Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “FLUORESCEIN”

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.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 91 records · Page 5
All sourcesNASA NTRSDOE OSTI
DOE OSTI2017

Synthesis of two fluorescent GTP$\gamma$S molecules and their biological relevance

Fluorescent GTP analogues are utilized for an assortment of nucleic acid and protein characterization studies. Non-hydrolysable analogues such as GTPγS offer the advantage of keeping proteins in a GTP-bound conformation due to their resistance to hydrolysis into GDP. Two novel fluorescent GTPγS molecules were developed by linking fluorescein and tetramethylrhodamine to the γ-thiophosphate of GTPγS. Chemical and biological analysis of these two compounds revealed their successful synthesis and ability to bind to the nucleotide-binding site of tubulin. These two new fluorescent non-hydrolysable nucleotides offer new possibilities for biophysical and biochemical characterization of GTP-binding proteins.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2018

Aging Exacerbates Neuroinflammatory Outcomes Induced by Acute Ozone Exposure

The role of environmental stressors, particularly exposure to air pollution, in the development of neurodegenerative disease remains underappreciated. We examined the neurological effects of acute ozone (O 3 ) exposure in aged mice, where increased blood brain barrier (BBB) permeability may confer vulnerability to neuroinflammatory outcomes. C 57 BL/6 male mice, aged 8-10 weeks or 12–18 months were exposed to either filtered air (FA) or 1.0 ppm O 3 for 4 hours; animals received a single IP injection of sodium fluorescein (FSCN) 20 hours post-exposure. One-hour post-FSCN injection, animals were transcardially perfused for immunohistochemical analysis of BBB permeability. β-amyloid protein expression was assessed via ELISA. Flow cytometric characterization of infiltrating immune cells, including neutrophils, macrophages, and microglia populations was performed 20 hours post-O 3 exposure. Flow cytometry analysis of brains revealed increased microglia “activation” and presentation of CD11b, F4/80 and MHCII in aged animals relative to younger ones; these age-induced differences were potentiated by acute O 3 exposure. Cortical and limbic regions in aged brains had increased reactive microgliosis and β-amyloid protein expression after O 3 insult. The aged cerebellum was particularly vulnerable to acute O 3 exposure with increased populations of infiltrating neutrophils, peripheral macrophages/monocytes, and Ly6C + inflammatory monocytes after insult, which were not significantly increased in the young cerebellum. O 3 exposure increased the penetration of FSCN beyond the BBB, the infiltration of peripheral immune cells, and reactive gliosis of microglia. Furthermore, the aged BBB is vulnerable to insult and becomes highly penetrable in response to O 3 exposure, leading to greater neuroinflammatory outcomes.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2017

Energy transfer networks: Quasicontinuum photoluminescence linked to high densities of defects

In a series of studies related to laser-induced damage of optical materials and deposition of plastics, we discovered a broadly emitting photoluminescence with fast lifetimes that we termed quasicontinuum photoluminescence (QC-PL). Here in this paper, we suggest that a high density of optically active defects leads to QC-PL, where interactions between defects affect the temporal and spectral characteristics of both excitation and emission. We develop a model that predicts the temporal characteristics of QC-PL, based on energy transfer interactions between high densities of defects. Our model does not explain all spectral broadening and redshifts found in QC-PL, since we do not model spectral changes in defects due to proximity to other defects. However, we do provide an example of a well-defined system that exhibits the QC-PL characteristics of a distribution in shortened lifetimes and broadened, redshifted energy levels: an organic chromophore (fluorescein) that has been dried rapidly on a fused silica surface. Recently, we showed that regions of fused silica exposed to up to 1 billion high-fluence laser shots at 351 rm nm at subdamage fluences exhibit significant transmission losses at the surface. Here, we find that these laser-exposed regions also exhibit QC-PL. Increases in the density of induced defects on these laser-exposed surfaces, as measured by the local transmission loss, lead to decreases in the observed lifetime and redshifts in the spectrum of the QC-PL, consistent with our explanation for QC-PL. In conclusion, we have found QC-PL in an increasing variety of situations and materials, and we believe it is a phenomenon commonly found on surfaces and nanostructured materials.

36 MATERIALS SCIENCE↗
DOE OSTI2016

TU-H-CAMPUS-TeP2-04: Measurement of Stereotactic Output Factors with DNA Double-Strand Breaks

Purpose: Radiotherapy treatment is specified by radiation dose prescriptions, but biological DNA damage actually controls treatment effectiveness. It is impractical to directly measure dose in the clinic, so we measure quantities, such as collected charge, and calculate the relationship to dose. At small fields, such as those in stereotactic radiosurgery (SRS), charged-particle equilibrium (CPE) breaks down and the accuracy of the measurement for delivered dose decreases. By measuring DNA double-strand breaks (DSB) directly, we believe treatment accuracy could improve by providing a more meaningful measurement. Methods: A DNA dosimeter, consisting of magnetic streptavidin beads attached to 4 kilobase pair DNA strands labeled with biotin and fluorescein amidite (FAM) on opposing ends, was suspended in phosphate-buffered saline (PBS). Twenty µL samples were placed in plastic micro-capillary tubes inside a water tank setup and irradiated with 10 cm, 3 cm, 1.25 cm, 0.75 cm, and 0.5 cm radiation field sizes, where the three smallest sizes were cones. After irradiation, the dosimeters were mechanically separated into beads (intact DNA) and supernatant (broken DNA/FAM) using a magnet. The fluorescence was read and the probability of DSB was calculated. This was used to calculate the output factor for an SRS beam and compared to that measured using a diode detector. Results: The output factors relative to a 10 cm field were 0.89±0.07, 0.76±0.08, 0.59±0.04, and 0.78±0.12 for the field sizes of 3 cm, 1.25 cm, 0.75 cm, and 0.5 cm, respectively. Some of the diode measurements do not fall within these uncertainties. Conclusion: This was the first attempt to measure output factors in a water tank with the DNA dosimeter. Although differences compared to the diode were observed, the uncertainty analysis ignored systematic errors. For future work, we will repeat this experiment to quantify and correct systematic errors, such as those caused by positional alignment and sample contamination. This work was funded in part by CPRIT (RP140105).

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

TH-CD-201-11: Optimizing the Response and Cost of a DNA Double-Strand Break Dosimeter

Purpose: A DNA double-strand break (DSB) dosimeter was developed to measure the biological effect of radiation. The goal here is to refine the fabrication method of this dosimeter to reproducibly create a low coefficient of variation (CoV) and reduce the cost for the dosimeter. Methods: Our dosimeter consists of 4 kilo-base pair DNA strands (labeled on one end with biotin and on the other with fluorescein) attached to streptavidin magnetic beads. The final step of the DNA dosimeter fabrication is to suspend these attached beads in phosphate-buffered saline (PBS). The amount of PBS used to suspend the attached beads and the relative volume of the DNA strands to the beads both affect the CoV and dosimeter cost. We diluted the beads attached with DNA in different volumes of PBS (100, 200, and 400 µL) to create different concentrations of the DNA dosimeter. Then we irradiated these dosimeters (50 µL samples) in a water-equivalent plastic phantom at 25 and 50 Gy (three samples per dose) and calculated the CoV for each dosimeter concentration. Also, we used different masses of DNA strands (1, 2, 8, 16, 24, and 32 µg) to attach to the same volume of magnetic beads (100 µL) to explore how this affects the cost of the dosimeter. Results: The lowest CoV was produced for the highest concentration of dosimeter (100 µL of PBS), which created CoV of 2.0 and 1.0% for 25 and 50 Gy, respectively. We found that the lowest production cost for the dosimeter occurs by attaching 16 µg of DNA strands with 100 µL of beads. Conclusion: : We optimized the fabrication of the DNA dosimeter to produce low CoV and cost, but we still need to explore ways to further improve the dosimeter for use at lower doses. This work was supported in part by Yarmouk University (Irbid, Jordan) and CPRIT (RP140105)

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Соllective Fluorescence of Composite Nanoparticles

Fluorescence of a suspension of spherical nanoparticles consisting of a gold core surrounded by silicon dioxide doped with fluorescein molecules is experimentally studied. The model of a composite nanoparticle is investigated theoretically and experimentally, taking into account polarization fluctuations. It is shown that a local nonlinear feedback in the system leads to characteristic temperature dependences of the fluorescence linewidth and intensity. As the medium was cooled from room temperature to liquid nitrogen temperature, the fluorescence spectrum narrowed and its intensity strongly increased. A comparison of experimental data with numerical calculations showed that the changes observed in experiments are not explained by the temperature dependence of the parameters of elements of a nanoparticle. The analysis of the dynamics of polarization phases of dye molecules showed that the synergetic effect should be taken into account, which forms the basis of plasmon–polariton superradiance.

36 MATERIALS SCIENCE↗
DOE OSTI2018

Micropuncture of Bowman's Space in Mice Facilitated by 2 Photon Microscopy

Renal micropuncture and renal 2-photon imaging are seminal techniques in renal physiology. However, micropuncture is limited by dependence on conventional microscopy to surface nephron features, and 2-photon studies are limited in that interventions can only be assessed at the organ, rather than the nephron level. In particular, micropuncture studies of the glomeruli of mice have been challenged by the paucity of surface glomeruli in mice. To address this limitation in order to pursue studies of aspirate from Bowman's space in mouse physiologic models, we developed 2-photon glomerular micropuncture. We present a novel surgical preparation that allows lateral access to the kidney while preserving the required vertical imaging column for 2-photon microscopy. Administration of high molecular weight fluorescein isothiocyanate (FITC)- dextran is used to render the renal vasculature and therefore glomeruli visible for 2-photon imaging. A quantum dot-coated pipette is then introduced under stereotactic guidance to a glomerulus selected from the several to many which may be visualized within the imaging window. In this protocol, we provide details of the preparation, materials, and methods necessary to carry out the procedure. This technique facilitates previously-impossible physiologic study of the kidney, including recovery of filtrate from Bowman's space and all segments of the nephron within the imaging depth limit, about 100 µm below the renal capsule. Pressure, charge and flow may all be measured using the introduced pipette. Here, we provide representative data from liquid chromatography/mass spectrometry performed on aspirate from Bowman's space. We expect this technique to have wide applicability in renal physiologic investigation.

Matsushita, Katsuyuki↗
DOE OSTI2023

Using Vertically Aligned Carbon Nanofiber Arrays on Rigid or Flexible Substrates for Delivery of Biomolecules and Dyes to Plants

The delivery of biomolecules and impermeable dyes to intact plants is a major challenge. Nanomaterials are up-and-coming tools for the delivery of DNA to plants. As exciting as these new tools are, they have yet to be widely applied. Nanomaterials fabricated on rigid substrate (backing) are particularly difficult to successfully apply to curved plant structures. This study describes the process for microfabricating vertically aligned carbon nanofiber arrays and transferring them from a rigid to a flexible substrate. We detail and demonstrate how these fibers (on either rigid or flexible substrates) can be used for transient transformation or dye (e.g., fluorescein) delivery to plants. We show how VACNFs can be transferred from rigid silicon substrate to a flexible SU-8 epoxy substrate to form flexible VACNF arrays. Further, to overcome the hydrophobic nature of SU-8, fibers in the flexible film were coated with a thin silicon oxide layer (2-3 nm). To use these fibers for delivery to curved plant organs, we deposit a 1 µL droplet of dye or DNA solution on the fiber side of VACNF films, wait 10 min, place the films on the plant organ and employ a swab with a rolling motion to drive fibers into plant cells. With this method, we have achieved dye and DNA delivery in plant organs with curved surfaces.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1972

Cine recording ophthalmoscope

Camera system provides accurate photographic recording during acceleration of centrifuge and permits immediate observation of dynamic changes in retinal circulation by a closed-circuit television loop. System consists of main camera, remote control unit, and strobe power supply unit, and is used for fluorescein studies and dynamometry sequences.

Fitzgerald, J. W.↗
NASA NTRS1972

Genetic and metabolic variability in autotrophic and heterotrophic bacteria

The studies to evaluate an organism's ability to maintain normal physiological activities over a long period of time in a bioregenerative system are presented. Studies reviewed include: heat tolerant mutants of Pseudomonas fluoresceins, virulence factors of the Staphylococci, and the effect of mutations on the virulence for man in common and ubiquitous microorganisms.

Decicco, B. T.↗
NASA NTRS1974

Tracking air-dropped drogues and dyes from aircraft in support of ERTS-1 circulation studies

The author has identified the following significant results. For two years ERTS-1 has been employed to investigate current circulation patterns in Delaware Bay under different tidal, flow, and wind conditions. Since sufficient numbers of current meters and boats are not available, air-droppable drogues and dye packs have been developed and tested. The drogues consist of a styrofoam float and a line to which is attached a stainless steel biplane. The length of the line determines at what depth currents will be monitored. The floats are color coded to distinguish their movement and mark the depth of the biplanes. Simultaneously floating and anchored dye packs of fluorescein dye have been deployed from aircraft. The movement of the dye and drogues is tracked by sequential aerial photography, using fixed markers on shore or on buoys as reference points to calibrate the scale and direction of drogue movement. The current data obtained by this technique is then used to annotate current circulation maps derived from ERTS-1 imagery.

Klemas, V.↗
NASA NTRS1976

Flight 1 technical report for experiment 74-37 contained polycrystalline solidification in low-G

A .005 M solution of fluorescein in cyclohexanol was directionally solidified in a standard 10 x 10 x 45mm UV silica cuvette, using a bottom thermoelectric chilling device. Progress of the experiment was monitored by time lapse photography. During flight (SPAR I) the camera malfunctioned and only one quarter of the expected data were collected. Comparison of flight and ground specimens indicated that: (1) The dark green layer observed ahead of the solid-liquid interface which is most likely the solute-enriched zone, appears to be wider in the flight specimen; (2) Parasitic nucleation ahead of the solid-liquid interface in the flight sample led to an irregularly shaped interface, smaller grain size, equiaxed grain morphology and a larger average macroscopic growth rate; (3) The formation of equiaxed grains ahead of the solid-liquid interface in the flight specimen may be attributed to ordered islands within the liquid, which survived remelting because of the low degree of superheating (approximately equal to 1.5 C), did not settle because of reduced gravity and acted as nuclei during cooling.

Papaziak, J. M.↗
NASA NTRS1978

Outfall siting with dye-buoy remote sensing of coastal circulation

A dye-buoy remote sensing technique has been applied to estuarine siting problems that involve fine-scale circulation. Small hard cakes of sodium fluorescein and polyvinyl alcohol, in anchored buoys and low-windage current followers, dissolve to produce dye marks resolvable in 1:60,000 scale color and color infrared imagery. Lagrangian current vectors are determined from sequential photo coverage. Careful buoy placement reveals surface currents and submergence near fronts and convergence zones. The technique has been used in siting two sewage outfalls in Hampton Roads, Virginia: In case one, the outfall region during flood tide gathered floating materials in a convergence zone, which then acted as a secondary source during ebb; for better dispersion during ebb, the proposed outfall site was moved further offshore. In case two, flow during late flood was found to divide, with one half passing over shellfish beds; the proposed outfall site was consequently moved to keep effluent in the other half.

Munday, J. C., Jr.↗
NASA NTRS1983

Flow visualization studies of bodies with square cross sections

A water-tunnel study was conducted of four bodies. A solution of sodium fluorescein coating the body provided visualization of vortices and feeding sheets and isolated dots of methyl blue dye provided visualization of stream lines. These data, along with published oil-flow photos, were analyzed to develop the topological representation of the flows in cross-flow planes. Presented are the development of the flow along the body at fixed angles of attack and at a fixed body station with changes in angle of attack. Effects of roll angle, body corner radius, and nose bluntness are illustrated.

Chapman, G. T.↗
NASA NTRS1982

Characteristics of statoliths from rootcaps and coleoptiles

Amyloplasts with intact envelopes were isolated from corn coleoptiles, incubated with either cationized ferritin (CF) or fluorescein isothiocyanate (FITC)-CF, and examined with epifluorescence optics. Before incubation, the mean zeta potential was found to be -19.4 mv. The net negative surface charge was confirmed ultrastructurally by the binding of CF to both amyloplasts and starch grains. Binding to the latter was reduced compared to the former. FITC-CF showed binding to some but not all amyloplasts, and was absent from individual starch granules. Membrane-bound calcium, indicated by chlorotetracycline, fluorescence, was detected throughout the cytoplasm of root cap cells. These results may reflect an affinity of CF for membrane-based charge, and suggest that a calcium shell exists on the amyloplast envelope.

Sack, F. D.↗
NASA NTRS1985

Evidence that Resorption of Bone by Rat Peritoneal Macrophages Occurs in an Acidic Environment

Skeletal loss in space, like any form of osteoporosis, reflects a relative imbalance of the activities of cells resorbing (degrading) or forming bone. Consequently, prevention of weightlessness induced bone loss may theoretically be accomplished by (1) stimulating bone formation or (2) inhibiting bone resorption. This approach, however, requires fundamental understanding of the mechanisms by which cells form or degrade bone, information not yet at hand. An issue central to bone resorption is the pH at which resorption takes place. The pH dependent spectral shift of a fluorescent dye (fluorescein isothiocyanate) conjugated to bone matrix was used to determine the pH at the resorptive cell bone matrix interface. Devitalized rat bone was used as the substrate, and rat peritoneal macrophages were used as the bone resorbing cells. The results suggest that bone resorption is the result of generation of an acidic microenvironment at the cell matrix junction.

Blair, H. C.↗
NASA NTRS1985

Characterization of flow in a scroll duct

A quantitative, flow visualization study was made of a partially elliptic cross section, inward curving duct (scroll duct), with an axial outflow through a vaneless annular cutlet. The working fluid was water, with a Re(d) of 40,000 at the inlet to the scroll duct, this Reynolds number being representative of the conditions in an actual gas turbine scroll. Both still and high speed moving pictures of fluorescein dye injected into the flow and illuminated by an argon ion laser were used to document the flow. Strong secondary flow, similar to the secondary flow in a pipe bend, was found in the bottom half of the scroll within the first 180 degs of turning. The pressure field set up by the turning duct was strong enough to affect the inlet flow condition. At 90 degs downstream, the large scale secondary flow was found to be oscillatory in nature. The exit flow was nonuniform in the annular exit. By 270 degs downstream, the flow appeared unorganized with no distinctive secondary flow pattern. Large scale structures from the upstream core region appeared by 90 degs and continued through the duct to reenter at the inlet section.

Begg, E. K.↗