cloud cellular structure can be obtained. Cell structural statistics are computed for each cloud. Unicellular clouds are generally smaller (less than or equal to 1 km) and have smaller PtA exponents, while multicellular clouds are larger (greater than or equal to 1 km) and have larger PtA exponents. Cell structural statistics are similar to those of the smaller clouds. When each cell is approximated as a quadric surface using a linear least squares fit, most cells have the shape of a hyperboloid of one sheet, but about 15% of the cells are best modeled by a hyperboloid of two sheets. Less than 1% of the clouds are ellipsoidal. The number of cells in a cloud increases slightly faster than linearly with increasing cloud size. The mean nearest neighbor distance between cells in a cloud, however, appears to increase linearly with increasing cloud size and to reach a maximum when the cloud effective diameter is about 10 km; then it decreases with increasing cloud size. Sensitivity studies of threshold and lapse rate show that neither has a significant impact upon the results. A goodness-of-fit ratio is used to provide a quantitative measure of the individual cloud results. Significantly improved results are obtained after applying a smoothing operator, suggesting the eliminating subresolution scale variations with higher spatial resolution may yield even better shape analyses.

Kuo, Kwo-Sen↗

Mapping and Quantification of Vascular Branching in Plants, Animals and Humans by VESGEN Software

Humans face daunting challenges in the successful exploration and colonization of space, including adverse alterations in gravity and radiation. The Earth-determined biology of plants, animals and humans is significantly modified in such extraterrestrial environments. One physiological requirement shared by larger plants and animals with humans is a complex, highly branching vascular system that is dynamically responsive to cellular metabolism, immunological protection and specialized cellular/tissue function. VESsel GENeration (VESGEN) Analysis has been developed as a mature beta version, pre-release research software for mapping and quantification of the fractal-based complexity of vascular branching. Alterations in vascular branching pattern can provide informative read-outs of altered vascular regulation. Originally developed for biomedical applications in angiogenesis, VESGEN 2D has provided novel insights into the cytokine, transgenic and therapeutic regulation of angiogenesis, lymphangiogenesis and other microvascular remodeling phenomena. Vascular trees, networks and tree-network composites are mapped and quantified. Applications include disease progression from clinical ophthalmic images of the human retina; experimental regulation of vascular remodeling in the mouse retina; avian and mouse coronary vasculature, and other experimental models in vivo. We envision that altered branching in the leaves of plants studied on ISS such as Arabidopsis thaliana cans also be analyzed.

Parsons-Wingerter, P. A.↗

Mapping and Quantification of Vascular Branching in Plants, Animals and Humans by VESGEN Software

Humans face daunting challenges in the successful exploration and colonization of space, including adverse alterations in gravity and radiation. The Earth-determined biology of humans, animals and plants is significantly modified in such extraterrestrial environments. One physiological requirement shared by humans with larger plants and animals is a complex, highly branching vascular system that is dynamically responsive to cellular metabolism, immunological protection and specialized cellular/tissue function. The VESsel GENeration (VESGEN) Analysis has been developed as a mature beta version, pre-release research software for mapping and quantification of the fractal-based complexity of vascular branching. Alterations in vascular branching pattern can provide informative read-outs of altered vascular regulation. Originally developed for biomedical applications in angiogenesis, VESGEN 2D has provided novel insights into the cytokine, transgenic and therapeutic regulation of angiogenesis, lymphangiogenesis and other microvascular remodeling phenomena. Vascular trees, networks and tree-network composites are mapped and quantified. Applications include disease progression from clinical ophthalmic images of the human retina; experimental regulation of vascular remodeling in the mouse retina; avian and mouse coronary vasculature, and other experimental models in vivo. We envision that altered branching in the leaves of plants studied on ISS such as Arabidopsis thaliana cans also be analyzed.

Parsons-Wingerter, Patricia A.↗

Spatial Distribution and Clustering of Glycosaminoglycans in Electrospun Gelatin-Based Scaffolds

The extracellular matrix (ECM) is comprised of components like collagen, elastin, and glycosaminoglycans (GAGs). Electrospun fibrous scaffolds are designed to replicate the form and composition of the native ECM, often requiring blending of various ECM component mimics to enhance cellular responses. However, the spatial distribution of blended components within these fibers remains unclear. This study investigates the spatial distribution of chondroitin sulfate-C (CSC) in electrospun gelatin-based scaffolds. scanning electron microscopy (SEM), attenuated reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and Time-of-flight Secondary Ion Mass Spectrometry (ToF-SIMS) were applied for surface and subsurface chemical characterization of the fibrous scaffolds. SEM confirmed a fibrous morphology, while ATR-FTIR and XPS analyses indicated the presence of CSC through the identification of sulfate groups. ToF-SIMS imaging, alongside K-means clustering and Ripley’s K function, revealed a nonuniform CSC distribution with higher concentrations at the top layer of the scaffold. This study demonstrates that CSC presentation at the fiber surface varies with depth and differs from bulk incorporation while reveals nanoscale clustering and spatial heterogeneity at both the surface and subsurface of electrospun gelatin fibers. These findings define an underexplored design consideration with potential to influence cell–scaffold interactions.

Animal derived food↗