Microplate-compatible total internal reflection fluorescence microscopy for receptor pharmacology
NASA Astrophysics Data System (ADS)
Chen, Minghan; Zaytseva, Natalya V.; Wu, Qi; Li, Min; Fang, Ye
2013-05-01
We report the use of total internal reflection fluorescence (TIRF) microscopy for analyzing receptor pharmacology and the development of a microplate-compatible TIRF imaging system. Using stably expressed green fluorescence protein tagged β2-adrenergic receptor as the reporter, we found that the activation of different receptors results in distinct kinetic signatures of the TIRF intensity of cells. These TIRF signatures closely resemble the characteristics of their respective label-free dynamic mass redistribution signals in the same cells. This suggests that TIRF in microplate can be used for profiling and screening drugs.
Chung, Euiheon; Kim, Daekeun; Cui, Yan; Kim, Yang-Hyo; So, Peter T. C.
2007-01-01
The development of high resolution, high speed imaging techniques allows the study of dynamical processes in biological systems. Lateral resolution improvement of up to a factor of 2 has been achieved using structured illumination. In a total internal reflection fluorescence microscope, an evanescence excitation field is formed as light is total internally reflected at an interface between a high and a low index medium. The <100 nm penetration depth of evanescence field ensures a thin excitation region resulting in low background fluorescence. We present even higher resolution wide-field biological imaging by use of standing wave total internal reflection fluorescence (SW-TIRF). Evanescent standing wave (SW) illumination is used to generate a sinusoidal high spatial frequency fringe pattern on specimen for lateral resolution enhancement. To prevent thermal drift of the SW, novel detection and estimation of the SW phase with real-time feedback control is devised for the stabilization and control of the fringe phase. SW-TIRF is a wide-field superresolution technique with resolution better than a fifth of emission wavelength or ∼100 nm lateral resolution. We demonstrate the performance of the SW-TIRF microscopy using one- and two-directional SW illumination with a biological sample of cellular actin cytoskeleton of mouse fibroblast cells as well as single semiconductor nanocrystal molecules. The results confirm the superior resolution of SW-TIRF in addition to the merit of a high signal/background ratio from TIRF microscopy. PMID:17483188
A Platform for Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence
Asanov, Alexander; Zepeda, Angélica; Vaca, Luis
2012-01-01
We have developed a novel microarray technology based on total internal reflection fluorescence (TIRF) in combination with DNA and protein bioassays immobilized at the TIRF surface. Unlike conventional microarrays that exhibit reduced signal-to-background ratio, require several stages of incubation, rinsing and stringency control, and measure only end-point results, our TIRF microarray technology provides several orders of magnitude better signal-to-background ratio, performs analysis rapidly in one step, and measures the entire course of association and dissociation kinetics between target DNA and protein molecules and the bioassays. In many practical cases detection of only DNA or protein markers alone does not provide the necessary accuracy for diagnosing a disease or detecting a pathogen. Here we describe TIRF microarrays that detect DNA and protein markers simultaneously, which reduces the probabilities of false responses. Supersensitive and multiplexed TIRF DNA and protein microarray technology may provide a platform for accurate diagnosis or enhanced research studies. Our TIRF microarray system can be mounted on upright or inverted microscopes or interfaced directly with CCD cameras equipped with a single objective, facilitating the development of portable devices. As proof-of-concept we applied TIRF microarrays for detecting molecular markers from Bacillus anthracis, the pathogen responsible for anthrax. PMID:22438738
New light on ion channel imaging by total internal reflection fluorescence (TIRF) microscopy.
Yamamura, Hisao; Suzuki, Yoshiaki; Imaizumi, Yuji
2015-05-01
Ion channels play pivotal roles in a wide variety of cellular functions; therefore, their physiological characteristics, pharmacological responses, and molecular structures have been extensively investigated. However, the mobility of an ion channel itself in the cell membrane has not been examined in as much detail. A total internal reflection fluorescence (TIRF) microscope allows fluorophores to be imaged in a restricted region within an evanescent field of less than 200 nm from the interface of the coverslip and plasma membrane in living cells. Thus the TIRF microscope is useful for selectively visualizing the plasmalemmal surface and subplasmalemmal zone. In this review, we focused on a single-molecule analysis of the dynamic movement of ion channels in the plasma membrane using TIRF microscopy. We also described two single-molecule imaging techniques under TIRF microscopy: fluorescence resonance energy transfer (FRET) for the identification of molecules that interact with ion channels, and subunit counting for the determination of subunit stoichiometry in a functional channel. TIRF imaging can also be used to analyze spatiotemporal Ca(2+) events in the subplasmalemma. Single-molecule analyses of ion channels and localized Ca(2+) signals based on TIRF imaging provide beneficial pharmacological and physiological information concerning the functions of ion channels. Copyright © 2015 The Authors. Production and hosting by Elsevier B.V. All rights reserved.
Axial superresolution via multiangle TIRF microscopy with sequential imaging and photobleaching
Fu, Yan; Winter, Peter W.; Rojas, Raul; Wang, Victor; McAuliffe, Matthew; Patterson, George H.
2016-01-01
We report superresolution optical sectioning using a multiangle total internal reflection fluorescence (TIRF) microscope. TIRF images were constructed from several layers within a normal TIRF excitation zone by sequentially imaging and photobleaching the fluorescent molecules. The depth of the evanescent wave at different layers was altered by tuning the excitation light incident angle. The angle was tuned from the highest (the smallest TIRF depth) toward the critical angle (the largest TIRF depth) to preferentially photobleach fluorescence from the lower layers and allow straightforward observation of deeper structures without masking by the brighter signals closer to the coverglass. Reconstruction of the TIRF images enabled 3D imaging of biological samples with 20-nm axial resolution. Two-color imaging of epidermal growth factor (EGF) ligand and clathrin revealed the dynamics of EGF-activated clathrin-mediated endocytosis during internalization. Furthermore, Bayesian analysis of images collected during the photobleaching step of each plane enabled lateral superresolution (<100 nm) within each of the sections. PMID:27044072
Boehm, Elizabeth M.; Subramanyam, Shyamal; Ghoneim, Mohamed; Washington, M. Todd; Spies, Maria
2016-01-01
Large, dynamic macromolecular complexes play essential roles in many cellular processes. Knowing how the components of these complexes associate with one another and undergo structural rearrangements is critical to understanding how they function. Single-molecule total internal reflection fluorescence (TIRF) microscopy is a powerful approach for addressing these fundamental issues. In this article, we first discuss single-molecule TIRF microscopes and strategies to immobilize and fluorescently label macromolecules. We then review the use of single-molecule TIRF microscopy to study the formation of binary macromolecular complexes using one-color imaging and inhibitors. We conclude with a discussion of the use of TIRF microscopy to examine the formation of higher-order (i.e., ternary, quaternary, etc.) complexes using multi-color setups. The focus throughout this article is on experimental design, controls, data acquisition, and data analysis. We hope that single-molecule TIRF microscopy, which has largely been the province of specialists, will soon become as common in the tool box of biophysicists and biochemists as structural approaches has become today. PMID:27793278
Byrne, Gerard D; Vllasaliu, Driton; Falcone, Franco H; Somekh, Michael G; Stolnik, Snjezana
2015-11-02
In this work we utilize the combination of label-free total internal reflection microscopy and total internal reflectance fluorescence (TIRM/TIRF) microscopy to achieve a simultaneous, live imaging of single, label-free colloidal particle endocytosis by individual cells. The TIRM arm of the microscope enables label free imaging of the colloid and cell membrane features, while the TIRF arm images the dynamics of fluorescent-labeled clathrin (protein involved in endocytosis via clathrin pathway), expressed in transfected 3T3 fibroblasts cells. Using a model polymeric colloid and cells with a fluorescently tagged clathrin endocytosis pathway, we demonstrate that wide field TIRM/TIRF coimaging enables live visualization of the process of colloidal particle interaction with the labeled cell structure, which is valuable for discerning the membrane events and route of colloid internalization by the cell. We further show that 500 nm in diameter model polystyrene colloid associates with clathrin, prior to and during its cellular internalization. This association is not apparent with larger, 1 μm in diameter colloids, indicating an upper particle size limit for clathrin-mediated endocytosis.
Johnson, Heath E; Haugh, Jason M
2013-12-02
This unit focuses on the use of total internal reflection fluorescence (TIRF) microscopy and image analysis methods to study the dynamics of signal transduction mediated by class I phosphoinositide 3-kinases (PI3Ks) in mammalian cells. The first four protocols cover live-cell imaging experiments, image acquisition parameters, and basic image processing and segmentation. These methods are generally applicable to live-cell TIRF experiments. The remaining protocols outline more advanced image analysis methods, which were developed in our laboratory for the purpose of characterizing the spatiotemporal dynamics of PI3K signaling. These methods may be extended to analyze other cellular processes monitored using fluorescent biosensors. Copyright © 2013 John Wiley & Sons, Inc.
Hageneder, Simone; Bauch, Martin; Dostalek, Jakub
2016-08-15
This paper investigates plasmonic amplification in two commonly used optical configurations for fluorescence readout of bioassays - epifluorescence (EPF) and total internal reflection fluorescence (TIRF). The plasmonic amplification in the EPF configuration was implemented by using crossed gold diffraction grating and Kretschmann geometry of attenuated total reflection method (ATR) was employed in the TIRF configuration. Identical assay, surface architecture for analyte capture, and optics for the excitation, collection and detection of emitted fluorescence light intensity were used in both TIRF and EPF configurations. Simulations predict that the crossed gold diffraction grating (EPF) can amplify the fluorescence signal by a factor of 10(2) by the combination of surface plasmon-enhanced excitation and directional surface plasmon-coupled emission in the red part of spectrum. This factor is about order of magnitude higher than that predicted for the Kretschmann geometry (TIRF) which only took advantage of the surface plasmon-enhanced excitation. When applied for the readout of sandwich interleukin 6 (IL-6) immunoassay, the plasmonically amplified EPF geometry designed for Alexa Fluor 647 labels offered 4-times higher fluorescence signal intensity compared to TIRF. Interestingly, both geometries allowed reaching the same detection limit of 0.4pM despite of the difference in the fluorescence signal enhancement. This is attributed to inherently lower background of fluorescence signal for TIRF geometry compared to that for EPF which compensates for the weaker fluorescence signal enhancement. The analysis of the inflammation biomarker IL-6 in serum at medically relevant concentrations and the utilization of plasmonic amplification for the fluorescence measurement of kinetics of surface affinity reactions are demonstrated for both EPF and TIRF readout. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.
Three-dimensional super-resolved live cell imaging through polarized multi-angle TIRF.
Zheng, Cheng; Zhao, Guangyuan; Liu, Wenjie; Chen, Youhua; Zhang, Zhimin; Jin, Luhong; Xu, Yingke; Kuang, Cuifang; Liu, Xu
2018-04-01
Measuring three-dimensional nanoscale cellular structures is challenging, especially when the structure is dynamic. Owing to the informative total internal reflection fluorescence (TIRF) imaging under varied illumination angles, multi-angle (MA) TIRF has been examined to offer a nanoscale axial and a subsecond temporal resolution. However, conventional MA-TIRF still performs badly in lateral resolution and fails to characterize the depth image in densely distributed regions. Here, we emphasize the lateral super-resolution in the MA-TIRF, exampled by simply introducing polarization modulation into the illumination procedure. Equipped with a sparsity and accelerated proximal algorithm, we examine a more precise 3D sample structure compared with previous methods, enabling live cell imaging with a temporal resolution of 2 s and recovering high-resolution mitochondria fission and fusion processes. We also shared the recovery program, which is the first open-source recovery code for MA-TIRF, to the best of our knowledge.
NASA Astrophysics Data System (ADS)
Cardoso Dos Santos, Marcelina; Vézy, Cyrille; Jaffiol, Rodolphe
2016-02-01
Total Internal Reflection Fluorescence Microscopy (TIRFM) is a widespread technique to study cellular process occurring near the contact region with the glass substrate. In this field, determination of the accurate distance from the surface to the plasma membrane constitutes a crucial issue to investigate the physical basis of cellular adhesion process. However, quantitative interpretation of TIRF pictures regarding the distance z between a labeled membrane and the substrate is not trivial. Indeed, the contrast of TIRF images depends on several parameters more and less well known (local concentration of dyes, absorption cross section, angular emission pattern…). The strategy to get around this problem is to exploit a series of TIRF pictures recorded at different incident angles in evanescent regime. This technique called variable-angle TIRF microscopy (vaTIRFM), allowing to map the membrane-substrate separation distance with a nanometric resolution (10-20 nm). vaTIRFM was developed by Burmeister, Truskey and Reichert in the early 1990s with a prism-based TIRF setup [Journal of Microscopy 173, 39-51 (1994)]. We propose a more convenient prismless setup, which uses only a rotatable mirror to adjust precisely the laser beam on the back focal plane of the oil immersion objective (no azimuthal scanning is needed). The series of TIRF images permit us to calculate accurately membrane-surface distances in each pixel. We demonstrate that vaTIRFM are useful to quantify the adhesion of living cells for specific and unspecific membrane-surface interactions, achieved on various functionalized substrates with polymers (BSA, poly-L-lysin) or extracellular matrix proteins (collagen and fibronectin).
Simmert, Steve; Abdosamadi, Mohammad Kazem; Hermsdorf, Gero; Schäffer, Erik
2018-05-28
Optical tweezers combined with various microscopy techniques are a versatile tool for single-molecule force spectroscopy. However, some combinations may compromise measurements. Here, we combined optical tweezers with total-internal-reflection-fluorescence (TIRF) and interference-reflection microscopy (IRM). Using a light-emitting diode (LED) for IRM illumination, we show that single microtubules can be imaged with high contrast. Furthermore, we converted the IRM interference pattern of an upward bent microtubule to its three-dimensional (3D) profile calibrated against the optical tweezers and evanescent TIRF field. In general, LED-based IRM is a powerful method for high-contrast 3D microscopy.
Zobiak, Bernd; Failla, Antonio Virgilio
2018-03-01
Understanding the cellular processes that occur between the cytosol and the plasma membrane is an important task for biological research. Till now, however, it was not possible to combine fast and high-resolution imaging of both the isolated plasma membrane and the surrounding intracellular volume. Here, we demonstrate the combination of fast high-resolution spinning disk (SD) and total internal reflection fluorescence (TIRF) microscopy for specific imaging of the plasma membrane. A customised SD-TIRF microscope was used with specific design of the light paths that allowed, for the first time, live SD-TIRF experiments at high acquisition rates. A series of experiments is shown to demonstrate the feasibility and performance of our setup. © 2017 The Authors. Journal of Microscopy published by JohnWiley & Sons Ltd on behalf of Royal Microscopical Society.
Compact 3D printed module for fluorescence and label-free imaging using evanescent excitation
NASA Astrophysics Data System (ADS)
Pandey, Vikas; Gupta, Shalini; Elangovan, Ravikrishnan
2018-01-01
Total internal reflection fluorescence (TIRF) microscopy is widely used for selective excitation and high-resolution imaging of fluorophores, and more recently label-free nanosized objects, with high vertical confinement near a liquid-solid interface. Traditionally, high numerical aperture objectives (>1.4) are used to simultaneously generate evanescent waves and collect fluorescence emission signals which limits their use to small area imaging (<0.1 mm2). Objective-based TIRFs are also expensive as they require dichroic mirrors and efficient notch filters to prevent specular reflection within the objective lenses. We have developed a compact 3D module called cTIRF that can generate evanescent waves in microscope glass slides via a planar waveguide illumination. The module can be attached as a fixture to any existing optical microscope, converting it into a TIRF and enabling high signal-to-noise ratio (SNR) fluorescence imaging using any magnification objective. As the incidence optics is perpendicular to the detector, label-free evanescent scattering-based imaging of submicron objects can also be performed without using emission filters. SNR is significantly enhanced in this case as compared to cTIRF alone, as seen through our model experiments performed on latex beads and mammalian cells. Extreme flexibility and the low cost of our approach makes it scalable for limited resource settings.
Topography of Cells Revealed by Variable-Angle Total Internal Reflection Fluorescence Microscopy.
Cardoso Dos Santos, Marcelina; Déturche, Régis; Vézy, Cyrille; Jaffiol, Rodolphe
2016-09-20
We propose an improved version of variable-angle total internal reflection fluorescence microscopy (vaTIRFM) adapted to modern TIRF setup. This technique involves the recording of a stack of TIRF images, by gradually increasing the incident angle of the light beam on the sample. A comprehensive theory was developed to extract the membrane/substrate separation distance from fluorescently labeled cell membranes. A straightforward image processing was then established to compute the topography of cells with a nanometric axial resolution, typically 10-20 nm. To highlight the new opportunities offered by vaTIRFM to quantify adhesion process of motile cells, adhesion of MDA-MB-231 cancer cells on glass substrate coated with fibronectin was examined. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.
NASA Astrophysics Data System (ADS)
Zude, Manuela; Spinelli, Lorenzo; Dosche, Carsten; Torricelli, Alessandro
2009-08-01
In sweet cherry (Prunus avium), the red pigmentation is correlated with the fruit maturity stage and can be measured by non-invasive spectroscopy. In the present study, the influence of varying fruit scattering coefficients on the fruit remittance spectrum (cw) were corrected with the effective pathlength and refractive index in the fruit tissue obtained with distribution of time-of-flight (DTOF) readings and total internal reflection fluorescence (TIRF) analysis, respectively. The approach was validated on fruits providing variation in the scattering coefficient outside the calibration sample set. In the validation, the measuring uncertainty when non-invasively analyzing fruits with cw method in comparison with combined application of cw, DTOF, and TIRF measurements showed an increase in r2 up to 22.7 % with, however, high errors in all approaches.
Devauges, Viviane; Matthews, Daniel R.; Aluko, Justin; Nedbal, Jakub; Levitt, James A.; Poland, Simon P.; Coban, Oana; Weitsman, Gregory; Monypenny, James; Ng, Tony; Ameer-Beg, Simon M.
2014-01-01
We present a novel imaging system combining total internal reflection fluorescence (TIRF) microscopy with measurement of steady-state acceptor fluorescence anisotropy in order to perform live cell Förster Resonance Energy Transfer (FRET) imaging at the plasma membrane. We compare directly the imaging performance of fluorescence anisotropy resolved TIRF with epifluorescence illumination. The use of high numerical aperture objective for TIRF required correction for induced depolarization factors. This arrangement enabled visualisation of conformational changes of a Raichu-Cdc42 FRET biosensor by measurement of intramolecular FRET between eGFP and mRFP1. Higher activity of the probe was found at the cell plasma membrane compared to intracellularly. Imaging fluorescence anisotropy in TIRF allowed clear differentiation of the Raichu-Cdc42 biosensor from negative control mutants. Finally, inhibition of Cdc42 was imaged dynamically in live cells, where we show temporal changes of the activity of the Raichu-Cdc42 biosensor. PMID:25360776
Brunstein, Maia; Hérault, Karine; Oheim, Martin
2014-01-01
Azimuthal beam scanning makes evanescent-wave (EW) excitation isotropic, thereby producing total internal reflection fluorescence (TIRF) images that are evenly lit. However, beam spinning does not fundamentally address the problem of propagating excitation light that is contaminating objective-type TIRF. Far-field excitation depends more on the specific objective than on cell scattering. As a consequence, the excitation impurities in objective-type TIRF are only weakly affected by changes of azimuthal or polar beam angle. These are the main results of the first part of this study (Eliminating unwanted far-field excitation in objective-type TIRF. Pt.1. Identifying sources of nonevanescent excitation light). This second part focuses on exactly where up beam in the illumination system stray light is generated that gives rise to nonevanescent components in TIRF. Using dark-field imaging of scattered excitation light we pinpoint the objective, intermediate lenses and, particularly, the beam scanner as the major sources of stray excitation. We study how adhesion-molecule coating and astrocytes or BON cells grown on the coverslip surface modify the dark-field signal. On flat and weakly scattering cells, most background comes from stray reflections produced far from the sample plane, in the beam scanner and the objective lens. On thick, optically dense cells roughly half of the scatter is generated by the sample itself. We finally show that combining objective-type EW excitation with supercritical-angle fluorescence (SAF) detection efficiently rejects the fluorescence originating from deeper sample regions. We demonstrate that SAF improves the surface selectivity of TIRF, even at shallow penetration depths. The coplanar microscopy scheme presented here merges the benefits of beam spinning EW excitation and SAF detection and provides the conditions for quantitative wide-field imaging of fluorophore dynamics at or near the plasma membrane. PMID:24606929
Near-Membrane Refractometry Using Supercritical Angle Fluorescence.
Brunstein, Maia; Roy, Lopamudra; Oheim, Martin
2017-05-09
Total internal reflection fluorescence (TIRF) microscopy and its variants are key technologies for visualizing the dynamics of single molecules or organelles in live cells. Yet truly quantitative TIRF remains problematic. One unknown hampering the interpretation of evanescent-wave excited fluorescence intensities is the undetermined cell refractive index (RI). Here, we use a combination of TIRF excitation and supercritical angle fluorescence emission detection to directly measure the average RI in the "footprint" region of the cell during image acquisition. Our RI measurement is based on the determination on a back-focal plane image of the critical angle separating evanescent and far-field fluorescence emission components. We validate our method by imaging mouse embryonic fibroblasts and BON cells. By targeting various dyes and fluorescent-protein chimeras to vesicles, the plasma membrane, as well as mitochondria and the endoplasmic reticulum, we demonstrate local RI measurements with subcellular resolution on a standard TIRF microscope, with a removable Bertrand lens as the only modification. Our technique has important applications for imaging axial vesicle dynamics and the mitochondrial energy state or detecting metabolically more active cancer cells. Copyright © 2017. Published by Elsevier Inc.
Cardoso Dos Santos, Marcelina; Vézy, Cyrille; Jaffiol, Rodolphe
2016-06-01
We recently proposed a straightforward fluorescence microscopy technique to study adhesion of Giant Unilamellar Vesicles. This technique is based on dual observations which combine epi-fluorescence microscopy and total internal reflection fluorescence (TIRF) microscopy: TIRF images are normalized by epi-fluorescence ones. By this way, it is possible to map the membrane/substrate separation distance with a nanometric resolution, typically ~20 nm, with a maximal working range of 300-400 nm. The purpose of this paper is to demonstrate that this technique is useful to quantify vesicle adhesion from ultra-weak to strong membrane-surface interactions. Thus, we have examined unspecific and specific adhesion conditions. Concerning unspecific adhesion, we have controlled the strength of electrostatic forces between negatively charged vesicles and various functionalized surfaces which exhibit a positive or a negative effective charge. Specific adhesion was highlighted with lock-and-key forces mediated by the well defined biotin/streptavidin recognition. Copyright © 2016 Elsevier B.V. All rights reserved.
Wang, Jun; Fei, Bei; Geahlen, Robert L.
2010-01-01
Protein translocation, or the change in a protein’s location between different subcellular compartments, is a critical process by which intracellular proteins carry out their cellular functions. Aberrant translocation events contribute to various diseases ranging from metabolic disorders to cancer. In this study, we demonstrate the use of a newly developed single-cell tool, microfluidic total internal reflection fluorescence flow cytometry (TIRF-FC), for detecting both cytosol to plasma membrane and cytosol to nucleus translocations using the tyrosine kinase Syk and the transcription factor NF-κB as models. This technique detects fluorescent molecules at the plasma membrane and in the membrane-proximal cytosol in single cells. We were able to record quantitatively changes in the fluorescence density in the evanescent field associated with these translocation processes for large cell populations with single cell resolution. We envision that TIRF-FC will provide a new approach to explore the molecular biology and clinical relevance of protein translocations. PMID:20820633
Larson, Joshua; Kirk, Matt; Drier, Eric A.; O’Brien, William; MacKay, James F.; Friedman, Larry; Hoskins, Aaron
2015-01-01
Colocalization Single Molecule Spectroscopy (CoSMoS) has proven to be a useful method for studying the composition, kinetics, and mechanisms of complex cellular machines. Key to the technique is the ability to simultaneously monitor multiple proteins and/or nucleic acids as they interact with one another. Here we describe a protocol for constructing a CoSMoS micromirror Total Internal Reflection Fluorescence Microscope (mmTIRFM). Design and construction of a scientific microscope often requires a number of custom components and a significant time commitment. In our protocol, we have streamlined this process by implementation of a commercially available microscopy platform designed to accommodate the optical components necessary for a mmTIRFM. The mmTIRF system eliminates the need for machining custom parts by the end-user and facilitates optical alignment. Depending on the experience-level of the microscope builder, these time-savings and the following protocol can enable mmTIRF construction to be completed within two months. PMID:25188633
Larson, Joshua; Kirk, Matt; Drier, Eric A; O'Brien, William; MacKay, James F; Friedman, Larry J; Hoskins, Aaron A
2014-10-01
Colocalization single-molecule spectroscopy (CoSMoS) has proven to be a useful method for studying the composition, kinetics and mechanisms of complex cellular machines. Key to the technique is the ability to simultaneously monitor multiple proteins and/or nucleic acids as they interact with one another. Here we describe a protocol for constructing a CoSMoS micromirror total internal reflection fluorescence microscope (mmTIRFM). Design and construction of a scientific microscope often requires a number of custom components and a substantial time commitment. In our protocol, we have streamlined this process by implementation of a commercially available microscopy platform designed to accommodate the optical components necessary for an mmTIRFM. The mmTIRF system eliminates the need for machining custom parts by the end user and facilitates optical alignment. Depending on the experience level of the microscope builder, these time savings and the following protocol can enable mmTIRF construction to be completed within 2 months.
Brunstein, Maia; Teremetz, Maxime; Hérault, Karine; Tourain, Christophe; Oheim, Martin
2014-01-01
Total internal reflection fluorescence microscopy (TIRFM) achieves subdiffraction axial sectioning by confining fluorophore excitation to a thin layer close to the cell/substrate boundary. However, it is often unknown how thin this light sheet actually is. Particularly in objective-type TIRFM, large deviations from the exponential intensity decay expected for pure evanescence have been reported. Nonevanescent excitation light diminishes the optical sectioning effect, reduces contrast, and renders TIRFM-image quantification uncertain. To identify the sources of this unwanted fluorescence excitation in deeper sample layers, we here combine azimuthal and polar beam scanning (spinning TIRF), atomic force microscopy, and wavefront analysis of beams passing through the objective periphery. Using a variety of intracellular fluorescent labels as well as negative staining experiments to measure cell-induced scattering, we find that azimuthal beam spinning produces TIRFM images that more accurately portray the real fluorophore distribution, but these images are still hampered by far-field excitation. Furthermore, although clearly measureable, cell-induced scattering is not the dominant source of far-field excitation light in objective-type TIRF, at least for most types of weakly scattering cells. It is the microscope illumination optical path that produces a large cell- and beam-angle invariant stray excitation that is insensitive to beam scanning. This instrument-induced glare is produced far from the sample plane, inside the microscope illumination optical path. We identify stray reflections and high-numerical aperture aberrations of the TIRF objective as one important source. This work is accompanied by a companion paper (Pt.2/2). PMID:24606927
Wang, M D; Axelrod, D
1994-09-01
To study when and where acetylcholine receptor (AChR) clusters appear on developing rat myotubes in primary culture, we have made time-lapse movies of total internal reflection fluorescence (TIRF) overlaid with schlieren transmitted light images. The receptors, including the ones newly incorporated into the membrane, were labeled with rhodamine alpha-bungarotoxin (R-BT) continuously present in the medium. Since TIRF illuminates only cell-substrate contact regions where almost all of the AChR clusters are located, background fluorescence from fluorophores either in the bulk solution or inside the cells can be suppressed. Also, because TIRF minimizes the exposure of the cell interior to light, the healthy survival of the culture during imaging procedures is much enhanced relative to standard epi- (or trans-) illumination. During the experiment, cells were kept alive on the microscope stage at 37 degrees C in an atmosphere of 10% CO2. Two digital images were recorded by a CCD camera every 20 min: the schlieren image of the cells and the TIRF image of the clusters. After background subtraction, the cluster image was displayed in pseudocolors, overlaid onto the cell images, and recorded as 3 frames on a videotape. The final movies are thus able to summarize a week-long experiment in less than a minute. These movies and images show that clusters form often shortly after the myoblast fusion but sometimes much later, and the formation takes place very rapidly (a few hours). The clusters have an average lifetime of around a day, much shorter than the lifetime of a typical myotube. The brightest and largest clusters tend to be the longest-lived. The cluster formation seems to be associated with the contacts of myotubes at the glass substrate, but not with cell-cell contacts or myoblast fusion into myotubes. New AChR continuously appear in preexisting clusters: after photobleaching, the fluorescence of some clusters recovers within an hour.
A programmable light engine for quantitative single molecule TIRF and HILO imaging.
van 't Hoff, Marcel; de Sars, Vincent; Oheim, Martin
2008-10-27
We report on a simple yet powerful implementation of objective-type total internal reflection fluorescence (TIRF) and highly inclined and laminated optical sheet (HILO, a type of dark-field) illumination. Instead of focusing the illuminating laser beam to a single spot close to the edge of the microscope objective, we are scanning during the acquisition of a fluorescence image the focused spot in a circular orbit, thereby illuminating the sample from various directions. We measure parameters relevant for quantitative image analysis during fluorescence image acquisition by capturing an image of the excitation light distribution in an equivalent objective backfocal plane (BFP). Operating at scan rates above 1 MHz, our programmable light engine allows directional averaging by circular spinning the spot even for sub-millisecond exposure times. We show that restoring the symmetry of TIRF/HILO illumination reduces scattering and produces an evenly lit field-of-view that affords on-line analysis of evanescnt-field excited fluorescence without pre-processing. Utilizing crossed acousto-optical deflectors, our device generates arbitrary intensity profiles in BFP, permitting variable-angle, multi-color illumination, or objective lenses to be rapidly exchanged.
Optimal Background Estimators in Single-Molecule FRET Microscopy.
Preus, Søren; Hildebrandt, Lasse L; Birkedal, Victoria
2016-09-20
Single-molecule total internal reflection fluorescence (TIRF) microscopy constitutes an umbrella of powerful tools that facilitate direct observation of the biophysical properties, population heterogeneities, and interactions of single biomolecules without the need for ensemble synchronization. Due to the low signal/noise ratio in single-molecule TIRF microscopy experiments, it is important to determine the local background intensity, especially when the fluorescence intensity of the molecule is used quantitatively. Here we compare and evaluate the performance of different aperture-based background estimators used particularly in single-molecule Förster resonance energy transfer. We introduce the general concept of multiaperture signatures and use this technique to demonstrate how the choice of background can affect the measured fluorescence signal considerably. A new, to our knowledge, and simple background estimator is proposed, called the local statistical percentile (LSP). We show that the LSP background estimator performs as well as current background estimators at low molecular densities and significantly better in regions of high molecular densities. The LSP background estimator is thus suited for single-particle TIRF microscopy of dense biological samples in which the intensity itself is an observable of the technique. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Force determination in lateral magnetic tweezers combined with TIRF microscopy.
Madariaga-Marcos, J; Hormeño, S; Pastrana, C L; Fisher, G L M; Dillingham, M S; Moreno-Herrero, F
2018-03-01
Combining single-molecule techniques with fluorescence microscopy has attracted much interest because it allows the correlation of mechanical measurements with directly visualized DNA : protein interactions. In particular, its combination with total internal reflection fluorescence microscopy (TIRF) is advantageous because of the high signal-to-noise ratio this technique achieves. This, however, requires stretching long DNA molecules across the surface of a flow cell to maximize polymer exposure to the excitation light. In this work, we develop a module to laterally stretch DNA molecules at a constant force, which can be easily implemented in regular or combined magnetic tweezers (MT)-TIRF setups. The pulling module is further characterized in standard flow cells of different thicknesses and glass capillaries, using two types of micrometer size superparamagnetic beads, long DNA molecules, and a home-built device to rotate capillaries with mrad precision. The force range achieved by the magnetic pulling module was between 0.1 and 30 pN. A formalism for estimating forces in flow-stretched tethered beads is also proposed, and the results compared with those of lateral MT, demonstrating that lateral MT achieve higher forces with lower dispersion. Finally, we show the compatibility with TIRF microscopy and the parallelization of measurements by characterizing DNA binding by the centromere-binding protein ParB from Bacillus subtilis. Simultaneous MT pulling and fluorescence imaging demonstrate the non-specific binding of BsParB on DNA under conditions restrictive to condensation.
Total-Internal-Reflection Platforms for Chemical and Biological Sensing Applications
NASA Astrophysics Data System (ADS)
Sapsford, Kim E.
Sensing platforms based on the principle of total internal reflection (TIR) represent a fairly mature yet still expanding and exciting field of research. Sensor development has mainly been driven by the need for rapid, stand-alone, automated devices for application in the fields of clinical diagnosis and screening, food and water safety, environmental monitoring, and chemical and biological warfare agent detection. The technologies highlighted in this chapter are continually evolving, taking advantage of emerging advances in microfabrication, lab-on-a-chip, excitation, and detection techniques. This chapter describes many of the underlying principles of TIR-based sensing platforms and additionally focusses on planar TIR fluorescence (TIRF)-based chemical and biological sensors.
Real-time TIRF observation of vinculin recruitment to stretched α-catenin by AFM.
Maki, Koichiro; Han, Sung-Woong; Hirano, Yoshinori; Yonemura, Shigenobu; Hakoshima, Toshio; Adachi, Taiji
2018-01-25
Adherens junctions (AJs) adaptively change their intensities in response to intercellular tension; therefore, they integrate tension generated by individual cells to drive multicellular dynamics, such as morphogenetic change in embryos. Under intercellular tension, α-catenin, which is a component protein of AJs, acts as a mechano-chemical transducer to recruit vinculin to promote actin remodeling. Although in vivo and in vitro studies have suggested that α-catenin-mediated mechanotransduction is a dynamic molecular process, which involves a conformational change of α-catenin under tension to expose a cryptic vinculin binding site, there are no suitable experimental methods to directly explore the process. Therefore, in this study, we developed a novel system by combining atomic force microscopy (AFM) and total internal reflection fluorescence (TIRF). In this system, α-catenin molecules (residues 276-634; the mechano-sensitive M 1 -M 3 domain), modified on coverslips, were stretched by AFM and their recruitment of Alexa-labeled full-length vinculin molecules, dissolved in solution, were observed simultaneously, in real time, using TIRF. We applied a physiologically possible range of tensions and extensions to α-catenin and directly observed its vinculin recruitment. Our new system could be used in the fields of mechanobiology and biophysics to explore functions of proteins under tension by coupling biomechanical and biochemical information.
Single Event Resolution of Plant Plasma Membrane Protein Endocytosis by TIRF Microscopy.
Johnson, Alexander; Vert, Grégory
2017-01-01
Endocytosis is a key process in the internalization of extracellular materials and plasma membrane proteins, such as receptors and transporters, thereby controlling many aspects of cell signaling and cellular homeostasis. Endocytosis in plants has an essential role not only for basic cellular functions but also for growth and development, nutrient delivery, toxin avoidance, and pathogen defense. The precise mechanisms of endocytosis in plants remain quite elusive. The lack of direct visualization and examination of single events of endocytosis has greatly hampered our ability to precisely monitor the cell surface lifetime and the recruitment profile of proteins driving endocytosis or endocytosed cargos in plants. Here, we discuss the necessity to systematically implement total internal reflection fluorescence microcopy (TIRF) in the Plant Cell Biology community and present reliable protocols for high spatial and temporal imaging of endocytosis in plants using clathrin-mediated endocytosis as a test case, since it represents the major route for internalization of cell-surface proteins in plants. We developed a robust method to directly visualize cell surface proteins using TIRF microscopy combined to a high throughput, automated and unbiased analysis pipeline to determine the temporal recruitment profile of proteins to single sites of endocytosis, using the departure of clathrin as a physiological reference for scission. Using this 'departure assay', we assessed the recruitment of two different AP-2 subunits, alpha and mu, to the sites of endocytosis and found that AP2A1 was recruited in concert with clathrin, while AP2M was not. This validated approach therefore offers a powerful solution to better characterize the plant endocytic machinery and the dynamics of one's favorite cargo protein.
NASA Astrophysics Data System (ADS)
Ströhl, Florian; Wong, Hovy H. W.; Holt, Christine E.; Kaminski, Clemens F.
2018-01-01
Fluorescence anisotropy imaging microscopy (FAIM) measures the depolarization properties of fluorophores to deduce molecular changes in their environment. For successful FAIM, several design principles have to be considered and a thorough system-specific calibration protocol is paramount. One important calibration parameter is the G factor, which describes the system-induced errors for different polarization states of light. The determination and calibration of the G factor is discussed in detail in this article. We present a novel measurement strategy, which is particularly suitable for FAIM with high numerical aperture objectives operating in TIRF illumination mode. The method makes use of evanescent fields that excite the sample with a polarization direction perpendicular to the image plane. Furthermore, we have developed an ImageJ/Fiji plugin, AniCalc, for FAIM data processing. We demonstrate the capabilities of our TIRF-FAIM system by measuring β -actin polymerization in human embryonic kidney cells and in retinal neurons.
Slow fusion pore expansion creates a unique reaction chamber for co-packaged cargo
Bittner, Mary A.; Lawrence, Daniel A.
2017-01-01
A lumenal secretory granule protein, tissue plasminogen activator (tPA), greatly slows fusion pore dilation and thereby slows its own discharge. We investigated another outcome of the long-lived narrow fusion pore: the creation of a nanoscale chemical reaction chamber for granule contents in which the pH is suddenly neutralized upon fusion. Bovine adrenal chromaffin cells endogenously express both tPA and its primary protein inhibitor, plasminogen activator inhibitor 1 (PAI). We found by immunocytochemistry that tPA and PAI are co-packaged in the same secretory granule. It is known that PAI irreversibly and covalently inactivates tPA at neutral pH. We demonstrate with zymography that the acidic granule lumen protects tPA from inactivation by PAI. Immunocytochemistry, total internal reflection fluorescence (TIRF) microscopy, and polarized TIRF microscopy demonstrated that co-packaged PAI and tPA remain together in granules for many seconds in the nanoscale reaction chamber, more than enough time to inhibit tPA and create a new secreted protein species. PMID:28882880
Resolving dynamics of cell signaling via real-time imaging of the immunological synapse.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Stevens, Mark A.; Pfeiffer, Janet R.; Wilson, Bridget S.
2009-10-01
This highly interdisciplinary team has developed dual-color, total internal reflection microscopy (TIRF-M) methods that enable us to optically detect and track in real time protein migration and clustering at membrane interfaces. By coupling TIRF-M with advanced analysis techniques (image correlation spectroscopy, single particle tracking) we have captured subtle changes in membrane organization that characterize immune responses. We have used this approach to elucidate the initial stages of cell activation in the IgE signaling network of mast cells and the Toll-like receptor (TLR-4) response in macrophages stimulated by bacteria. To help interpret these measurements, we have undertaken a computational modeling effortmore » to connect the protein motion and lipid interactions. This work provides a deeper understanding of the initial stages of cellular response to external agents, including dynamics of interaction of key components in the signaling network at the 'immunological synapse,' the contact region of the cell and its adversary.« less
Optical biosensor system with integrated microfluidic sample preparation and TIRF based detection
NASA Astrophysics Data System (ADS)
Gilli, Eduard; Scheicher, Sylvia R.; Suppan, Michael; Pichler, Heinz; Rumpler, Markus; Satzinger, Valentin; Palfinger, Christian; Reil, Frank; Hajnsek, Martin; Köstler, Stefan
2013-05-01
There is a steadily growing demand for miniaturized bioanalytical devices allowing for on-site or point-of-care detection of biomolecules or pathogens in applications like diagnostics, food testing, or environmental monitoring. These, so called labs-on-a-chip or micro-total analysis systems (μ-TAS) should ideally enable convenient sample-in - result-out type operation. Therefore, the entire process from sample preparation, metering, reagent incubation, etc. to detection should be performed on a single disposable device (on-chip). In the early days such devices were mainly fabricated using glass or silicon substrates and adapting established fabrication technologies from the electronics and semiconductor industry. More recently, the development focuses on the use of thermoplastic polymers as they allow for low-cost high volume fabrication of disposables. One of the most promising materials for the development of plastic based lab-on-achip systems are cyclic olefin polymers and copolymers (COP/COC) due to their excellent optical properties (high transparency and low autofluorescence) and ease of processing. We present a bioanalytical system for whole blood samples comprising a disposable plastic chip based on TIRF (total internal reflection fluorescence) optical detection. The chips were fabricated by compression moulding of COP and microfluidic channels were structured by hot embossing. These microfluidic structures integrate several sample pretreatment steps. These are the separation of erythrocytes, metering of sample volume using passive valves, and reagent incubation for competitive bioassays. The surface of the following optical detection zone is functionalized with specific capture probes in an array format. The plastic chips comprise dedicated structures for simple and effective coupling of excitation light from low-cost laser diodes. This enables TIRF excitation of fluorescently labeled probes selectively bound to detection spots at the microchannel surface. The fluorescence of these detection arrays is imaged using a simple set-up based on a digital consumer camera. Image processing for spot detection and intensity calculation is accomplished using customized software. Using this combined TIRF excitation and imaging based detection approach allowes for effective suppression of background fluorescence from the sample, multiplexed detection in an array format, as well as internal calibration and background correction.
Surface plasmon-assisted microscope.
Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal; Joshi, Chaitanya R; Borgmann, Kathleen; Ghorpade, Anuja; Gryczynski, Ignacy
2018-06-01
Total internal reflection microscopy (TIRF) has been a powerful tool in biological research. The most valuable feature of the method has been the ability to image 100- to 200-nm-thick layer of cell features adjacent to a coverslip, such as membrane lipids, membrane receptors, and structures proximal-to-basal membranes. Here, we demonstrate an alternative method of imaging thin-layer proximal-to-basal membranes by placing a sample on a high refractive index coverslip covered by a thin layer of gold. The sample is illuminated using the Kretschmann method (i.e., from the top to an aqueous medium). Fluorophores that are close to the metal surface induce surface plasmons in the metal film. Fluorescence from fluorophores near the metal surface couple with surface plasmons allowing them to penetrate the metal surface and emerge at a surface plasmon coupled emission angle. The thickness of the detection layer is further reduced in comparison with TIRF by metal quenching of fluorophores at a close proximity (below 10 nm) to a surface. Fluorescence is collected by a high NA objective and imaged by EMCCD or converted to a signal by avalanche photodiode fed by a single-mode optical fiber inserted in the conjugate image plane of the objective. The system avoids complications of through-the-objective TIRF associated with shared excitation and emission light path, has thin collection thickness, produces excellent background rejection, and is an effective method to study molecular motion. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ohara-Imaizumi, Mica; Aoyagi, Kyota; Akimoto, Yoshihiro
To analyze the exocytosis of glucagon-like peptide-1 (GLP-1) granules, we imaged the motion of GLP-1 granules labeled with enhanced yellow fluorescent protein (Venus) fused to human growth hormone (hGH-Venus) in an enteroendocrine cell line, STC-1 cells, by total internal reflection fluorescent (TIRF) microscopy. We found glucose stimulation caused biphasic GLP-1 granule exocytosis: during the first phase, fusion events occurred from two types of granules (previously docked granules and newcomers), and thereafter continuous fusion was observed mostly from newcomers during the second phase. Closely similar to the insulin granule fusion from pancreatic {beta} cells, the regulated biphasic exocytosis from two typesmore » of granules may be a common mechanism in glucose-evoked hormone release from endocrine cells.« less
Buijs; Hlady
1997-06-01
Interactions of recombinant human growth hormone and lysozyme with solid surfaces are studied using total internal reflection fluorescence (TIRF) and monitoring the protein's intrinsic tryptophan fluorescence. The intensity, spectra, quenching, and polarization of the fluorescence emitted by the adsorbed proteins are monitored and related to adsorption kinetics, protein conformation, and fluorophore rotational mobility. To study the influence of electrostatic and hydrophobic interactions on the adsorption process, three sorbent surfaces are used which differ in charge and hydrophobicity. The chemical surface groups are silanol, methyl, and quaternary amine. Results indicate that adsorption of hGH is dominated by hydrophobic interactions. Lysozyme adsoption is strongly affected by the ionic strength. This effect is probably caused by an ionic strength dependent conformational state in solution which, in turn, influences the affinity for adsorption. Both proteins are more strongly bound to hydrophobic surfaces and this strong interaction is accompanied by a less compact conformation. Furthermore, it was seen that regardless of the characteristics of the sorbent surface, the rotational mobility of both proteins' tryptophans is largely reduced upon adsorption.
Fluorescent diamond nanoparticle as a probe of intracellular traffic in primary neurons in culture
NASA Astrophysics Data System (ADS)
Le, Xuan Loc; Lepagnol-Bestel, Aude-Marie; Adam, Marie-Pierre; Thomas, Alice; Dantelle, Géraldine; Chang, Cheng-Chun; Mohan, Nitin; Chang, Huan-Cheng; Treussart, François; Simonneau, Michel
2012-03-01
Neurons display dendritic spines plasticity and morphology anomalies in numerous psychiatric and neurodegenerative diseases. These changes are associated to abnormal dendritic traffic that can be evidenced by fluorescence microscopy. As a fluorescent probe we propose to use fluorescent diamond nanoparticles with size of < 50 nm. Color centers embedded inside the diamond nanoparticles are perfectly photostable emitters allowing for long-term tracking. Nanodiamond carbon surface is also well suited for biomolecule functionalization to target specific cellular compartments. We show that fluorescent nanodiamonds can be spontaneously internalized in neurons in culture and imaged by confocal and Total Internal Reflection (TIRF) microscopy with a high signal over background ratio.
Label free imaging of cell-substrate contacts by holographic total internal reflection microscopy.
Mandracchia, Biagio; Gennari, Oriella; Marchesano, Valentina; Paturzo, Melania; Ferraro, Pietro
2017-09-01
The study of cell adhesion contacts is pivotal to understand cell mechanics and interaction at substrates or chemical and physical stimuli. We designed and built a HoloTIR microscope for label-free quantitative phase imaging of total internal reflection. Here we show for the first time that HoloTIR is a good choice for label-free study of focal contacts and of cell/substrate interaction as its sensitivity is enhanced in comparison with standard TIR microscopy. Finally, the simplicity of implementation and relative low cost, due to the requirement of less optical components, make HoloTIR a reasonable alternative, or even an addition, to TIRF microscopy for mapping cell/substratum topography. As a proof of concept, we studied the formation of focal contacts of fibroblasts on three substrates with different levels of affinity for cell adhesion. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
Zhang, Hui; Guo, Peixuan
2014-05-15
Direct counting of biomolecules within biological complexes or nanomachines is demanding. Single molecule counting using optical microscopy is challenging due to the diffraction limit. The single molecule photobleaching (SMPB) technology for direct counting developed by our team (Shu et al., 2007 [18]; Zhang et al., 2007 [19]) offers a simple and straightforward method to determine the stoichiometry of molecules or subunits within biocomplexes or nanomachines at nanometer scales. Stoichiometry is determined by real-time observation of the number of descending steps resulted from the photobleaching of individual fluorophore. This technology has now been used extensively for single molecule counting of protein, RNA, and other macromolecules in a variety of complexes or nanostructures. Here, we elucidate the SMPB technology, using the counting of RNA molecules within a bacteriophage phi29 DNA-packaging biomotor as an example. The method described here can be applied to the single molecule counting of other molecules in other systems. The construction of a concise, simple and economical single molecule total internal reflection fluorescence (TIRF) microscope combining prism-type and objective-type TIRF is described. The imaging system contains a deep-cooled sensitive EMCCD camera with single fluorophore detection sensitivity, a laser combiner for simultaneous dual-color excitation, and a Dual-View™ imager to split the multiple outcome signals to different detector channels based on their wavelengths. Methodology of the single molecule photobleaching assay used to elucidate the stoichiometry of RNA on phi29 DNA packaging motor and the mechanism of protein/RNA interaction are described. Different methods for single fluorophore labeling of RNA molecules are reviewed. The process of statistical modeling to reveal the true copy number of the biomolecules based on binomial distribution is also described. Copyright © 2014 Elsevier Inc. All rights reserved.
Reaction and Aggregation Dynamics of Cell Surface Receptors
NASA Astrophysics Data System (ADS)
Wang, Michelle Dong
This dissertation is composed of both theoretical and experimental studies of cell surface receptor reaction and aggregation. Project I studies the reaction rate enhancement due to surface diffusion of a bulk dissolved ligand with its membrane embedded target, using numerical calculations. The results show that the reaction rate enhancement is determined by ligand surface adsorption and desorption kinetic rates, surface and bulk diffusion coefficients, and geometry. In particular, we demonstrate that the ligand surface adsorption and desorption kinetic rates, rather than their ratio (the equilibrium constant), are important in rate enhancement. The second and third projects are studies of acetylcholine receptor clusters on cultured rat myotubes using fluorescence techniques after labeling the receptors with tetramethylrhodamine -alpha-bungarotoxin. The second project studies when and where the clusters form by making time-lapse movies. The movies are made from overlay of the pseudocolored total internal reflection fluorescence (TIRF) images of the cluster, and the schlieren images of the cell cultures. These movies are the first movies made using TIRF, and they clearly show the cluster formation from the myoblast fusion, the first appearance of clusters, and the eventual disappearance of clusters. The third project studies the fine structural features of individual clusters observed under TIRF. The features were characterized with six parameters by developing a novel fluorescence technique: spatial fluorescence autocorrelation. These parameters were then used to study the feature variations with age, and with treatments of drugs (oligomycin and carbachol). The results show little variation with age. However, drug treatment induced significant changes in some parameters. These changes were different for oligomycin and carbachol, which indicates that the two drugs may eliminate clusters through different mechanisms.
Nikolaus, Joerg; Karatekin, Erdem
2016-01-01
In the ubiquitous process of membrane fusion the opening of a fusion pore establishes the first connection between two formerly separate compartments. During neurotransmitter or hormone release via exocytosis, the fusion pore can transiently open and close repeatedly, regulating cargo release kinetics. Pore dynamics also determine the mode of vesicle recycling; irreversible resealing results in transient, "kiss-and-run" fusion, whereas dilation leads to full fusion. To better understand what factors govern pore dynamics, we developed an assay to monitor membrane fusion using polarized total internal reflection fluorescence (TIRF) microscopy with single molecule sensitivity and ~15 msec time resolution in a biochemically well-defined in vitro system. Fusion of fluorescently labeled small unilamellar vesicles containing v-SNARE proteins (v-SUVs) with a planar bilayer bearing t-SNAREs, supported on a soft polymer cushion (t-SBL, t-supported bilayer), is monitored. The assay uses microfluidic flow channels that ensure minimal sample consumption while supplying a constant density of SUVs. Exploiting the rapid signal enhancement upon transfer of lipid labels from the SUV to the SBL during fusion, kinetics of lipid dye transfer is monitored. The sensitivity of TIRF microscopy allows tracking single fluorescent lipid labels, from which lipid diffusivity and SUV size can be deduced for every fusion event. Lipid dye release times can be much longer than expected for unimpeded passage through permanently open pores. Using a model that assumes retardation of lipid release is due to pore flickering, a pore "openness", the fraction of time the pore remains open during fusion, can be estimated. A soluble marker can be encapsulated in the SUVs for simultaneous monitoring of lipid and soluble cargo release. Such measurements indicate some pores may reseal after losing a fraction of the soluble cargo. PMID:27585113
Optical sensing: recognition elements and devices
NASA Astrophysics Data System (ADS)
Gauglitz, Guenter G.
2012-09-01
The requirements in chemical and biochemical sensing with respect to recognition elements, avoiding non-specific interactions, and high loading of the surface for detection of low concentrations as well as optimized detection systems are discussed. Among the many detection principles the optical techniques are classified. Methods using labeled compounds like Total Internal Reflection Fluorescence (TIRF) and direct optical methods like micro reflectometry or refractometry are discussed in comparison. Reflectometric Interference Spectroscopy (RIfS) is presented as a robust simple method for biosensing. As applications, trace analysis of endocrine disruptors in water, hormones in food, detection of viruses and bacteria in food and clinical diagnostics are discussed.
Towards fully automated Identification of Vesicle-Membrane Fusion Events in TIRF Microscopy
NASA Astrophysics Data System (ADS)
Vallotton, Pascal; James, David E.; Hughes, William E.
2007-11-01
Total Internal Reflection Fluorescence Microscopy (TIRFM) is imposing itself as the tool of choice for studying biological activity in close proximity to the plasma membrane. For example, the exquisite selectivity of TIRFM allows monitoring the diffusion of GFP-phogrin vesicles and their recruitment to the plasma membrane in pancreatic β-cells. We present a novel computer vision system for automatically identifying the elusive fusion events of GFP-phogrin vesicles with the plasma membrane. Our method is based on robust object tracking and matched filtering. It should accelerate the quantification of TIRFM data and allow the extraction of more biological information from image data to support research in diabetes and obesity.
van 't Hoff, Marcel; Reuter, Marcel; Dryden, David T F; Oheim, Martin
2009-09-21
Bacteriophage lambda-DNA molecules are frequently used as a scaffold to characterize the action of single proteins unwinding, translocating, digesting or repairing DNA. However, scaling up such single-DNA-molecule experiments under identical conditions to attain statistically relevant sample sizes remains challenging. Additionally the movies obtained are frequently noisy and difficult to analyse with any precision. We address these two problems here using, firstly, a novel variable-angle total internal reflection fluorescence (VA-TIRF) reflector composed of a minimal set of optical reflective elements, and secondly, using single value decomposition (SVD) to improve the signal-to-noise ratio prior to analysing time-lapse image stacks. As an example, we visualize under identical optical conditions hundreds of surface-tethered single lambda-DNA molecules, stained with the intercalating dye YOYO-1 iodide, and stretched out in a microcapillary flow. Another novelty of our approach is that we arrange on a mechanically driven stage several capillaries containing saline, calibration buffer and lambda-DNA, respectively, thus extending the approach to high-content, high-throughput screening of single molecules. Our length measurements of individual DNA molecules from noise-reduced kymograph images using SVD display a 6-fold enhanced precision compared to raw-data analysis, reaching approximately 1 kbp resolution. Combining these two methods, our approach provides a straightforward yet powerful way of collecting statistically relevant amounts of data in a semi-automated manner. We believe that our conceptually simple technique should be of interest for a broader range of single-molecule studies, well beyond the specific example of lambda-DNA shown here.
Aoyagi, Kyota; Ohara-Imaizumi, Mica; Nishiwaki, Chiyono; Nakamichi, Yoko; Nagamatsu, Shinya
2009-01-01
To investigate the different effects between sulfonylurea (SU) and glinide drugs in insulin secretion, pancreatic β-cells were repeatedly stimulated with SU (glimepiride) or glinide (mitiglinide). Total internal reflection fluorescent (TIRF) microscopy revealed that secondary stimulation with glimepiride, but not glucose and mitiglinide, failed to evoke fusions of insulin granules although primary stimulation with glucose, glimepiride, and mitiglinide induced equivalent numbers of exocytotic responses. Glimepiride, but not glucose and mitiglinide, induced abnormally sustained [Ca2+]i elevations and reductions of docked insulin granules on the plasma membrane. Our data suggest that the effect of glinide on insulin secretory mechanisms is similar to that of glucose. PMID:20069052
Hemorrhagic Fever Virus Budding Studies.
Harty, Ronald N
2018-01-01
Independent expression of the VP40 or Z matrix proteins of filoviruses (marburgviruses and ebolaviruses) and arenaviruses (Lassa fever and Junín), respectively, gives rise to the production and release of virus-like particles (VLPs) that are morphologically identical to infectious virions. We can detect and quantify VLP production and egress in mammalian cells by transient transfection, SDS-PAGE, Western blotting, and live cell imaging techniques such as total internal reflection fluorescence (TIRF) microscopy. Since the VLP budding assay accurately mimics budding of infectious virus, this BSL-2 assay is safe and useful for the interrogation of both viral and host determinants required for budding and can be used as an initial screen to identify and validate small molecule inhibitors of virus release and spread.
A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Young, Laurence J.; Ströhl, Florian; Kaminski, Clemens F.
2016-01-01
Optical super-resolution imaging with structured illumination microscopy (SIM) is a key technology for the visualization of processes at the molecular level in the chemical and biomedical sciences. Although commercial SIM systems are available, systems that are custom designed in the laboratory can outperform commercial systems, the latter typically designed for ease of use and general purpose applications, both in terms of imaging fidelity and speed. This article presents an in-depth guide to building a SIM system that uses total internal reflection (TIR) illumination and is capable of imaging at up to 10 Hz in three colors at a resolution reaching 100 nm. Due to the combination of SIM and TIRF, the system provides better image contrast than rival technologies. To achieve these specifications, several optical elements are used to enable automated control over the polarization state and spatial structure of the illumination light for all available excitation wavelengths. Full details on hardware implementation and control are given to achieve synchronization between excitation light pattern generation, wavelength, polarization state, and camera control with an emphasis on achieving maximum acquisition frame rate. A step-by-step protocol for system alignment and calibration is presented and the achievable resolution improvement is validated on ideal test samples. The capability for video-rate super-resolution imaging is demonstrated with living cells. PMID:27285848
Single-Molecule Three-Color FRET with Both Negligible Spectral Overlap and Long Observation Time
Hohng, Sungchul
2010-01-01
Full understanding of complex biological interactions frequently requires multi-color detection capability in doing single-molecule fluorescence resonance energy transfer (FRET) experiments. Existing single-molecule three-color FRET techniques, however, suffer from severe photobleaching of Alexa 488, or its alternative dyes, and have been limitedly used for kinetics studies. In this work, we developed a single-molecule three-color FRET technique based on the Cy3-Cy5-Cy7 dye trio, thus providing enhanced observation time and improved data quality. Because the absorption spectra of three fluorophores are well separated, real-time monitoring of three FRET efficiencies was possible by incorporating the alternating laser excitation (ALEX) technique both in confocal microscopy and in total-internal-reflection fluorescence (TIRF) microscopy. PMID:20808851
NASA Astrophysics Data System (ADS)
Barroso Peña, Álvaro; Nieves, Marcos; Teper, Konrad; Wedlich-Soldner, Roland; Denz, Cornelia
2016-09-01
The plasma membrane serves as protective interface between cells and their environment. It also constitutes a hub for selective nutrient uptake and signal transduction. Increasing evidence over the last years indicates that, similar to eukaryotic cells, lateral membrane organization plays an important role in the regulation of prokaryotic signaling pathways. However, the mechanisms underlying this phenomenon are still poorly understood. Spatiotemporal characterization of bacterial signal transduction demands very sensitive high-resolution microscopy techniques due to the low expression levels of most signaling proteins and the small size of bacterial cells. In addition, direct study of subcellular confinement and dynamics of bacterial signaling proteins during the different stages of the signal transduction also requires immobilization in order to avoid cell displacement caused by Brownian motion, local fluid flows and bacterial self-propulsion. In this work we present a novel approach based on the combination of high resolution imaging and optical manipulation that enables the investigation of the distribution and dynamics of proteins at the bacterial plasma membrane. For this purpose, we combine the versatility of holographic optical tweezers (HOT) with the sensitivity and resolution of total internal reflection fluorescence (TIRF) microscopy. Furthermore, we discuss the implementation of microfluidic devices in our integrated HOT+TIRF system for the control of growth conditions of bacterial cells. The capabilities of our workstation provides thus new valuable insights into the fundamental cellular and physical mechanisms underlying the regulation of bacterial signal transduction.
Ding, Yong-Xue; Streitmatter, Seth; Wright, Bryon E.; Hlady, Vladimir
2010-01-01
A gradient of negative surface charge based on 1-D spatial variation from surface sulfhydryl to mixed sulfhydryl-sulfonate moities was prepared by controlled UV oxidation of 3-mercaptopropylsilane monolayer on fused silica. Adsorption of three human plasma proteins, albumin (HSA), immunoglobulin G (IgG), and fibrinogen (Fgn) onto such surface gradient was studied using spatially-resolved total internal reflection fluorescence (TIRF) and autoradiography. Adsorption was measured from dilute solutions equivalent to 1/100 (TIRF, autoradiography), and 1/500 and 1/1000 (autoradiography) of protein’s physiological concentrations in plasma. All three proteins adsorbed more to the non-oxidized sulfhydryl region than to the oxidized, mixed sulfhydryl-sulfonate region of the gradient. In the case of HSA the adsorption contrast along the gradient was largest when the adsorption took place from more dilute protein solutions. Increasing the concentration to 1/100 of protein plasma concentration eliminated the effect of the gradient on HSA adsorption and to the lesser extent on IgG adsorption. In the case of Fgn the greatest adsorption contrast was observed at the highest concentration used. Based on adsorption kinetics, the estimated binding affinity of HSA for the sulfhydryl region what twice the affinity for the mixed sulfhydryl-sulfonate region of the gradient. For IgG and Fgn the initial adsorption was transport-limited and the initial adsorption rates approached the computed flux of the protein to the surface. PMID:20568822
Trache, Andreea; Meininger, Gerald A
2005-01-01
A novel hybrid imaging system is constructed integrating atomic force microscopy (AFM) with a combination of optical imaging techniques that offer high spatial resolution. The main application of this instrument (the NanoFluor microscope) is the study of mechanotransduction with an emphasis on extracellular matrix-integrin-cytoskeletal interactions and their role in the cellular responses to changes in external chemical and mechanical factors. The AFM allows the quantitative assessment of cytoskeletal changes, binding probability, adhesion forces, and micromechanical properties of the cells, while the optical imaging applications allow thin sectioning of the cell body at the coverslip-cell interface, permitting the study of focal adhesions using total internal reflection fluorescence (TIRF) and internal reflection microscopy (IRM). Combined AFM-optical imaging experiments show that mechanical stimulation at the apical surface of cells induces a force-generating cytoskeletal response, resulting in focal contact reorganization on the basal surface that can be monitored in real time. The NanoFluor system is also equipped with a novel mechanically aligned dual camera acquisition system for synthesized Forster resonance energy transfer (FRET). The integrated NanoFluor microscope system is described, including its characteristics, applications, and limitations.
Evanescent field microscopy techniques for studying dynamics at the surface of living cells
NASA Astrophysics Data System (ADS)
Sund, Susan E.
This thesis presents two distinct optical microscopy techniques for applications in cell biophysics: (a)the extension to living cells of an established technique, total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP) for the first time in imaging mode; and (b)the novel development of polarized total internal reflection fluorescence (p- TIRF) to study membrane orientation in living cells. Although reversible chemistry is crucial to dynamical processes in living cells, relatively little is known about the relevant chemical kinetic rates in vivo. TIR/FRAP, an established technique which can measure reversible biomolecular kinetic rates at surfaces, is extended here to measure kinetic parameters of microinjected rhodamine actin at the cytofacial surface of the plasma membrane of living cultured smooth muscle cells. For the first time, spatial imaging (with a CCD camera) is used in conjunction with TIR/FRAP. TIR/FRAP imaging allows production of spatially resolved images of kinetic data, and calculation of correlation distances, cell-wide gradients, and kinetic parameter dependence on initial fluorescence intensity. In living cells, membrane curvature occurs both in easily imaged large scale morphological features, and also in less visualizable submicroscopic regions of activity such as endocytosis, exocytosis, and cell surface ruffling. A fluorescence microscopic method, p-TIRF, is introduced here to visualize such regions. The method is based on fluorescence of the oriented membrane probe diI- C18-(3) (diI) excited by evanescent field light polarized either perpendicular or parallel to the plane of the substrate coverslip. The excitation efficiency from each polarization depends on the membrane orientation, and thus the ratio of the observed fluorescence excited by these two polarizations vividly shows regions of microscopic and submicroscopic curvature of the membrane. A theoretical background of the technique and experimental verifications are presented in samples of protein solutions, model lipid bilayers, and living cells. Sequential digital images of the polarized TIR fluorescence ratios show spatially-resolved time- course maps of membrane orientations on diI labeled macrophages from which low visibility membrane structures can be identified and quantified. The TIR images are sharpened and contrast-enhanced by deconvoluting them with an experimentally-measured point spread function.
Mishra, Anurag; Mishra, Ritu; Gottschalk, Sven; Pal, Robert; Sim, Neil; Engelmann, Joern; Goldberg, Martin; Parker, David
2014-02-19
A series of bimodal metabotropic glutamate-receptor targeted MRI contrast agents has been developed and evaluated, based on established competitive metabotropic Glu receptor subtype 5 (mGluR5) antagonists. In order to directly visualize mGluR5 binding of these agents on the surface of live astrocytes, variations in the core structure were made. A set of gadolinium conjugates containing either a cyanine dye or a fluorescein moiety was accordingly prepared, to allow visualization by optical microscopy in cellulo. In each case, surface receptor binding was compromised and cell internalization observed. Another approach, examining the location of a terbium analogue via sensitized emission, also exhibited nonspecific cell uptake in neuronal cell line models. Finally, biotin derivatives of two lead compounds were prepared, and the specificity of binding to the mGluR5 cell surface receptors was demonstrated with the aid of their fluorescently labeled avidin conjugates, using both total internal reflection fluorescence (TIRF) and confocal microscopy.
Single-Molecule Analysis for RISC Assembly and Target Cleavage.
Sasaki, Hiroshi M; Tadakuma, Hisashi; Tomari, Yukihide
2018-01-01
RNA-induced silencing complex (RISC) is a small RNA-protein complex that mediates silencing of complementary target RNAs. Biochemistry has been successfully used to characterize the molecular mechanism of RISC assembly and function for nearly two decades. However, further dissection of intermediate states during the reactions has been warranted to fill in the gaps in our understanding of RNA silencing mechanisms. Single-molecule analysis with total internal reflection fluorescence (TIRF) microscopy is a powerful imaging-based approach to interrogate complex formation and dynamics at the individual molecule level with high sensitivity. Combining this technique with our recently established in vitro reconstitution system of fly Ago2-RISC, we have developed a single-molecule observation system for RISC assembly. In this chapter, we summarize the detailed protocol for single-molecule analysis of chaperone-assisted assembly of fly Ago2-RISC as well as its target cleavage reaction.
Sub-micrometer Geometrically Encoded Fluorescent Barcodes Self-Assembled from DNA
Lin, Chenxiang; Jungmann, Ralf; Leifer, Andrew M.; Li, Chao; Levner, Daniel; Church, George M.; Shih, William M.; Yin, Peng
2012-01-01
The identification and differentiation of a large number of distinct molecular species with high temporal and spatial resolution is a major challenge in biomedical science. Fluorescence microscopy is a powerful tool, but its multiplexing ability is limited by the number of spectrally distinguishable fluorophores. Here we use DNA-origami technology to construct sub-micrometer nanorods that act as fluorescent barcodes. We demonstrate that spatial control over the positioning of fluorophores on the surface of a stiff DNA nanorod can produce 216 distinct barcodes that can be unambiguously decoded using epifluorescence or total internal reflection fluorescence (TIRF) microscopy. Barcodes with higher spatial information density were demonstrated via the construction of super-resolution barcodes with features spaced by ~40 nm. One species of the barcodes was used to tag yeast surface receptors, suggesting their potential applications as in situ imaging probes for diverse biomolecular and cellular entities in their native environments. PMID:23000997
Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist.
Sanchez, Elisa; Huse, Morgan
2018-04-25
T lymphocytes engage in rapid, polarized signaling, occurring within minutes following TCR activation. This induces formation of the immunological synapse, a stereotyped cell-cell junction that regulates T cell activation and directionally targets effector responses. To study these processes effectively, an imaging approach that is tailored to capturing fast, polarized responses is necessary. This protocol describes such a system, which is based on a photoactivatable peptide-major histocompatibility complex (pMHC) that is non-stimulatory until it is exposed to ultraviolet light. Targeted decaging of this reagent during videomicroscopy experiments enables precise spatiotemporal control of TCR activation and high-resolution monitoring of subsequent cellular responses by total internal reflection (TIRF) imaging. This approach is also compatible with genetic and pharmacological perturbation strategies. This allows for the assembly of well-defined molecular pathways that link TCR signaling to the formation of the polarized cytoskeletal structures that underlie the immunological synapse.
Kinetic Characterization of Nonmuscle Myosin IIB at the Single Molecule Level*
Nagy, Attila; Takagi, Yasuharu; Billington, Neil; Sun, Sara A.; Hong, Davin K. T.; Homsher, Earl; Wang, Aibing; Sellers, James R.
2013-01-01
Nonmuscle myosin IIB (NMIIB) is a cytoplasmic myosin, which plays an important role in cell motility by maintaining cortical tension. It forms bipolar thick filaments with ∼14 myosin molecule dimers on each side of the bare zone. Our previous studies showed that the NMIIB is a moderately high duty ratio (∼20–25%) motor. The ADP release step (∼0.35 s−1) of NMIIB is only ∼3 times faster than the rate-limiting phosphate release (0.13 ± 0.01 s−1). The aim of this study was to relate the known in vitro kinetic parameters to the results of single molecule experiments and to compare the kinetic and mechanical properties of single- and double-headed myosin fragments and nonmuscle IIB thick filaments. Examination of the kinetics of NMIIB interaction with actin at the single molecule level was accomplished using total internal reflection fluorescence (TIRF) with fluorescence imaging with 1-nm accuracy (FIONA) and dual-beam optical trapping. At a physiological ATP concentration (1 mm), the rate of detachment of the single-headed and double-headed molecules was similar (∼0.4 s−1). Using optical tweezers we found that the power stroke sizes of single- and double-headed heavy meromyosin (HMM) were each ∼6 nm. No signs of processive stepping at the single molecule level were observed in the case of NMIIB-HMM in optical tweezers or TIRF/in vitro motility experiments. In contrast, robust motility of individual fluorescently labeled thick filaments of full-length NMIIB was observed on actin filaments. Our results are in good agreement with the previous steady-state and transient kinetic studies and show that the individual nonprocessive nonmuscle myosin IIB molecules form a highly processive unit when polymerized into filaments. PMID:23148220
Chahinian, Henri; Snabe, Torben; Attias, Coralie; Fojan, Peter; Petersen, Steffen B; Carrière, Frédéric
2006-01-24
Gastric lipase is active under acidic conditions and shows optimum activity on insoluble triglycerides at pH 4. The present results show that gastric lipase also acts in solution on vinyl butyrate, with an optimum activity above pH 7, which suggests that gastric lipase is able to hydrolyze ester bonds via the classical mechanism of serine hydrolases. These results support previous structural studies in which the catalytic triad of gastric lipase was reported to show no specific features. The optimum activity of gastric lipase shifted toward lower pH values, however, when the vinyl butyrate concentration was greater than the solubility limit. Experiments performed with long-chain triglycerides showed that gastric lipase binds optimally to the oil-water interface at low pH values. To study the effects of the pH on the adsorption step independently from substrate hydrolysis, gastric lipase adsorption on solid hydrophobic surfaces was monitored by total internal reflection fluorescence (TIRF), as well as using a quartz crystal microbalance. Both techniques showed a pH-dependent reversible gastric lipase adsorption process, which was optimum at pH 5 (Kd = 6.5 nM). Lipase adsorption and desorption constants (ka = 147,860 M(-1) s(-1) and kd = 139 x 10(-4) s(-1) at pH 6) were estimated from TIRF experiments. These results indicate that the optimum activity of gastric lipase at acidic pH is only "apparent" and results from the fact that lipase adsorption at lipid-water interfaces is the pH-dependent limiting step in the overall process of insoluble substrate hydrolysis. This specific kinetic feature of interfacial enzymology should be taken into account when studying any soluble enzyme acting on an insoluble substrate.
Coalescence driven self-organization of growing nanodroplets around a microcap
NASA Astrophysics Data System (ADS)
Dyett, Brendan; Hao, Hao; Lohse, Detlef; Zhang, Xuehua
The coalescence between growing droplets is important for the surface coverage and spatial arrangements of droplets on surfaces. In this work, total internal reflection fluorescence (TIRF) microscopy is utilized to in-situ investigate the formation of nanodroplets around the rim of a polymer microcap, with sub-micron spatial and millisecond temporal resolution. We observe that the coalescence among droplets occurs frequently during their growth by solvent exchange. Our experimental results show that the position of the droplet from two merged droplets is related to the size of the parent droplets. The position of the coalesced droplet and the ratio of parent droplet sizes obey a scaling law, reflecting a coalescence preference based on the size inequality. As a result of droplet coalescence, the angles between the centroids of two neighbouring droplets increase with time, obeying a nearly symmetrical arrangement of droplets at various time intervals. The evolution of the position and number from coalescence of growing droplets is modelled. The mechanism for coalescence driven self-organization of growing droplets is general, applicable to microcaps of different sizes and droplets of different liquids. The understanding from this work may be valuable for positioning nanodroplets by nucleation and growth without using templates.
Coalescence driven self-organization of growing nanodroplets around a microcap.
Dyett, Brendan; Hao, Hao; Lohse, Detlef; Zhang, Xuehua
2018-04-04
The coalescence between growing droplets is important for the surface coverage and spatial arrangements of droplets on surfaces. In this work, total internal reflection fluorescence (TIRF) microscopy is utilized to in situ investigate the formation of nanodroplets around the rim of a polymer microcap, with sub-micron spatial and millisecond temporal resolution. We observe that the coalescence among droplets occurs frequently during their growth by solvent exchange. Our experimental results show that the position of the droplet from two merged droplets is related to the size of the parent droplets. The position of the coalesced droplet and the ratio of parent droplet sizes obey a scaling law, reflecting a coalescence preference based on the size inequality. As a result of droplet coalescence, the angles between the centroids of two neighbouring droplets increase with time, obeying a nearly symmetrical arrangement of droplets at various time intervals. The evolution of the position and number from coalescence of growing droplets is modelled. The mechanism for coalescence driven self-organization of growing droplets is general, applicable to microcaps of different sizes and droplets of different liquids. The understanding from this work may be valuable for positioning nanodroplets by nucleation and growth without using templates.
Using Amphiphilic Copolymers and Nanoparticles to Organize Charged Biopolymers
NASA Astrophysics Data System (ADS)
Park, Jung Hyun; McConnell, Marla; Sun, Yujie; Goldman, Yale; Composto, Russell
2009-03-01
Nanoparticles (NPs) on amphiphilic random copolymers control filamentous actin (F-actin) attachment. 3-aminopropyltriethoxysilane (APTES) coated silica NPs are selectively bonded to acrylic acid groups on the surface of a poly(styrene-r-acrylic acid) (PS-r-PAA) film. By changing the concentration of NPs in the medium, the surface density of positively charged anchors is tuned. Using total internal reflection fluorescence (TIRF) microscopy, immobilization of F-actin is observed via electrostatic interaction with NPs at high NP coverages. Below a critical coverage, F-actin is weakly attached and undergoes thermal fluctuations near the surface. Another method to tune F-actin attachment is to use APTES to cross-link and create positive charge in PAA films. Here, the surface coverage of F-actin decreases as APTES concentration increases. This observation is attributed to an increase in surface roughness and hydrophobicity that reduces the effective surface sites that attract F-actin. In addition, in-situ G-actin polymerization to F-actin is observed on both the NP and cross-linked PAA templates.
Zhang, Liangyu; Shao, Hengyi; Zhu, Tongge; Xia, Peng; Wang, Zhikai; Liu, Lifang; Yan, Maomao; Hill, Donald L.; Fang, Guowei; Chen, Zhengjun; Wang, Dongmei; Yao, Xuebiao
2013-01-01
Cell motility and adhesion involve orchestrated interaction of microtubules (MTs) with their plus-end tracking proteins (+TIPs). However, the mechanisms underlying regulations of MT dynamics and directional cell migration are still elusive. Here, we show that DDA3-EB1 interaction orchestrates MT plus-end dynamics and facilitates directional cell migration. Biochemical characterizations reveal that DDA3 interacts with EB1 via its SxIP motif within the C-terminal Pro/Ser-rich region. Time-lapse and total internal reflection fluorescence (TIRF) microscopic assays demonstrate that DDA3 exhibits EB1-dependent, MT plus-end loading and tracking. The EB1-based loading of DDA3 is responsible for MT plus-ends stabilization at the cell cortex, which in turn orchestrates directional cell migration. Interestingly, the DDA3-EB1 interaction is potentially regulated by EB1 acetylation, which may account for physiological regulation underlying EGF-elicited cell migration. Thus, the EB1-based function of DDA3 links MT dynamics to directional cell migration. PMID:23652583
Mutch, Sarah A.; Gadd, Jennifer C.; Fujimoto, Bryant S.; Kensel-Hammes, Patricia; Schiro, Perry G.; Bajjalieh, Sandra M.; Chiu, Daniel T.
2013-01-01
This protocol describes a method to determine both the average number and variance of proteins in the few to tens of copies in isolated cellular compartments, such as organelles and protein complexes. Other currently available protein quantification techniques either provide an average number but lack information on the variance or are not suitable for reliably counting proteins present in the few to tens of copies. This protocol entails labeling the cellular compartment with fluorescent primary-secondary antibody complexes, TIRF (total internal reflection fluorescence) microscopy imaging of the cellular compartment, digital image analysis, and deconvolution of the fluorescence intensity data. A minimum of 2.5 days is required to complete the labeling, imaging, and analysis of a set of samples. As an illustrative example, we describe in detail the procedure used to determine the copy number of proteins in synaptic vesicles. The same procedure can be applied to other organelles or signaling complexes. PMID:22094731
Bonham, Andrew J.; Wenta, Nikola; Osslund, Leah M.; Prussin, Aaron J.; Vinkemeier, Uwe; Reich, Norbert O.
2013-01-01
The DNA-binding specificity and affinity of the dimeric human transcription factor (TF) STAT1, were assessed by total internal reflectance fluorescence protein-binding microarrays (TIRF-PBM) to evaluate the effects of protein phosphorylation, higher-order polymerization and small-molecule inhibition. Active, phosphorylated STAT1 showed binding preferences consistent with prior characterization, whereas unphosphorylated STAT1 showed a weak-binding preference for one-half of the GAS consensus site, consistent with recent models of STAT1 structure and function in response to phosphorylation. This altered-binding preference was further tested by use of the inhibitor LLL3, which we show to disrupt STAT1 binding in a sequence-dependent fashion. To determine if this sequence-dependence is specific to STAT1 and not a general feature of human TF biology, the TF Myc/Max was analysed and tested with the inhibitor Mycro3. Myc/Max inhibition by Mycro3 is sequence independent, suggesting that the sequence-dependent inhibition of STAT1 may be specific to this system and a useful target for future inhibitor design. PMID:23180800
Light, Imaging, Vision: An interdisciplinary undergraduate course
NASA Astrophysics Data System (ADS)
Nelson, Philip
2015-03-01
The vertebrate eye is fantastically sensitive instrument, capable of registering the absorption of a single photon, and yet generating very low noise. Using eyes as a common thread helps motivate undergraduates to learn a lot of physics, both fundamental and applied to scientific imaging and neuroscience. I'll describe an undergraduate course, for students in several science and engineering majors, that takes students from the rudiments of probability theory to the quantum character of light, including modern experimental methods like fluorescence imaging and Förster resonance energy transfer. After a digression into color vision, we then see how the Feynman principle explains the apparently wavelike phenomena associated to light, including applications like diffraction, subdiffraction imaging, total internal reflection and TIRF microscopy. Then we see how scientists documented the single-quantum sensitivity of the eye seven decades earlier than ``ought'' to have been possible, and finally close with the remarkable signaling cascade that delivers such outstanding performance. Parts of this story are now embodied in a new textbook (WH Freeman and Co, 1/2015); additional course materials are available upon request. Work supported by NSF Grants EF-0928048 and DMR-0832802.
Imaging intracellular protein dynamics by spinning disk confocal microscopy
Stehbens, Samantha; Pemble, Hayley; Murrow, Lindsay; Wittmann, Torsten
2012-01-01
The palette of fluorescent proteins has grown exponentially over the last decade, and as a result live imaging of cells expressing fluorescently tagged proteins is becoming more and more main stream. Spinning disk confocal microscopy (SDC) is a high speed optical sectioning technique, and a method of choice to observe and analyze intracellular fluorescent protein dynamics at high spatial and temporal resolution. In an SDC system, a rapidly rotating pinhole disk generates thousands of points of light that scan the specimen simultaneously, which allows direct capture of the confocal image with low noise scientific grade cooled charged-coupled device (CCD) cameras, and can achieve frame rates of up 1000 frames per second. In this chapter we describe important components of a state-of-the-art spinning disk system optimized for live cell microscopy, and provide a rationale for specific design choices. We also give guidelines how other imaging techniques such as total internal reflection (TIRF) microscopy or spatially controlled photoactivation can be coupled with SDC imaging, and provide a short protocol on how to generate cell lines stably expressing fluorescently tagged proteins by lentivirus-mediated transduction. PMID:22264541
Quantifying the Dynamic Interactions Between a Clathrin-Coated Pit and Cargo Molecules
NASA Astrophysics Data System (ADS)
Weigel, Aubrey; Tamkun, Michael; Krapf, Diego
2014-03-01
Clathrin-mediated endocytosis is a major pathway of internalization of cargo in eukaryotic cells. This process involves the recruitment of cargo molecules into a growing clathrin-coated pit (CCP). However, cargo-CCP interactions are difficult to study because CCPs display a large degree of lifetime heterogeneity and the interactions with cargo molecules evolve over time. We use single-molecule total internal reflection fluorescence (TIRF) microscopy, in combination with automatic detection and tracking algorithms, to directly visualize the recruitment of individual voltage-gated potassium channels into forming CCPs in living cells. Contrary to widespread ideas, cargo often escapes from a pit before abortive CCP termination or endocytic vesicle production. By measuring tens of thousands of capturing events, we build the distribution of capture times and the times that cargo remains confined to a CCP. An analytical stochastic model is developed and compared to the measured distributions. Due to the dynamic nature of the pit, the model is non-Markovian and it displays long-tail power law statistics. Our findings identify one source of the large heterogeneities observed in CCP maturation and provide a mechanism for the anomalous diffusion of proteins in the plasma membrane. This work was supported by National Science Foundation Grant PHY-0956714.
Zarcone, Jennifer; Hagopian, Louis; Ninci, Jennifer; McKay, Chloe; Bonner, Andrew; Dillon, Christopher; Hausman, Nicole
2016-01-01
The goal of this study was to develop and evaluate a tool to measure the complexity and intensity of psychotropic medication interventions, behavioral interventions, and issues related to crisis management for challenging behavior using a standardized rating form. The Treatment Intensity Rating Form (TIRF) is a 10-item scale with three categories: pharmacological interventions, behavior supports, and protective equipment. In a retrospective review we examined the final treatment recommendations for 74 individuals with self-injurious behavior (SIB) based on psychiatric and behavioral notes and reports. We also compared whether TIRF scores differed across individuals for whom SIB was maintained by social reinforcement (e.g., to access attention or toys/activities, or escape from tasks) versus those for whom SIB was maintained by automatic reinforcement (e.g., occurs independent of social variables, and is presumed to be maintained by sensory reinforcement). The TIRF was demonstrated to have strong inter-rater reliability (98%) and appears to have good face validity. As hypothesized, individuals with SIB maintained by automatic reinforcement had significantly more medication trials (p=0.0005) and required more protective equipment than individuals with SIB maintained by social reinforcement (p=0.0002). Antidepressant medication was used more often with individuals with automatically reinforced SIB, although antipsychotics and anticonvulsants were also commonly used across both groups. Findings provide initial support for the TIRF's reliability, and face validity as a measure the level of complexity of medical and behavioral treatment plans - although additional research is needed to fully evaluate its psychometric properties.
Czeslik, C.; Royer, C.; Hazlett, T.; Mantulin, W.
2003-01-01
The preservation of enzyme activity and protein binding capacity upon protein adsorption at solid interfaces is important for biotechnological and medical applications. Because these properties are partly related to the protein flexibility and mobility, we have studied the internal dynamics and the whole-body reorientational rates of two enzymes, staphylococcal nuclease (SNase) and hen egg white lysozyme, over the temperature range of 20–80°C when the proteins are adsorbed at the silica/water interface and, for comparison, when they are dissolved in buffer. The data were obtained using a combination of two experimental techniques, total internal reflection fluorescence spectroscopy and time-resolved fluorescence anisotropy measurements in the frequency domain, with the protein Trp residues as intrinsic fluorescence probes. It has been found that the internal dynamics and the whole-body rotation of SNase and lysozyme are markedly reduced upon adsorption over large temperature ranges. At elevated temperatures, both protein molecules appear completely immobilized and the fractional amplitudes for the whole-body rotation, which are related to the order parameter for the local rotational freedom of the Trp residues, remain constant and do not approach zero. This behavior indicates that the angular range of the Trp reorientation within the adsorbed proteins is largely restricted even at high temperatures, in contrast to that of the dissolved proteins. The results of this study thus provide a deeper understanding of protein activity at solid surfaces. PMID:12668461
Redistribution of caveolae during mitosis
Boucrot, Emmanuel; Howes, Mark T.; Kirchhausen, Tomas; Parton, Robert G.
2011-01-01
Caveolae form a specialized platform within the plasma membrane that is crucial for an array of important biological functions, ranging from signaling to endocytosis. Using total internal reflection fluorescence (TIRF) and 3D fast spinning-disk confocal imaging to follow caveola dynamics for extended periods, and electron microscopy to obtain high resolution snapshots, we found that the vast majority of caveolae are dynamic with lifetimes ranging from a few seconds to several minutes. Use of these methods revealed a change in the dynamics and localization of caveolae during mitosis. During interphase, the equilibrium between the arrival and departure of caveolae from the cell surface maintains the steady-state distribution of caveolin-1 (Cav1) at the plasma membrane. During mitosis, increased dynamics coupled to an imbalance between the arrival and departure of caveolae from the cell surface induces a redistribution of Cav1 from the plasma membrane to intracellular compartments. These changes are reversed during cytokinesis. The observed redistribution of Cav1 was reproduced by treatment of interphase cells with nocodazole, suggesting that microtubule rearrangements during mitosis can mediate caveolin relocalization. This study provides new insights into the dynamics of caveolae and highlights precise regulation of caveola budding and recycling during mitosis. PMID:21625007
Light, Imaging, Vision: An interdisciplinary undergraduate course
NASA Astrophysics Data System (ADS)
Nelson, Philip
Students in physical and life science, and in engineering, need to know about the physics and biology of light. In the 21st century, it has become increasingly clear that the quantum nature of light is essential both for the latest imaging modalities and even to advance our knowledge of fundamental processes, such as photosynthesis and human vision. But many optics courses remain rooted in classical physics, with photons as an afterthought. I'll describe a new undergraduate course, for students in several science and engineering majors, that takes students from the rudiments of probability theory to modern methods like fluorescence imaging and Förster resonance energy transfer. After a digression into color vision, students then see how the Feynman principle explains the apparently wavelike phenomena associated to light, including applications like diffraction limit, subdiffraction imaging, total internal reflection and TIRF microscopy. Then we see how scientists documented the single-quantum sensitivity of the eye seven decades earlier than `ought' to have been possible, and finally close with the remarkable signaling cascade that delivers such outstanding performance. A new textbook embodying this course will be published by Princeton University Press in Spring 2017. Partially supported by the United States National Science Foundation under Grant PHY-1601894.
Kellogg, K.S.; Blank, H.R.
1982-01-01
Paleomagnetic directions determined for an upper Oligocene to lower Miocene dike swarm and from two lower Miocene layered gabbros in the Tihamat Asir coastal plain of southwestern Saudi Arabia are used to test several hypotheses concerning the tectonics of rifting along the eastern margin of the Red Sea. The dikes and gabbros were emplaced during the initial phases of Red Sea rifting and may mark the transition between continental and oceanic crust. Although these rocks have been hydrothermally altered to varying degrees, reliable remanent directions after alternating-field demagnetization were obtained for 23 dikes and for gabbros at Jabal at Tirf and Wadi Liyyah. Twelve of the dikes are reversely magnetized. After the directions of the reversely magnetized dikes are inverted 180?, the mean direction calculated for the normal dikes is approximately 24? more downward than that calculated for the reversed dikes. This result is similar to that found for the As Sarat volcanic field, 100 km to the north, and may be due to a displaced dipole source for the field. The unrotated mean remanent direction for the dikes (inverting reversed dike directions 180?) is D (declination) = 353.2? and I (inclination) = 6.8? with a95 (radius of the cone of 95 percent confidence) = 8.9? whereas directions from the Jabal at Tirf and Wadi Liyyah gabbros lie at D = 176.2?, I = -1.6? (a95 = 7 1 ?) and D = 17.1?, I = 16.3? (a 95 = 8.7?), respectively. Comparing these results with the results from the As Sarat volcanic field, all the paleomagnetic evidence supports a model for approximately 20 ? of westward tilting of the Wadi Damad and Wadi Jizan areas after the emplacement of the Jabal at Tiff gabbro. The Wadi Liyyah area may have been tilted even more toward the Red Sea. The paleomagnetic directions from three widely separated localities in the Jabal at Tirf gabbro are not significantly different, a fact which indicates that the body cooled in approximately its present bowl shape. Evidence suggests that the ratio of normal to reversed dikes may change significantly along a 6-km-long traverse normal to the trend of the dike swarm, possibly reflecting migration of .a spreading axis.
Single-particle fusion of influenza viruses reveals complex interactions with target membranes
NASA Astrophysics Data System (ADS)
van der Borg, Guus; Braddock, Scarlett; Blijleven, Jelle S.; van Oijen, Antoine M.; Roos, Wouter H.
2018-05-01
The first step in infection of influenza A virus is contact with the host cell membrane, with which it later fuses. The composition of the target bilayer exerts a complex influence on both fusion efficiency and time. Here, an in vitro, single-particle approach is used to study this effect. Using total internal reflection fluorescence (TIRF) microscopy and a microfluidic flow cell, the hemifusion of single virions is visualized. Hemifusion efficiency and kinetics are studied while altering target bilayer cholesterol content and sialic-acid donor. Cholesterol ratios tested were 0%, 10%, 20%, and 40%. Sialic-acid donors GD1a and GYPA were used. Both cholesterol ratio and sialic-acid donors proved to have a significant effect on hemifusion efficiency. Furthermore, comparison between GD1a and GYPA conditions shows that the cholesterol dependence of the hemifusion time is severely affected by the sialic-acid donor. Only GD1a shows a clear increasing trend in hemifusion efficiency and time with increasing cholesterol concentration of the target bilayer with maximum rates for GD1A and 40% cholesterol. Overall our results show that sialic acid donor and target bilayer composition should be carefully chosen, depending on the desired hemifusion time and efficiency in the experiment.
A single molecule study of cellulase hydrolysis of crystalline cellulose
NASA Astrophysics Data System (ADS)
Liu, Yu-San; Luo, Yonghua; Baker, John O.; Zeng, Yining; Himmel, Michael E.; Smith, Steve; Ding, Shi-You
2010-02-01
Cellobiohydrolase-I (CBH I), a processive exoglucanase secreted by Trichoderma reesei, is one of the key enzyme components in a commercial cellulase mixture currently used for processing biomass to biofuels. CBH I contains a family 7 glycoside hydrolase catalytic module, a family 1 carbohydrate-binding module (CBM), and a highlyglycosylated linker peptide. It has been proposed that the CBH I cellulase initiates the hydrolysis from the reducing end of one cellulose chain and successively cleaves alternate β-1,4-glycosidic bonds to release cellobiose as its principal end product. The role each module of CBH I plays in the processive hydrolysis of crystalline cellulose has yet to be convincingly elucidated. In this report, we use a single-molecule approach that combines optical (Total Internal Reflection Fluorescence microscopy, or TIRF-M) and non-optical (Atomic Force Microscopy, or AFM) imaging techniques to analyze the molecular motion of CBM tagged with green fluorescence protein (GFP), and to investigate the surface structure of crystalline cellulose and changes made in the structure by CBM and CBH I. The preliminary results have revealed a confined nanometer-scale movement of the TrCBM1-GFP bound to cellulose, and decreases in cellulose crystal size as well as increases in surface roughness during CBH I hydrolysis of crystalline cellulose.
Exogenous lysophospholipids with large head groups perturb clathrin-mediated endocytosis.
Ailte, Ieva; Lingelem, Anne Berit D; Kvalvaag, Audun S; Kavaliauskiene, Simona; Brech, Andreas; Koster, Gerbrand; Dommersnes, Paul G; Bergan, Jonas; Skotland, Tore; Sandvig, Kirsten
2017-03-01
In this study, we have investigated how clathrin-dependent endocytosis is affected by exogenously added lysophospholipids (LPLs). Addition of LPLs with large head groups strongly inhibits transferrin (Tf) endocytosis in various cell lines, while LPLs with small head groups do not. Electron and total internal reflection fluorescence microscopy (EM and TIRF) reveal that treatment with lysophosphatidylinositol (LPI) with the fatty acyl group C18:0 leads to reduced numbers of invaginated clathrin-coated pits (CCPs) at the plasma membrane, fewer endocytic events per membrane area and increased lifetime of CCPs. Also, endocytosis of Tf becomes dependent on actin upon LPI treatment. Thus, our results demonstrate that one can regulate the kinetics and properties of clathrin-dependent endocytosis by addition of LPLs in a head group size- and fatty acyl-dependent manner. Furthermore, studies performed with optical tweezers show that less force is required to pull membrane tubules outwards from the plasma membrane when LPI is added to the cells. The results are in agreement with the notion that insertion of LPLs with large head groups creates a positive membrane curvature which might have a negative impact on events that require plasma membrane invagination, while it may facilitate membrane bending toward the cell exterior. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Yamashiro, Sawako; Watanabe, Naoki
2017-01-01
Single-molecule speckle (SiMS) microscopy is a powerful method to directly elucidate biochemical reactions in live cells. However, since the signal from an individual fluorophore is extremely faint, the observation area by epi-fluorescence microscopy is restricted to the thin cell periphery to reduce autofluorescence, or only molecules near the plasma membrane are visualized by total internal reflection fluorescence (TIRF) microscopy. Here, we introduce a new actin probe labeled with near infrared (NIR) emissive CF680R dye for easy-to-use, electroporation-based SiMS microscopy (eSiMS) for deep-cell observation. CF680R-labeled actin (CF680R-actin) incorporated into actin structures and showed excellent brightness and photostability suitable for single-molecule imaging. Importantly, the intensity of autofluorescence with respect to SiMS brightness was reduced to approximately 13% compared to DyLight 550-labeled actin (DL550-actin). CF680R-actin enabled the monitoring of actin SiMS in actomyosin bundles associated with adherens junctions (AJs) located at 3.5–4 µm above the basal surfaces of epithelial monolayers. These favorable properties of CF680R-actin extend the application of eSiMS to actin turnover and flow analyses in deep cellular structures. PMID:28671584
Analysis and Modeling of Chromosome Congression During Mitosis in the Chemotherapy Drug Cisplatin.
Chacón, Jeremy M; Gardner, Melissa K
2013-12-01
The chemotherapy drug Cisplatin (cis-diamminedichloroplatinum(II)) induces crosslinks within and between DNA strands, and between DNA and nearby proteins. Therefore, Cisplatin-treated cells which progress into cell division may do so with altered chromosome mechanical properties. This could have important consequences for the successful completion of mitosis. Using Total Internal Reflection Fluorescence (TIRF) microscopy of live Cisplatin-treated Saccharomyces cerevisiae cells, we found that metaphase mitotic spindles have disorganized kinetochores relative to untreated cells, and also that there is increased variability in the chromosome stretching distance between sister centromeres. This suggests that chromosome stiffness may become more variable after Cisplatin treatment. We explored the effect of variable chromosome stiffness during mitosis using a stochastic model in which kinetochore microtubule dynamics were regulated by tension imparted by stretched sister chromosomes. Consistent with experimental results, increased variability of chromosome stiffness in the model led to disorganization of kinetochores in simulated metaphase mitotic spindles. Furthermore, the variability in simulated chromosome stretching tension was increased as chromosome stiffness became more variable. Because proper chromosome stretching tension may serve as a signal that is required for proper progression through mitosis, tension variability could act to impair this signal and thus prevent proper mitotic progression. Our results suggest a possible mitotic mode of action for the anti-cancer drug Cisplatin.
Adenylyl cyclase localization to the uropod of aggregating Dictyostelium cells requires RacC
Wang, C.; Jung, D.; Cao, Z.; Chung, C. Y.
2015-01-01
The localization of adenylyl cyclase A (ACA) to uropod of cells is required for the stream formation during Dictyostelium development. RacC is a Dictyostelium orthologue of Cdc42. We identified a streaming defect of racC− cells as they are clearly less polarized and form smaller and fragmented streams. ACA-YFP is mainly associated with intracellular vesicular structures, but not with the plasma membrane in racC− cells. racC− cells have a slightly higher number of vesicles than Ax3 cells, suggesting that the defect of ACA trafficking is not simply due to the lack of vesicle formation. While the ACA-YFP vesicles traveled with an average velocity of 9.1 µm/min in Ax3 cells, a slow and diffusional movement without direction with an average velocity of 4 µm/min was maintained in racC− cells. Images acquired by using total internal reflection fluorescence (TIRF) microscopy and fluorescence recovery after photobleaching (FRAP) analysis revealed that a significantly decreased number of ACA-YFP vesicles appeared near the cell membrane, indicating a defect in ACA-YFP vesicle trafficking. These results suggest an important role of RacC in the rapid and directional movements of ACA vesicles on microtubules to the plasma membrane, especially to the back of polarized cell. PMID:26315268
Interaction with Phosphoinositides Confers Adaptation onto the TRPV1 Pain Receptor
Yao, Jing; Qin, Feng
2009-01-01
Adaptation is a common feature of many sensory systems. But its occurrence to pain sensation has remained elusive. Here we address the problem at the receptor level and show that the capsaicin ion channel TRPV1, which mediates nociception at the peripheral nerve terminals, possesses properties essential to the adaptation of sensory responses. Ca2+ influx following the channel opening caused a profound shift (∼14-fold) of the agonist sensitivity, but did not alter the maximum attainable current. The shift was adequate to render the channel irresponsive to normally saturating concentrations, leaving the notion that the channel became no longer functional after desensitization. By simultaneous patch-clamp recording and total internal reflection fluorescence (TIRF) imaging, it was shown that the depletion of phosphatidylinositol 4,5-bisphosphate (PIP2) induced by Ca2+ influx had a rapid time course synchronous to the desensitization of the current. The extent of the depletion was comparable to that by rapamycin-induced activation of a PIP2 5-phosphatase, which also caused a significant reduction of the agonist sensitivity without affecting the maximum response. These results support a prominent contribution of PIP2 depletion to the desensitization of TRPV1 and suggest the adaptation as a possible physiological function for the Ca2+ influx through the channel. PMID:19243225
Basset, Antoine; Bouthemy, Patrick; Boulanger, Jérôme; Waharte, François; Salamero, Jean; Kervrann, Charles
2017-07-24
Characterizing membrane dynamics is a key issue to understand cell exchanges with the extra-cellular medium. Total internal reflection fluorescence microscopy (TIRFM) is well suited to focus on the late steps of exocytosis at the plasma membrane. However, it is still a challenging task to quantify (lateral) diffusion and estimate local dynamics of proteins. A new model was introduced to represent the behavior of cargo transmembrane proteins during the vesicle fusion to the plasma membrane at the end of the exocytosis process. Two biophysical parameters, the diffusion coefficient and the release rate parameter, are automatically estimated from TIRFM image sequences, to account for both the lateral diffusion of molecules at the membrane and the continuous release of the proteins from the vesicle to the plasma membrane. Quantitative evaluation on 300 realistic computer-generated image sequences demonstrated the efficiency and accuracy of the method. The application of our method on 16 real TIRFM image sequences additionally revealed differences in the dynamic behavior of Transferrin Receptor (TfR) and Langerin proteins. An automated method has been designed to simultaneously estimate the diffusion coefficient and the release rate for each individual vesicle fusion event at the plasma membrane in TIRFM image sequences. It can be exploited for further deciphering cell membrane dynamics.
Assembly of RNA nanostructures on supported lipid bilayers
NASA Astrophysics Data System (ADS)
Dabkowska, Aleksandra P.; Michanek, Agnes; Jaeger, Luc; Rabe, Michael; Chworos, Arkadiusz; Höök, Fredrik; Nylander, Tommy; Sparr, Emma
2014-12-01
The assembly of nucleic acid nanostructures with controlled size and shape has large impact in the fields of nanotechnology, nanomedicine and synthetic biology. The directed arrangement of nano-structures at interfaces is important for many applications. In spite of this, the use of laterally mobile lipid bilayers to control RNA three-dimensional nanostructure formation on surfaces remains largely unexplored. Here, we direct the self-assembly of RNA building blocks into three-dimensional structures of RNA on fluid lipid bilayers composed of cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or mixtures of zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) and cationic sphingosine. We demonstrate the stepwise supramolecular assembly of discrete building blocks through specific and selective RNA-RNA interactions, based on results from quartz crystal microbalance with dissipation (QCM-D), ellipsometry, fluorescence recovery after photobleaching (FRAP) and total internal reflection fluorescence microscopy (TIRF) experiments. The assembly can be controlled to give a densely packed single layer of RNA polyhedrons at the fluid lipid bilayer surface. We show that assembly of the 3D structure can be modulated by sequence specific interactions, surface charge and changes in the salt composition and concentration. In addition, the tertiary structure of the RNA polyhedron can be controllably switched from an extended structure to one that is dense and compact. The versatile approach to building up three-dimensional structures of RNA does not require modification of the surface or the RNA molecules, and can be used as a bottom-up means of nanofabrication of functionalized bio-mimicking surfaces.The assembly of nucleic acid nanostructures with controlled size and shape has large impact in the fields of nanotechnology, nanomedicine and synthetic biology. The directed arrangement of nano-structures at interfaces is important for many applications. In spite of this, the use of laterally mobile lipid bilayers to control RNA three-dimensional nanostructure formation on surfaces remains largely unexplored. Here, we direct the self-assembly of RNA building blocks into three-dimensional structures of RNA on fluid lipid bilayers composed of cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or mixtures of zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) and cationic sphingosine. We demonstrate the stepwise supramolecular assembly of discrete building blocks through specific and selective RNA-RNA interactions, based on results from quartz crystal microbalance with dissipation (QCM-D), ellipsometry, fluorescence recovery after photobleaching (FRAP) and total internal reflection fluorescence microscopy (TIRF) experiments. The assembly can be controlled to give a densely packed single layer of RNA polyhedrons at the fluid lipid bilayer surface. We show that assembly of the 3D structure can be modulated by sequence specific interactions, surface charge and changes in the salt composition and concentration. In addition, the tertiary structure of the RNA polyhedron can be controllably switched from an extended structure to one that is dense and compact. The versatile approach to building up three-dimensional structures of RNA does not require modification of the surface or the RNA molecules, and can be used as a bottom-up means of nanofabrication of functionalized bio-mimicking surfaces. Electronic supplementary information (ESI) available: Table with sequences of tRNA units used in this study; schematic structures of the RNA polyhedron and its building blocks; gel electrophoresis characterization of the RNA polyhedron and squares; AFM characterization of RNA tectosquare; schematic structures of RNA-9 and RNA-10 and their association with lipid bilayers; QCM-D frequency and dissipation data (as function of time) for adsorption of RNA polyhedrons, RNA squares and RNA9-10 TIRF images of RNA with Gelstar after photobleaching with analysis; Correlation plot in change of shear viscosity for TS3 and TO3-4 models for the stoichiometry of TS; QCM-D dissipation data for the sequential experiment in Fig. 5a; QCM-D and for the assembly of building blocks at the bilayer scaffold at varying bulk concentrations; QCM-D of adsorption of TS3. See DOI: 10.1039/c4nr05968a
The development of optical microscopy techniques for the advancement of single-particle studies
DOE Office of Scientific and Technical Information (OSTI.GOV)
Marchuk, Kyle
2013-05-15
Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-fieldmore » imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called “non-blinking” quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to find the 3D orientation of stationary metallic anisotropic nanoparticles utilizing only long-axis SPR enhancement. The polarization direction of the illuminating light was rotated causing the relative intensity of p-polarized and s-polarized light within the evanescent field to change. The interaction of the evanescent field with the particles is dependent on the orientation of the particle producing an intensity curve. This curve and the in-plane angle can be compared with simulations to accurately determine the 3D orientation. Differential interference contrast (DIC) microscopy is another non-invasive far-field technique based upon interferometry that does not rely on staining or other contrast enhancing techniques. In addition, high numerical aperture condensers and objectives can be used to give a very narrow depth of field allowing for the optical tomography of samples, which makes it an ideal candidate to study biological systems. DIC microscopy has also proven itself in determining the orientation of gold nanorods in both engineered environments and within cells. Many types of nanoparticles and nanostructures have been synthesized using lithographic techniques on silicon wafer substrates. Traditionally, reflective mode DIC microscopes have been developed and applied to the topographical study of reflective substrates and the imaging of chips on silicon wafers. Herein, a laser-illuminated reflected-mode DIC was developed for studying nanoparticles on reflective surfaces.« less
The development of optical microscopy techniques for the advancement of single-particle studies
NASA Astrophysics Data System (ADS)
Marchuk, Kyle
Single particle orientation and rotational tracking (SPORT) has recently become a powerful optical microscopy tool that can expose many molecular motions. Unfortunately, there is not yet a single microscopy technique that can decipher all particle motions in all environmental conditions, thus there are limitations to current technologies. Within, the two powerful microscopy tools of total internal reflection and interferometry are advanced to determine the position, orientation, and optical properties of metallic nanoparticles in a variety of environments. Total internal reflection is an optical phenomenon that has been applied to microscopy to produce either fluorescent or scattered light. The non-invasive far-field imaging technique is coupled with a near-field illumination scheme that allows for better axial resolution than confocal microscopy and epi-fluorescence microscopy. By controlling the incident illumination angle using total internal reflection fluorescence (TIRF) microscopy, a new type of imaging probe called "non-blinking" quantum dots (NBQDs) were super-localized in the axial direction to sub-10-nm precision. These particles were also used to study the rotational motion of microtubules being propelled by the motor protein kinesin across the substrate surface. The same instrument was modified to function under total internal reflection scattering (TIRS) microscopy to study metallic anisotropic nanoparticles and their dynamic interactions with synthetic lipid bilayers. Utilizing two illumination lasers with opposite polarization directions at wavelengths corresponding to the short and long axis surface plasmon resonance (SPR) of the nanoparticles, both the in-plane and out-of-plane movements of many particles could be tracked simultaneously. When combined with Gaussian point spread function (PSF) fitting for particle super-localization, the binding status and rotational movement could be resolved without degeneracy. TIRS microscopy was also used to find the 3D orientation of stationary metallic anisotropic nanoparticles utilizing only long-axis SPR enhancement. The polarization direction of the illuminating light was rotated causing the relative intensity of p-polarized and s-polarized light within the evanescent field to change. The interaction of the evanescent field with the particles is dependent on the orientation of the particle producing an intensity curve. This curve and the in-plane angle can be compared with simulations to accurately determine the 3D orientation. Differential interference contrast (DIC) microscopy is another non-invasive far-field technique based upon interferometry that does not rely on staining or other contrast enhancing techniques. In addition, high numerical aperture condensers and objectives can be used to give a very narrow depth of field allowing for the optical tomography of samples, which makes it an ideal candidate to study biological systems. DIC microscopy has also proven itself in determining the orientation of gold nanorods in both engineered environments and within cells. Many types of nanoparticles and nanostructures have been synthesized using lithographic techniques on silicon wafer substrates. Traditionally, reflective mode DIC microscopes have been developed and applied to the topographical study of reflective substrates and the imaging of chips on silicon wafers. Herein, a laser-illuminated reflected-mode DIC was developed for studying nanoparticles on reflective surfaces.
Pahlavan, Sara; Morad, Marin
2017-09-01
The details of cardiac Ca 2+ signaling within the dyadic junction remain unclear because of limitations in rapid spatial imaging techniques, and availability of Ca 2+ probes localized to dyadic junctions. To critically monitor ryanodine receptors' (RyR2) Ca 2+ nano-domains, we combined the use of genetically engineered RyR2-targeted pericam probes, (FKBP-YCaMP, K d =150nM, or FKBP-GCaMP6, K d =240nM) with rapid total internal reflectance fluorescence (TIRF) microscopy (resolution, ∼80nm). The punctate z-line patterns of FKBP, 2 -targeted probes overlapped those of RyR2 antibodies and sharply contrasted to the images of probes targeted to sarcoplasmic reticulum (SERCA2a/PLB), or cytosolic Fluo-4 images. FKBP-YCaMP signals were too small (∼20%) and too slow (2-3s) to detect Ca 2+ sparks, but the probe was effective in marking where Fluo-4 Ca 2+ sparks developed. FKBP-GCaMP6, on the other hand, produced rapidly decaying Ca 2+ signals that: a) had faster kinetics and activated synchronous with I Ca 3 but were of variable size at different z-lines and b) were accompanied by spatially confined spontaneous Ca 2+ sparks, originating from a subset of eager sites. The frequency of spontaneously occurring sparks was lower in FKBP-GCaMP6 infected myocytes as compared to Fluo-4 dialyzed myocytes, but isoproterenol enhanced their frequency more effectively than in Fluo-4 dialyzed cells. Nevertheless, isoproterenol failed to dissociate FKBP-GCaMP6 from the z-lines. The data suggests that FKBP-GCaMP6 binds predominantly to junctional RyR2s and has sufficient on-rate efficiency as to monitor the released Ca 2+ in individual dyadic clefts, and supports the idea that β-adrenergic agonists may modulate the stabilizing effects of native FKBP on RyR2. Copyright © 2017 Elsevier Ltd. All rights reserved.
AKAP150-mediated TRPV1 sensitization is disrupted by calcium/calmodulin
2011-01-01
Background The transient receptor potential vanilloid type1 (TRPV1) is expressed in nociceptive sensory neurons and is sensitive to phosphorylation. A-Kinase Anchoring Protein 79/150 (AKAP150) mediates phosphorylation of TRPV1 by Protein Kinases A and C, modulating channel activity. However, few studies have focused on the regulatory mechanisms that control AKAP150 association with TRPV1. In the present study, we identify a role for calcium/calmodulin in controlling AKAP150 association with, and sensitization of, TRPV1. Results In trigeminal neurons, intracellular accumulation of calcium reduced AKAP150 association with TRPV1 in a manner sensitive to calmodulin antagonism. This was also observed in transfected Chinese hamster ovary (CHO) cells, providing a model for conducting molecular analysis of the association. In CHO cells, the deletion of the C-terminal calmodulin-binding site of TRPV1 resulted in greater association with AKAP150, and increased channel activity. Furthermore, the co-expression of wild-type calmodulin in CHOs significantly reduced TRPV1 association with AKAP150, as evidenced by total internal reflective fluorescence-fluorescence resonance energy transfer (TIRF-FRET) analysis and electrophysiology. Finally, dominant-negative calmodulin co-expression increased TRPV1 association with AKAP150 and increased basal and PKA-sensitized channel activity. Conclusions the results from these studies indicate that calcium/calmodulin interferes with the association of AKAP150 with TRPV1, potentially extending resensitization of the channel. PMID:21569553
STIM1L traps and gates Orai1 channels without remodeling the cortical ER
Saüc, Sophie; Bulla, Monica; Nunes, Paula; Orci, Lelio; Marchetti, Anna; Antigny, Fabrice; Bernheim, Laurent; Cosson, Pierre; Frieden, Maud; Demaurex, Nicolas
2015-01-01
STIM proteins populate and expand cortical endoplasmic reticulum (ER) sheets to mediate store-operated Ca2+ entry (SOCE) by trapping and gating Orai channels in ER-plasma membrane clusters. A longer splice variant, STIM1L, forms permanent ER-plasma membrane clusters and mediates rapid Ca2+ influx in muscle. Here, we used electron microscopy, total internal reflection fluorescence (TIRF) microscopy and Ca2+ imaging to establish the trafficking and signaling properties of the two STIM1 isoforms in Stim1−/−/Stim2−/− fibroblasts. Unlike STIM1, STIM1L was poorly recruited into ER-plasma membrane clusters and did not mediate store-dependent expansion of cortical ER cisternae. Removal of the STIM1 lysine-rich tail prevented store-dependent cluster enlargement, whereas inhibition of cytosolic Ca2+ elevations or removal of the STIM1L actin-binding domain had no impact on cluster expansion. Finally, STIM1L restored robust but not accelerated SOCE and clustered with Orai1 channels more slowly than STIM1 following store depletion. These results indicate that STIM1L does not mediate rapid SOCE but can trap and gate Orai1 channels efficiently without remodeling cortical ER cisternae. The ability of STIM proteins to induce cortical ER formation is dispensable for SOCE and requires the lysine-rich tail of STIM1 involved in binding to phosphoinositides. PMID:25736291
AKAP150-mediated TRPV1 sensitization is disrupted by calcium/calmodulin.
Chaudhury, Sraboni; Bal, Manjot; Belugin, Sergei; Shapiro, Mark S; Jeske, Nathaniel A
2011-05-14
The transient receptor potential vanilloid type1 (TRPV1) is expressed in nociceptive sensory neurons and is sensitive to phosphorylation. A-Kinase Anchoring Protein 79/150 (AKAP150) mediates phosphorylation of TRPV1 by Protein Kinases A and C, modulating channel activity. However, few studies have focused on the regulatory mechanisms that control AKAP150 association with TRPV1. In the present study, we identify a role for calcium/calmodulin in controlling AKAP150 association with, and sensitization of, TRPV1. In trigeminal neurons, intracellular accumulation of calcium reduced AKAP150 association with TRPV1 in a manner sensitive to calmodulin antagonism. This was also observed in transfected Chinese hamster ovary (CHO) cells, providing a model for conducting molecular analysis of the association. In CHO cells, the deletion of the C-terminal calmodulin-binding site of TRPV1 resulted in greater association with AKAP150, and increased channel activity. Furthermore, the co-expression of wild-type calmodulin in CHOs significantly reduced TRPV1 association with AKAP150, as evidenced by total internal reflective fluorescence-fluorescence resonance energy transfer (TIRF-FRET) analysis and electrophysiology. Finally, dominant-negative calmodulin co-expression increased TRPV1 association with AKAP150 and increased basal and PKA-sensitized channel activity. the results from these studies indicate that calcium/calmodulin interferes with the association of AKAP150 with TRPV1, potentially extending resensitization of the channel.
The Intracellular Trafficking Pathway of Transferrin
Mayle, Kristine M.; Le, Alexander M.; Kamei, Daniel T.
2011-01-01
Background Transferrin (Tf) is an iron-binding protein that facilitates iron-uptake in cells. Iron-loaded Tf first binds to the Tf receptor (TfR) and enters the cell through clathrin-mediated endocytosis. Inside the cell, Tf is trafficked to early endosomes, delivers iron, and then is subsequently directed to recycling endosomes to be taken back to the cell surface. Scope of Review We aim to review the various methods and techniques that researchers have employed for elucidating the Tf trafficking pathway and the cell-machinery components involved. These experimental methods can be categorized as microscopy, radioactivity, and surface plasmon resonance (SPR). Major Conclusions Qualitative experiments, such as total internal reflectance fluorescence (TIRF), electron, laser-scanning confocal, and spinning-disk confocal microscopy, have been utilized to determine the roles of key components in the Tf trafficking pathway. These techniques allow temporal resolution and are useful for imaging Tf endocytosis and recycling, which occur on the order of seconds to minutes. Additionally, radiolabeling and SPR methods, when combined with mathematical modeling, have enabled researchers to estimate quantitative kinetic parameters and equilibrium constants associated with Tf binding and trafficking. General Significance Both qualitative and quantitative data can be used to analyze the Tf trafficking pathway. The valuable information that is obtained about the Tf trafficking pathway can then be combined with mathematical models to identify design criteria to improve the ability of Tf to deliver anticancer drugs. PMID:21968002
Myosin VI facilitates connexin 43 gap junction accretion.
Waxse, Bennett J; Sengupta, Prabuddha; Hesketh, Geoffrey G; Lippincott-Schwartz, Jennifer; Buss, Folma
2017-03-01
In this study, we demonstrate myosin VI enrichment at Cx43 (also known as GJA1)-containing gap junctions (GJs) in heart tissue, primary cardiomyocytes and cell culture models. In primary cardiac tissue and in fibroblasts from the myosin VI-null mouse as well as in tissue culture cells transfected with siRNA against myosin VI, we observe reduced GJ plaque size with a concomitant reduction in intercellular communication, as shown by fluorescence recovery after photobleaching (FRAP) and a new method of selective calcein administration. Analysis of the molecular role of myosin VI in Cx43 trafficking indicates that myosin VI is dispensable for the delivery of Cx43 to the cell surface and connexon movement in the plasma membrane. Furthermore, we cannot corroborate clathrin or Dab2 localization at gap junctions and we do not observe a function for the myosin-VI-Dab2 complex in clathrin-dependent endocytosis of annular gap junctions. Instead, we found that myosin VI was localized at the edge of Cx43 plaques by using total internal reflection fluorescence (TIRF) microscopy and use FRAP to identify a plaque accretion defect as the primary manifestation of myosin VI loss in Cx43 homeostasis. A fuller understanding of this derangement may explain the cardiomyopathy or gliosis associated with the loss of myosin VI. © 2017. Published by The Company of Biologists Ltd.
Pinaud, Fabien; Michalet, Xavier; Iyer, Gopal; Margeat, Emmanuel; Moore, Hsiao-Ping; Weiss, Shimon
2009-01-01
Recent experimental developments have led to a revision of the classical fluid mosaic model proposed by Singer and Nicholson 35 years ago. In particular, it is now well established that lipids and proteins diffuse heterogeneously in cell plasma membranes. Their complex motion patterns reflect the dynamic structure and composition of the membrane itself, as well as the presence of the underlying cytoskeleton scaffold and that of the extracellular matrix. How the structural organization of plasma membranes influences the diffusion of individual proteins remains a challenging, yet central question for cell signaling and its regulation. Here we have developed a raft-associated glycosylphosphatidyl Inositol-anchored avidin test probe (Av-GPI), whose diffusion patterns indirectly reports on the structure and dynamics of putative raft microdomains in the membrane of HeLa cells. Labeling with quantum dots (qdots) allowed high-resolution and long-term tracking of individual Av-GPI and the classification of their various diffusive behaviors. Using dual-color total internal reflection fluorescence (TIRF) microscopy, we studied the correlation between the diffusion of individual Av-GPI and the location of glycosphingolipid GM1-rich microdomains and caveolae. We show that Av-GPI exhibit a fast and a slow diffusion regime in different membrane regions, and that slowing down of their diffusion is correlated with entry in GM1-rich microdomains located in close proximity to, but distinct, from caveolae. We further show that Av-GPI dynamically partition in and out of these microdomains in a cholesterol-dependent manner. Our results provide direct evidence that cholesterol/sphingolipid-rich microdomains can compartmentalize the diffusion of GPI-anchored proteins in living cells and that the dynamic partitioning raft model appropriately describes the diffusive behavior of some raft-associated proteins across the plasma membrane. PMID:19416475
Pinaud, Fabien; Michalet, Xavier; Iyer, Gopal; Margeat, Emmanuel; Moore, Hsiao-Ping; Weiss, Shimon
2009-06-01
Recent experimental developments have led to a revision of the classical fluid mosaic model proposed by Singer and Nicholson more than 35 years ago. In particular, it is now well established that lipids and proteins diffuse heterogeneously in cell plasma membranes. Their complex motion patterns reflect the dynamic structure and composition of the membrane itself, as well as the presence of the underlying cytoskeleton scaffold and that of the extracellular matrix. How the structural organization of plasma membranes influences the diffusion of individual proteins remains a challenging, yet central, question for cell signaling and its regulation. Here we have developed a raft-associated glycosyl-phosphatidyl-inositol-anchored avidin test probe (Av-GPI), whose diffusion patterns indirectly report on the structure and dynamics of putative raft microdomains in the membrane of HeLa cells. Labeling with quantum dots (qdots) allowed high-resolution and long-term tracking of individual Av-GPI and the classification of their various diffusive behaviors. Using dual-color total internal reflection fluorescence (TIRF) microscopy, we studied the correlation between the diffusion of individual Av-GPI and the location of glycosphingolipid GM1-rich microdomains and caveolae. We show that Av-GPI exhibit a fast and a slow diffusion regime in different membrane regions, and that slowing down of their diffusion is correlated with entry in GM1-rich microdomains located in close proximity to, but distinct, from caveolae. We further show that Av-GPI dynamically partition in and out of these microdomains in a cholesterol-dependent manner. Our results provide direct evidence that cholesterol-/sphingolipid-rich microdomains can compartmentalize the diffusion of GPI-anchored proteins in living cells and that the dynamic partitioning raft model appropriately describes the diffusive behavior of some raft-associated proteins across the plasma membrane.
Zarcone, Jennifer; Hagopian, Louis; Ninci, Jennifer; McKay, Chloe; Bonner, Andrew; Dillon, Christopher; Hausman, Nicole
2016-01-01
Objectives The goal of this study was to develop and evaluate a tool to measure the complexity and intensity of psychotropic medication interventions, behavioral interventions, and issues related to crisis management for challenging behavior using a standardized rating form. Method The Treatment Intensity Rating Form (TIRF) is a 10-item scale with three categories: pharmacological interventions, behavior supports, and protective equipment. In a retrospective review we examined the final treatment recommendations for 74 individuals with self-injurious behavior (SIB) based on psychiatric and behavioral notes and reports. We also compared whether TIRF scores differed across individuals for whom SIB was maintained by social reinforcement (e.g., to access attention or toys/activities, or escape from tasks) versus those for whom SIB was maintained by automatic reinforcement (e.g., occurs independent of social variables, and is presumed to be maintained by sensory reinforcement). Results The TIRF was demonstrated to have strong inter-rater reliability (98%) and appears to have good face validity. As hypothesized, individuals with SIB maintained by automatic reinforcement had significantly more medication trials (p=0.0005) and required more protective equipment than individuals with SIB maintained by social reinforcement (p=0.0002). Antidepressant medication was used more often with individuals with automatically reinforced SIB, although antipsychotics and anticonvulsants were also commonly used across both groups. Conclusion Findings provide initial support for the TIRF’s reliability, and face validity as a measure the level of complexity of medical and behavioral treatment plans - although additional research is needed to fully evaluate its psychometric properties. PMID:27917287
Highly Permeable Silicon Membranes for Shear Free Chemotaxis and Rapid Cell Labeling
Chung, Henry H.; Chan, Charles K.; Khire, Tejas S.; Marsh, Graham A.; Clark, Alfred; Waugh, Richard E.; McGrath, James L.
2015-01-01
Microfluidic systems are powerful tools for cell biology studies because they enable the precise addition and removal of solutes in small volumes. However, the fluid forces inherent in the use of microfluidics for cell cultures are sometimes undesirable. An important example is chemotaxis systems where fluid flow creates well-defined and steady chemotactic gradients but also pushes cells downstream. Here we demonstrate a chemotaxis system in which two chambers are separated by a molecularly thin (15 nm), transparent, and nanoporous silicon membrane. One chamber is a microfluidic channel that carries a flow-generated gradient while the other chamber is a shear-free environment for cell observation. The molecularly thin membranes provide effectively no resistance to molecular diffusion between the two chambers, making them ideal elements for creating flow-free chambers in microfluidic systems. Analytical and computational flow models that account for membrane and chamber geometry, predict shear reduction of more than five orders of magnitude. This prediction is confirmed by observing the pure diffusion of nanoparticles in the cell-hosting chamber despite high input flow (Q = 10 µL min−1; vavg ~45 mm min−1) in the flow chamber only 15 nm away. Using total internal reflection fluorescence (TIRF) microscopy, we show that a flow-generated molecular gradient will pass through the membrane into the quiescent cell chamber. Finally we demonstrate that our device allows us to expose migrating neutrophils to a chemotactic gradient or fluorescent label without any influence from flow. PMID:24850320
Human Growth Hormone Adsorption Kinetics and Conformation on Self-Assembled Monolayers
Buijs, Jos; Britt, David W.; Hlady, Vladimir
2012-01-01
The adsorption process of the recombinant human growth hormone on organic films, created by self-assembly of octadecyltrichlorosilane, arachidic acid, and dipalmitoylphosphatidylcholine, is investigated and compared to adsorption on silica and methylated silica substrates. Information on the adsorption process of human growth hormone (hGH) is obtained by using total internal reflection fluorescence (TIRF). The intensity, spectra, and quenching of the intrinsic fluorescence emitted by the growth hormone’s single tryptophan are monitored and related to adsorption kinetics and protein conformation. For the various alkylated hydrophobic surfaces with differences in surface density and conformational freedom it is observed that the adsorbed amount of growth hormone is relatively large if the alkyl chains are in an ordered structure while the amounts adsorbed are considerably lower for adsorption onto less ordered alkyl chains of fatty acid and phospholipid layers. Adsorption on methylated surfaces results in a relatively large conformational change in the growth hormone’s structure, as displayed by a 7 nm blue shift in emission wavelength and a large increase in the effectiveness of fluorescence quenching. Conformational changes are less evident for hGH adsorption onto the fatty acid and phospholipid alkyl chains. Adsorption kinetics on the hydrophilic head groups of the self-assembled monolayers are similar to those on solid hydrophilic surfaces. The relatively small conformational changes in the hGH structure observed for adsorption on silica are even further reduced for adsorption on fatty acid head groups. PMID:25125795
Segmentation and Tracking of Cytoskeletal Filaments Using Open Active Contours
Smith, Matthew B.; Li, Hongsheng; Shen, Tian; Huang, Xiaolei; Yusuf, Eddy; Vavylonis, Dimitrios
2010-01-01
We use open active contours to quantify cytoskeletal structures imaged by fluorescence microscopy in two and three dimensions. We developed an interactive software tool for segmentation, tracking, and visualization of individual fibers. Open active contours are parametric curves that deform to minimize the sum of an external energy derived from the image and an internal bending and stretching energy. The external energy generates (i) forces that attract the contour toward the central bright line of a filament in the image, and (ii) forces that stretch the active contour toward the ends of bright ridges. Images of simulated semiflexible polymers with known bending and torsional rigidity are analyzed to validate the method. We apply our methods to quantify the conformations and dynamics of actin in two examples: actin filaments imaged by TIRF microscopy in vitro, and actin cables in fission yeast imaged by spinning disk confocal microscopy. PMID:20814909
Imaging Local Ca2+ Signals in Cultured Mammalian Cells
Lock, Jeffrey T.; Ellefsen, Kyle L.; Settle, Bret; Parker, Ian; Smith, Ian F.
2015-01-01
Cytosolic Ca2+ ions regulate numerous aspects of cellular activity in almost all cell types, controlling processes as wide-ranging as gene transcription, electrical excitability and cell proliferation. The diversity and specificity of Ca2+ signaling derives from mechanisms by which Ca2+ signals are generated to act over different time and spatial scales, ranging from cell-wide oscillations and waves occurring over the periods of minutes to local transient Ca2+ microdomains (Ca2+ puffs) lasting milliseconds. Recent advances in electron multiplied CCD (EMCCD) cameras now allow for imaging of local Ca2+ signals with a 128 x 128 pixel spatial resolution at rates of >500 frames sec-1 (fps). This approach is highly parallel and enables the simultaneous monitoring of hundreds of channels or puff sites in a single experiment. However, the vast amounts of data generated (ca. 1 Gb per min) render visual identification and analysis of local Ca2+ events impracticable. Here we describe and demonstrate the procedures for the acquisition, detection, and analysis of local IP3-mediated Ca2+ signals in intact mammalian cells loaded with Ca2+ indicators using both wide-field epi-fluorescence (WF) and total internal reflection fluorescence (TIRF) microscopy. Furthermore, we describe an algorithm developed within the open-source software environment Python that automates the identification and analysis of these local Ca2+ signals. The algorithm localizes sites of Ca2+ release with sub-pixel resolution; allows user review of data; and outputs time sequences of fluorescence ratio signals together with amplitude and kinetic data in an Excel-compatible table. PMID:25867132
Assembly of RNA nanostructures on supported lipid bilayers
Dabkowska, Aleksandra P.; Michanek, Agnes; Jaeger, Luc; Rabe, Michael; Chworos, Arkadiusz; Höök, Fredrik; Nylander, Tommy; Sparr, Emma
2014-01-01
The assembly of nucleic acid nanostructures with controlled size and shape has large impact in the fields of nanotechnology, nanomedicine and synthetic biology. The directed arrangement of nanostructures at interfaces is important for many applications. In spite of this, the use of laterally mobile lipid bilayers to control RNA three-dimensional nanostructure formation on surfaces remains largely unexplored. Here, we direct the self-assembly of RNA building blocks into three-dimensional structures of RNA on fluid lipid bilayers composed of cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or mixtures of zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) and cationic sphingosine. We demonstrate the stepwise supramolecular assembly of discrete building blocks through specific and selective RNA-RNA interactions, based on results from quartz crystal microbalance with dissipation (QCM-D), ellipsometry, fluorescence recovery after photobleaching (FRAP) and total internal reflection fluorescence microscopy (TIRF) experiments. The assembly can be controlled to give a densely packed single layer of RNA polyhedrons at the fluid lipid bilayer surface. We show that assembly of the 3D structure can be modulated by sequence specific interactions, surface charge and changes in the salt composition and concentration. In addition, the tertiary structure of the RNA polyhedron can be controllably switched from an extended structure to one that is dense and compact. The versatile approach to building up three-dimensional structures of RNA does not require modification of the surface or the RNA molecules, and can be used as a bottom-up means of nanofabrication of functionalized bio-mimicking surfaces. PMID:25417592
Rodriguez-Mozaz, Sara; de Alda, Maria J López; Barceló, Damià
2006-04-15
This work describes the application of an optical biosensor (RIver ANALyser, RIANA) to the simultaneous analysis of three relevant environmental organic pollutants, namely, the pesticides atrazine and isoproturon and the estrogen estrone, in real water samples. This biosensor is based on an indirect inhibition immunoassay which takes place at a chemically modified optical transducer chip. The spatially resolved modification of the transducer surface allows the simultaneous determination of selected target analytes by means of "total internal reflection fluorescence" (TIRF). The performance of the immunosensor method developed was evaluated against a well accepted traditional method based on solid-phase extraction followed by liquid chromatography-mass spectrometry (LC-MS). The chromatographic method was superior in terms of linearity, sensitivity and accuracy, and the biosensor method in terms of repeatability, speed, cost and automation. The application of both methods in parallel to determine the occurrence and removal of atrazine, isoproturon and estrone throughout the treatment process (sand filtration, ozonation, activated carbon filtration and chlorination) in a waterworks showed an overestimation of results in the case of the biosensor, which was partially attributed to matrix and cross-reactivity effects, in spite of the addition of ovalbumin to the sample to minimize matrix interferences. Based on the comparative performance of both techniques, the biosensor emerges as a suitable tool for fast, simple and automated screening of water pollutants without sample pretreatment. To the author's knowledge, this is the first description of the application of the biosensor RIANA in the multi-analyte configuration to the regular monitoring of pollutants in a waterworks.
Injuries related to off-road vehicles in Canada.
Vanlaar, Ward; McAteer, Heather; Brown, Steve; Crain, Jennifer; McFaull, Steven; Hing, Marisela Mainegra
2015-02-01
Off-road vehicles (ORVs; this includes snowmobiles, all-terrain vehicles or ATVs and dirt bikes) were once used primarily for work and travel. Such use remains common in Canada, although their recreational use has also gained popularity in recent years. An epidemiological injury profile of ORV users is important for better understanding injuries and their risk factors to help inform injury prevention initiatives. The Traffic Injury Research Foundation (TIRF) partnered with the Public Health Agency of Canada (PHAC) to analyze the epidemiology of ORV-related injuries. The primary aim was to assess crashes and injuries in Canada, including the extent of alcohol involvement. Secondly, the burden of injury among children and teen ORV drivers in Canada, as well as passengers, was investigated. Descriptive and inferential epidemiological statistics were generated using the following data sources: first, TIRF's National Fatality Database, which is a comprehensive, pan-Canadian, set of core data on all fatal motor vehicle crashes; second, TIRF's Serious Injury Database, which contains information on persons seriously injured in crashes; and, third, PHAC's Canadian Hospitals Injury Reporting and Prevention Program (CHIRPP), a surveillance system currently operating in the emergency departments of some pediatric and general hospitals across Canada. Exposure data have been used in the analyzes where available. Between 1990 and 2010, fatality rates increased among ATV and dirt bike operators. The fatality rate among snowmobilers declined during this period. Of particular concern, among fatally injured female ATV users, children aged 0-15 years comprised the highest proportion of any age group at 33.8%. Regarding alcohol use, among fatally injured snowmobile and ATV/dirt bike operators tested for alcohol, 66% and 55% tested positive, respectively. Alcohol involvement in adult ORV crashes remains an important factor. In light of the growing popularity of ORVs, prevention and mitigation measures are required to address this issue. Copyright © 2014 Elsevier Ltd. All rights reserved.
Nitroxide amide-BODIPY probe behavior in fibroblasts analyzed by advanced fluorescence microscopy.
Liras, M; Simoncelli, S; Rivas-Aravena, A; García, O; Scaiano, J C; Alarcon, E I; Aspée, A
2016-04-26
A novel synthesized nitroxide amide-BODIPY prefluorescent probe was used to study cellular redox balance that modulates nitroxide/hydroxylamine ratio in cultured human fibroblasts. FLIM quantitatively differentiated between nitroxide states of the cytoplasm-localized probe imaged by TIRF, monitoring nitroxide depletion by hydrogen peroxide; eluding incorrect interpretation if only fluorescence intensity is considered.
Developing new optical imaging techniques for single particle and molecule tracking in live cells
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sun, Wei
Differential interference contrast (DIC) microscopy is a far-field as well as wide-field optical imaging technique. Since it is non-invasive and requires no sample staining, DIC microscopy is suitable for tracking the motion of target molecules in live cells without interfering their functions. In addition, high numerical aperture objectives and condensers can be used in DIC microscopy. The depth of focus of DIC is shallow, which gives DIC much better optical sectioning ability than those of phase contrast and dark field microscopies. In this work, DIC was utilized to study dynamic biological processes including endocytosis and intracellular transport in live cells.more » The suitability of DIC microscopy for single particle tracking in live cells was first demonstrated by using DIC to monitor the entire endocytosis process of one mesoporous silica nanoparticle (MSN) into a live mammalian cell. By taking advantage of the optical sectioning ability of DIC, we recorded the depth profile of the MSN during the endocytosis process. The shape change around the nanoparticle due to the formation of a vesicle was also captured. DIC microscopy was further modified that the sample can be illuminated and imaged at two wavelengths simultaneously. By using the new technique, noble metal nanoparticles with different shapes and sizes were selectively imaged. Among all the examined metal nanoparticles, gold nanoparticles in rod shapes were found to be especially useful. Due to their anisotropic optical properties, gold nanorods showed as diffraction-limited spots with disproportionate bright and dark parts that are strongly dependent on their orientation in the 3D space. Gold nanorods were developed as orientation nanoprobes and were successfully used to report the self-rotation of gliding microtubules on kinesin coated substrates. Gold nanorods were further used to study the rotational motions of cargoes during the endocytosis and intracellular transport processes in live mammalian cells. New rotational information was obtained: (1) during endocytosis, cargoes lost their rotation freedom at the late stage of internalization; (2) cargoes performed train-like motion when they were transported along the microtubule network by motor proteins inside live cells; (3) During the pause stage of fast axonal transport, cargoes were still bound to the microtubule tracks by motor proteins. Total internal reflection fluorescence microscopy (TIRFM) is another non-invasive and far-field optical imaging technique. Because of its near-field illumination mechanism, TIRFM has better axial resolution than epi-fluorescence microscopy and confocal microscopy. In this work, an auto-calibrated, prism type, angle-scanning TIRFM instrument was built. The incident angle can range from subcritical angles to nearly 90°, with an angle interval less than 0.2°. The angle precision of the new instrument was demonstrated through the finding of the surface plasmon resonance (SPR) angle of metal film coated glass slide. The new instrument improved significantly the precision in determining the axial position. As a result, the best obtained axial resolution was ~ 8 nm, which is better than current existing instruments similar in function. The instrument was further modified to function as a pseudo TIRF microscope. The illumination depth can be controlled by changing the incident angle of the excitation laser beam or adjusting the horizontal position of the illumination laser spot on the prism top surface. With the new technique, i.e., variable-illumination-depth pseudo TIRF microscopy, the whole cell body from bottom to top was scanned.« less
ERIC Educational Resources Information Center
Page-Valin, L.; And Others
In its evaluation of the effectiveness of driver education as taught in secondary and commercial schools, this report (1) investigates the objectives of driver education, (2) determines what is presently known about its effectiveness through a review of previous evaluative research, (3) discusses what driver education is likely/unlikely to…
Single Molecule Visualization of Protein-DNA Complexes: Watching Machines at Work
NASA Astrophysics Data System (ADS)
Kowalczykowski, Stephen
2013-03-01
We can now watch individual proteins acting on single molecules of DNA. Such imaging provides unprecedented interrogation of fundamental biophysical processes. Visualization is achieved through the application of two complementary procedures. In one, single DNA molecules are attached to a polystyrene bead and are then captured by an optical trap. The DNA, a worm-like coil, is extended either by the force of solution flow in a micro-fabricated channel, or by capturing the opposite DNA end in a second optical trap. In the second procedure, DNA is attached by one end to a glass surface. The coiled DNA is elongated either by continuous solution flow or by subsequently tethering the opposite end to the surface. Protein action is visualized by fluorescent reporters: fluorescent dyes that bind double-stranded DNA (dsDNA), fluorescent biosensors for single-stranded DNA (ssDNA), or fluorescently-tagged proteins. Individual molecules are imaged using either epifluorescence microscopy or total internal reflection fluorescence (TIRF) microscopy. Using these approaches, we imaged the search for DNA sequence homology conducted by the RecA-ssDNA filament. The manner by which RecA protein finds a single homologous sequence in the genome had remained undefined for almost 30 years. Single-molecule imaging revealed that the search occurs through a mechanism termed ``intersegmental contact sampling,'' in which the randomly coiled structure of DNA is essential for reiterative sampling of DNA sequence identity: an example of parallel processing. In addition, the assembly of RecA filaments on single molecules of single-stranded DNA was visualized. Filament assembly requires nucleation of a protein dimer on DNA, and subsequent growth occurs via monomer addition. Furthermore, we discovered a class of proteins that catalyzed both nucleation and growth of filaments, revealing how the cell controls assembly of this protein-DNA complex.
Donahoe, Casey D.; Cohen, Thomas L.; Li, Wenlu; Nguyen, Peter K.; Fortner, John D.; Mitra, Robi D.; Elbert, Donald L.
2013-01-01
Clickable nanogel solutions were synthesized by using the copper catalyzed azide/alkyne cycloaddition (CuAAC) to partially polymerize solutions of azide and alkyne functionalized poly(ethylene glycol) (PEG) monomers. Coatings were fabricated using a second click reaction: a UV thiol-yne attachment of the nanogel solutions to mercaptosilanated glass. Because the CuAAC reaction was effectively halted by the addition of a copper-chelator, we were able to prevent bulk gelation and limit the coating thickness to a single monolayer of nanogels in the absence of the solution reaction. This enabled the inclusion of kosmotropic salts, which caused the PEG to phase-separate and nearly double the nanogel packing density, as confirmed by Quartz Crystal Microbalance with Dissipation (QCM-D). Protein adsorption was analyzed by single molecule counting with total internal reflection fluorescence (TIRF) microscopy and cell adhesion assays. Coatings formed from the phase-separated clickable nanogel solutions attached with salt adsorbed significantly less fibrinogen than other 100% PEG coatings tested, as well as poly-L-lysine-g-PEG (PLL-g-PEG) coatings. However, PEG/albumin nanogel coatings still outperformed the best 100% PEG clickable nanogel coatings. Additional surface crosslinking of the clickable nanogel coating in the presence of copper further reduced levels of fibrinogen adsorption closer to those of PEG/albumin nanogel coatings. However, this step negatively impacted long-term resistance to cell adhesion and dramatically altered the morphology of the coating by atomic force microscopy (AFM). The main benefit of the click strategy is that the partially polymerized solutions are stable almost indefinitely, allowing attachment in the phase-separated state without danger of bulk gelation, and thus, producing the best performing 100% PEG coating that we have studied to date. PMID:23441808
Ramabhadran, Vinay; Gurel, Pinar S.; Higgs, Henry N.
2012-01-01
INF2 (inverted formin 2) is a formin protein with unusual biochemical characteristics. As with other formins, the formin homology 2 (FH2) domain of INF2 accelerates actin filament assembly and remains at the barbed end, modulating elongation. The unique feature of INF2 is its ability to sever filaments and enhance depolymerization, which requires the C-terminal region. Physiologically, INF2 acts in the secretory pathway and is mutated in two human diseases, focal and segmental glomerulosclerosis and Charcot-Marie-Tooth disease. In this study, we investigate the effects of mutating two FH2 residues found to be key in other formins: Ile-643 and Lys-792. Surprisingly, neither mutation abolishes barbed end binding, as judged by pyrene-actin and total internal reflection (TIRF) microscopy elongation assays. The I643A mutation causes tight capping of a subset of filaments, whereas K792A causes slow elongation of all filaments. The I643A mutation has a minor inhibitory effect on polymerization activity but causes almost complete abolition of severing and depolymerization activity. The K792A mutation has relatively small effects on polymerization, severing, and depolymerization. In cells, the K792A mutant causes actin accumulation around the endoplasmic reticulum to a similar extent as wild type, whereas the I643A mutant causes no measurable polymerization. The inability of I643A to induce actin polymerization in cells is explained by its inability to promote robust actin polymerization in the presence of capping protein. These results highlight an important point: it is dangerous to assume that mutation of conserved FH2 residues will have equivalent effects in all formins. The work also suggests that both mutations have effects on the mechanism of processive elongation. PMID:22879592
von Einem, Bjoern; Weber, Petra; Wagner, Michael; Malnar, Martina; Kosicek, Marko; Hecimovic, Silva; Arnim, Christine A F von; Schneckenburger, Herbert
2012-11-26
Förster resonance energy transfer (FRET) -based techniques have recently been applied to study the interactions between β-site APP-cleaving enzyme-GFP (BACE1-GFP) and amyloid precursor protein-mRFP (APP-mRFP) in U373 glioblastoma cells. In this context, the role of APP-BACE1 proximity in Alzheimer's disease (AD) pathogenesis has been discussed. FRET was found to depend on intracellular cholesterol levels and associated alterations in membrane stiffness. Here, NPC1 null cells (CHO-NPC1-/-), exhibiting increased cholesterol levels and disturbed cholesterol transport similar to that observed in Niemann-Pick type C disease (NPC), were used to analyze the influence of altered cholesterol levels on APP-BACE1 proximity. Fluorescence lifetime measurements of whole CHO-wild type (WT) and CHO-NPC1-/- cells (EPI-illumination microscopy), as well as their plasma membranes (total internal reflection fluorescence microscopy, TIRFM), were performed. Additionally, generalized polarization (GP) measurements of CHO-WT and CHO-NPC1-/- cells incubated with the fluorescence marker laurdan were performed to determine membrane stiffness of plasma- and intracellular-membranes. CHO-NPC1-/- cells showed higher membrane stiffness at intracellular- but not plasma-membranes, equivalent to cholesterol accumulation in late endosomes/lysosomes. Along with higher membrane stiffness, the FRET efficiency between BACE1-GFP and APP-mRFP was reduced at intracellular membranes, but not within the plasma membrane of CHO-NPC1-/-. Our data show that FRET combined with TIRF is a powerful technique to determine protein proximity and membrane fluidity in cellular models of neurodegenerative diseases.
Vascular disease-causing mutation R258C in ACTA2 disrupts actin dynamics and interaction with myosin
Lu, Hailong; Fagnant, Patricia M.; Bookwalter, Carol S.; Joel, Peteranne; Trybus, Kathleen M.
2015-01-01
Point mutations in vascular smooth muscle α-actin (SM α-actin), encoded by the gene ACTA2, are the most prevalent cause of familial thoracic aortic aneurysms and dissections (TAAD). Here, we provide the first molecular characterization, to our knowledge, of the effect of the R258C mutation in SM α-actin, expressed with the baculovirus system. Smooth muscles are unique in that force generation requires both interaction of stable actin filaments with myosin and polymerization of actin in the subcortical region. Both aspects of R258C function therefore need investigation. Total internal reflection fluorescence (TIRF) microscopy was used to quantify the growth of single actin filaments as a function of time. R258C filaments are less stable than WT and more susceptible to severing by cofilin. Smooth muscle tropomyosin offers little protection from cofilin cleavage, unlike its effect on WT actin. Unexpectedly, profilin binds tighter to the R258C monomer, which will increase the pool of globular actin (G-actin). In an in vitro motility assay, smooth muscle myosin moves R258C filaments more slowly than WT, and the slowing is exacerbated by smooth muscle tropomyosin. Under loaded conditions, small ensembles of myosin are unable to produce force on R258C actin-tropomyosin filaments, suggesting that tropomyosin occupies an inhibitory position on actin. Many of the observed defects cannot be explained by a direct interaction with the mutated residue, and thus the mutation allosterically affects multiple regions of the monomer. Our results align with the hypothesis that defective contractile function contributes to the pathogenesis of TAAD. PMID:26153420
A-type granite and the Red Sea opening
Coleman, R.G.; DeBari, S.; Peterman, Z.
1992-01-01
Miocene-Oligocene A-type granite intrudes the eastern side of the Red Sea margin within the zone of extension from Jiddah, Saudi Arabia south to Yemen. The intrusions developed in the early stages of continental extension as Arabia began to move slowly away from Africa (around 30-20 Ma). Within the narrow zone of extension silicic magmas formed dikes, sills, small plutons and extrusive equivalents. In the Jabal Tirf area of Saudi Arabia these rocks occur in an elongate zone consisting of late Precambrian basement to the east, which is gradually invaded by mafic dikes. The number of dikes increases westward until an igneous complex is produced parallel to the present Red Sea axis. The Jabal Tirf igneous complex consists of diabase and rhyolite-granophyre sills (20-24 Ma). Although these are intrusine intrusive rocks their textures indicate shallow depths of intrusion (< 1 km). To the south, in the Yemen, contemporaneous with alkali basaltic eruptions (26-30 Ma) and later silicic eruptions, small plutons, dikes, and stocks of alkali granite invaded thick (1500 m) volcanic series, at various levels and times. Erosion within the uplifted margin of Yemen suggests that the maximum depth of intrusion was less than 1-2 km. Granophyric intrusions (20-30 Ma) within mafic dike swarms similar to the Jabal Tirf complex are present along the western edge of the Yemen volcanic plateau, marking a north-south zone of continental extension. The alkali granites of Yemen consist primarily of perthitic feldspar and quartz with some minor alkali amphiboles and acmite. These granites represent water-poor, hypersolvus magmas generated from parent alkali basalt magmas. The granophyric, two-feldspar granites associated with the mafic dike swarms and layered gabbros formed by fractional crystallization from tholeiitic basalt parent developed in the early stages of extension. Initial 87Sr/86Sr ratios of these rocks and their bulk chemistry indicate that production of peralkaline and metaluminous granitic magmas involved both fractio??nation and partial melting as they ascended through the late Precambrian crust of the Arabian plate. ?? 1992.
Visualization of self-delivering hydrophobically modified siRNA cellular internalization
Ly, Socheata; Navaroli, Deanna M.; Didiot, Marie-Cécile; Cardia, James; Pandarinathan, Lakshmipathi; Alterman, Julia F.; Fogarty, Kevin; Standley, Clive; Lifshitz, Lawrence M.; Bellve, Karl D.; Prot, Matthieu; Echeverria, Dimas; Corvera, Silvia; Khvorova, Anastasia
2017-01-01
siRNAs are a new class of therapeutic modalities with promising clinical efficacy that requires modification or formulation for delivery to the tissue and cell of interest. Conjugation of siRNAs to lipophilic groups supports efficient cellular uptake by a mechanism that is not well characterized. Here we study the mechanism of internalization of asymmetric, chemically stabilized, cholesterol-modified siRNAs (sd-rxRNAs®) that efficiently enter cells and tissues without the need for formulation. We demonstrate that uptake is rapid with significant membrane association within minutes of exposure followed by the formation of vesicular structures and internalization. Furthermore, sd-rxRNAs are internalized by a specific class of early endosomes and show preferential association with epidermal growth factor (EGF) but not transferrin (Tf) trafficking pathways as shown by live cell TIRF and structured illumination microscopy (SIM). In fixed cells, we observe ∼25% of sd-rxRNA co-localizing with EGF and <5% with Tf, which is indicative of selective endosomal sorting. Likewise, preferential sd-rxRNA co-localization was demonstrated with EEA1 but not RBSN-containing endosomes, consistent with preferential EGF-like trafficking through EEA1-containing endosomes. sd-rxRNA cellular uptake is a two-step process, with rapid membrane association followed by internalization through a selective, saturable subset of the endocytic process. However, the mechanistic role of EEA1 is not yet known. This method of visualization can be used to better understand the kinetics and mechanisms of hydrophobic siRNA cellular uptake and will assist in further optimization of these types of compounds for therapeutic intervention. PMID:27899655
DOE Office of Scientific and Technical Information (OSTI.GOV)
Nagamatsu, Shinya, E-mail: shinya@ks.kyorin-u.ac.jp; Ohara-Imaizumi, Mica; Nakamichi, Yoko
Highlights: {yields} Anti-diabetic new drug, DPP-4 inhibitor, can affect the insulin exocytosis. {yields} DPP-4 inhibitor treatment altered syntaxin 1 expression. {yields} Treatment of db/db mice with DPP-4 inhibitor increased insulin release. -- Abstract: Incretin promotes insulin secretion acutely. Recently, orally-administered DPP-4 inhibitors represent a new class of anti-hyperglycemic agents. Indeed, inhibitors of dipeptidyl peptidase-IV (DPP-4), sitagliptin, has just begun to be widely used as therapeutics for type 2 diabetes. However, the effects of sitagliptin-treatment on insulin exocytosis from single {beta}-cells are yet unknown. We therefore investigated how sitagliptin-treatment in db/db mice affects insulin exocytosis by treating db/db mice with des-F-sitagliptinmore » for 2 weeks. Perfusion studies showed that 2 weeks-sitagliptin treatment potentiated insulin secretion. We then analyzed insulin granule motion and SNARE protein, syntaxin 1, by TIRF imaging system. TIRF imaging of insulin exocytosis showed the increased number of docked insulin granules and increased fusion events from them during first-phase release. In accord with insulin exocytosis data, des-F-sitagliptin-treatment increased the number of syntaxin 1 clusters on the plasma membrane. Thus, our data demonstrated that 2-weeks des-F-sitagliptin-treatment increased the fusion events of insulin granules, probably via increased number of docked insulin granules and that of syntaxin 1 clusters.« less
NASA Astrophysics Data System (ADS)
Smith, Steve; Hor, Amy; Luu, Anh; Kang, Lin; Scott, Brandon; Bailey, Elizabeth; Hoppe, Adam
Clathrin-mediated endocytosis is one of the central pathways for cargo transport into cells, and plays a major role in the maintenance of cellular functions, such as intercellular signaling, nutrient intake, and turnover of plasma membrane in cells. The clathrin-mediated endocytosis process involves invagination and formation of clathrin-coated vesicles. However, the biophysical mechanisms of vesicle formation are still debated. We investigate clathrin vesicle formation mechanisms through the utilization of tapping-mode atomic force microscopy for high resolution topographical imaging in neutral buffer solution of unroofed cells exposing the inner membrane, combined with fluorescence imaging to definitively label intracellular constituents with specific fluorescent fusion proteins (actin filaments labeled with green phalloidin-antibody and clathrin coated vesicles with the fusion protein Tq2) in SKMEL (Human Melanoma) cells. Results from our work are compared against dynamical polarized total internal fluorescence (TIRF), super-resolution photo-activated localization microscopy (PALM) and transmission electron microscopy (TEM) to draw conclusions regarding the prominent model of vesicle formation in clathrin-mediated endocytosis. Funding provided by NSF MPS/DMR/BMAT award # 1206908.
Adsorption behavior of a human monoclonal antibody at hydrophilic and hydrophobic surfaces
Couston, Ruairidh G.; Skoda, Maximilian W.; Uddin, Shahid; van der Walle, Christopher F.
2013-01-01
One aspiration for the formulation of human monoclonal antibodies (mAb) is to reach high solution concentrations without compromising stability. Protein surface activity leading to instability is well known, but our understanding of mAb adsorption to the solid-liquid interface in relevant pH and surfactant conditions is incomplete. To investigate these conditions, we used total internal reflection fluorescence (TIRF) and neutron reflectometry (NR). The mAb tested (“mAb-1”) showed highest surface loading to silica at pH 7.4 (~12 mg/m2), with lower surface loading at pH 5.5 (~5.5 mg/m2, further from its pI of 8.99) and to hydrophobized silica (~2 mg/m2). The extent of desorption of mAb-1 from silica or hydrophobized silica was related to the relative affinity of polysorbate 20 or 80 for the same surface. mAb-1 adsorbed to silica on co-injection with polysorbate (above its critical micelle concentration) and also to silica pre-coated with polysorbate. A bilayer model was developed from NR data for mAb-1 at concentrations of 50–5000 mg/L, pH 5.5, and 50–2000 mg/L, pH 7.4. The inner mAb-1 layer was adsorbed to the SiO2 surface at near saturation with an end-on” orientation, while the outer mAb-1 layer was sparse and molecules had a “side-on” orientation. A non-uniform triple layer was observed at 5000 mg/L, pH 7.4, suggesting mAb-1 adsorbed to the SiO2 surface as oligomers at this concentration and pH. mAb-1 adsorbed as a sparse monolayer to hydrophobized silica, with a layer thickness increasing with bulk concentration - suggesting a near end-on orientation without observable relaxation-unfolding. PMID:23196810
Purification and characterisation of the fission yeast Ndc80 complex.
Matsuo, Yuzy; Maurer, Sebastian P; Surrey, Thomas; Toda, Takashi
2017-07-01
The Ndc80 complex is a conserved outer kinetochore protein complex consisting of Ndc80 (Hec1), Nuf2, Spc24 and Spc25. This complex comprises a major, if not the sole, platform with which the plus ends of the spindle microtubules directly interact. In fission yeast, several studies indicate that multiple microtubule-associated proteins including the Dis1/chTOG microtubule polymerase and the Mal3/EB1 microtubule plus-end tracking protein directly or indirectly bind Ndc80, thereby ensuring stable kinetochore-microtubule attachment. However, the purification of the Ndc80 complex from this yeast has not been achieved, which hampers the in-depth investigation as to how the outer kinetochore attaches to the plus end of the spindle microtubule. Here we report the two-step purification of the fission yeast Ndc80 holo complex from bacteria. First, we purified separately two sub-complexes consisting of Ndc80-Nuf2 and Spc24-Spc25. Then, these two sub-complexes were mixed and applied to size-exclusion chromatography. The reconstituted Ndc80 holo complex is composed of four subunits with equal stoichiometry. The complex possesses microtubule-binding activity, and Total Internal Reflection Fluorescence (TIRF)-microscopy assays show that the complex binds the microtubule lattice. Interestingly, unlike the human complex, the fission yeast complex does not track depolymerising microtubule ends. Further analysis shows that under physiological ionic conditions, the Ndc80 holo complex does not detectably bind Dis1, but instead it interacts with Mal3/EB1, by which the Ndc80 complex tracks the growing microtubule plus end. This result substantiates the notion that the Ndc80 complex plays a crucial role in establishment of the dynamic kinetochore-microtubule interface by cooperating with chTOG and EB1. Copyright © 2017 The Francis Crick Institute. Published by Elsevier Inc. All rights reserved.
NASA Astrophysics Data System (ADS)
Blecha, V.
A new Bouguer anomaly map of western part of southern Yemen margin has been compiled. Densities of rock samples from main geological units (Precambrian base- ment, Mesozoic sediments, Tertiary volcanites) have been measured and used for grav- ity modeling. Regional gravity map indicates decrease of thickness of continental crust from volcanites of the Yemen Trap Series towards the coast of the Gulf of Aden. Most remarkable feature in the map of residual anomalies is a positive anomaly over the Dhala graben. The Dhala graben is a prominent geological structure in the area of study trending parallel to the Red Sea axis. Gravity modeling on a profile across the Dhala graben presumes intrusive plutonic rocks beneath the graben. There are two other areas in the southwestern tip of Arabia, which have essentially the same struc- tural position as the Dhala graben: the Jabal Tirf volcanic rift zone in the southern Saudi Arabia and Jabal Hufash extensional zone in northern Yemen. All three areas extend along the line trending parallel to the Red Sea axis with length of about 500 km. The line coincides with the axis of Afar (Danakil) depression after Arabia is shifted and rotated back to Africa. These facts imply conclusion that the Oligocene - Early Miocene magmatic activity on the Jabal Tirf - Dhala lineament is related to the same original deep tectonic zone, forming present-day Afar depression and still active.
Trends in alcohol-impaired driving in Canada.
Vanlaar, Ward; Robertson, Robyn; Marcoux, Kyla; Mayhew, Daniel; Brown, Steve; Boase, Paul
2012-09-01
While a general decreasing trend in the number of persons killed in a traffic crash involving a drinking driver has occurred in Canada since the 1980s, it is evident that much of this decrease occurred in the 1990s. Since 2002, less progress has been made as the number of persons killed in crashes involving drinking drivers remains high. To better understand the current situation, this paper describes trends in drinking and driving in Canada from 1998 to 2011 using multiple indicators based on data collected for the Traffic Injury Research Foundation's (TIRF) Road Safety Monitor (RSM), the National Opinion Poll on Drinking and Driving, and trends in alcohol-related crashes based on data collected for TIRF's national Fatality Database in Canada. There has been a continued and consistent decrease in the number of fatalities involving a drinking driver in Canada. This remains true when looking at the number of fatalities involving a drinking driver per 100,000 population and per 100,000 licensed drivers. This decreasing trend is also still apparent when considering the percentage of persons killed in a traffic crash in Canada involving a drinking driver although less pronounced. Data from the RSM further show that the percentage of those who reported driving after they thought they were over the legal limit has also declined. However, regardless of the apparent decreasing trend in drinking driving fatalities and behaviour, reductions have been relatively modest, and fatalities in crashes involving drivers who have consumed alcohol remain high at unacceptable levels. Copyright © 2012 Elsevier Ltd. All rights reserved.
Src-dependent phosphorylation of caveolin-1 Tyr-14 promotes swelling and release of caveolae
Zimnicka, Adriana M.; Husain, Yawer S.; Shajahan, Ayesha N.; Sverdlov, Maria; Chaga, Oleg; Chen, Zhenlong; Toth, Peter T.; Klomp, Jennifer; Karginov, Andrei V.; Tiruppathi, Chinnaswamy; Malik, Asrar B.; Minshall, Richard D.
2016-01-01
Caveolin 1 (Cav1) is a required structural component of caveolae, and its phosphorylation by Src is associated with an increase in caveolae-mediated endocytosis. Here we demonstrate, using quantitative live-cell 4D, TIRF, and FRET imaging, that endocytosis and trafficking of caveolae are associated with a Cav1 Tyr-14 phosphorylation-dependent conformational change, which spatially separates, or loosens, Cav1 molecules within the oligomeric caveolar coat. When tracked by TIRF and spinning-disk microscopy, cells expressing phosphomimicking Cav1 (Y14D) mutant formed vesicles that were greater in number and volume than with Y14F-Cav1-GFP. Furthermore, we observed in HEK cells cotransfected with wild-type, Y14D, or Y14F Cav1-CFP and -YFP constructs that FRET efficiency was greater with Y14F pairs than with Y14D, indicating that pY14-Cav1 regulates the spatial organization of Cav1 molecules within the oligomer. In addition, albumin-induced Src activation or direct activation of Src using a rapamycin-inducible Src construct (RapR-Src) led to an increase in monomeric Cav1 in Western blots, as well as a simultaneous increase in vesicle number and decrease in FRET intensity, indicative of a Src-mediated conformational change in CFP/YFP-tagged WT-Cav1 pairs. We conclude that phosphorylation of Cav1 leads to separation or “spreading” of neighboring negatively charged N-terminal phosphotyrosine residues, promoting swelling of caveolae, followed by their release from the plasma membrane. PMID:27170175
Olshina, Maya A; Angrisano, Fiona; Marapana, Danushka S; Riglar, David T; Bane, Kartik; Wong, Wilson; Catimel, Bruno; Yin, Meng-Xin; Holmes, Andrew B; Frischknecht, Friedrich; Kovar, David R; Baum, Jake
2015-07-18
Gliding motility in Plasmodium parasites, the aetiological agents of malaria disease, is mediated by an actomyosin motor anchored in the outer pellicle of the motile cell. Effective motility is dependent on a parasite myosin motor and turnover of dynamic parasite actin filaments. To date, however, the basis for directional motility is not known. Whilst myosin is very likely orientated as a result of its anchorage within the parasite, how actin filaments are orientated to facilitate directional force generation remains unexplained. In addition, recent evidence has questioned the linkage between actin filaments and secreted surface antigens leaving the way by which motor force is transmitted to the extracellular milieu unknown. Malaria parasites possess a markedly reduced repertoire of actin regulators, among which few are predicted to interact with filamentous (F)-actin directly. One of these, PF3D7_1251200, shows strong homology to the coronin family of actin-filament binding proteins, herein referred to as PfCoronin. Here the N terminal beta propeller domain of PfCoronin (PfCor-N) was expressed to assess its ability to bind and bundle pre-formed actin filaments by sedimentation assay, total internal reflection fluorescence (TIRF) microscopy and confocal imaging as well as to explore its ability to bind phospholipids. In parallel a tagged PfCoronin line in Plasmodium falciparum was generated to determine the cellular localization of the protein during asexual parasite development and blood-stage merozoite invasion. A combination of biochemical approaches demonstrated that the N-terminal beta-propeller domain of PfCoronin is capable of binding F-actin and facilitating formation of parallel filament bundles. In parasites, PfCoronin is expressed late in the asexual lifecycle and localizes to the pellicle region of invasive merozoites before and during erythrocyte entry. PfCoronin also associates strongly with membranes within the cell, likely mediated by interactions with phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) at the plasma membrane. These data suggest PfCoronin may fulfil a key role as the critical determinant of actin filament organization in the Plasmodium cell. This raises the possibility that macro-molecular organization of actin mediates directional motility in gliding parasites.
Tsg101 regulates PI(4,5)P2/Ca2+ signaling for HIV-1 Gag assembly
Ehrlich, Lorna S.; Medina, Gisselle N.; Photiadis, Sara; Whittredge, Paul B.; Watanabe, Susan; Taraska, Justin W.; Carter, Carol A.
2014-01-01
Our previous studies identified the 1,4,5-inositol trisphosphate receptor (IP3R), a channel mediating release of Ca2+ from ER stores, as a cellular factor differentially associated with HIV-1 Gag that might facilitate ESCRT function in virus budding. Channel opening requires activation that is initiated by binding of 1,4,5-triphosphate (IP3), a product of phospholipase C (PLC)-mediated PI(4,5)P2 hydrolysis. The store emptying that follows stimulates store refilling which requires intact PI(4,5)P2. Raising cytosolic Ca2+ promotes viral particle production and our studies indicate that IP3R and the ER Ca2+ store are the physiological providers of Ca2+ for Gag assembly and release. Here, we show that Gag modulates ER store gating and refilling. Cells expressing Gag exhibited a higher cytosolic Ca2+ level originating from the ER store than control cells, suggesting that Gag induced release of store Ca2+. This property required the PTAP motif in Gag that recruits Tsg101, an ESCRT-1 component. Consistent with cytosolic Ca2+ elevation, Gag accumulation at the plasma membrane was found to require continuous IP3R activation. Like other IP3R channel modulators, Gag was detected in physical proximity to the ER and to endogenous IP3R, as indicated respectively by total internal reflection fluorescence (TIRF) and immunoelectron microscopy (IEM) or indirect immunofluorescence. Reciprocal co-immunoprecipitation suggested that Gag and IP3R proximity is favored when the PTAP motif in Gag is intact. Gag expression was also accompanied by increased PI(4,5)P2 accumulation at the plasma membrane, a condition favoring store refilling capacity. Supporting this notion, Gag particle production was impervious to treatment with 2-aminoethoxydiphenyl borate, an inhibitor of a refilling coupling interaction. In contrast, particle production by a Gag mutant lacking the PTAP motif was reduced. We conclude that a functional PTAP L domain, and by inference Tsg101 binding, confers Gag with an ability to modulate both ER store Ca2+ release and ER store refilling. PMID:24904548
Biomimetic oligosaccharide and peptide surfactant polymers designed for cardiovascular biomaterials
NASA Astrophysics Data System (ADS)
Ruegsegger, Mark Andrew
A common problem associated with cardiovascular devices is surface induced thrombosis initiated by the rapid, non-specific adsorption of plasma proteins onto the biomaterial surface. Control of the initial protein adsorption is crucial to achieve the desired longevity of the implanted biomaterial. The cell membrane glycocalyx acts as a non-thrombogenic interface through passive (dense oligosaccharide structures) and active (ligand/receptor interactions) mechanisms. This thesis is designed to investigate biomimicry of the cell glycocalyx to minimize non-specific protein adsorption and promote specific ligand/receptor interactions. Biomimetic macromolecules were designed through the molecular-scale engineering of polymer surfactants, utilizing a poly(vinyl amine) (PVAm) backbone to which hydrophilic (dextran, maltose, peptide) and hydrophobic alkyl (hexanoyl or hexanal) chains are simultaneously attached. The structure was controlled through the molar feed ratio of hydrophobic-to-hydrophilic groups, which also provided control of the solution and surface-active properties. To mimic passive properties, a series of oligomaltose surfactants were synthesized with increasing saccharide length (n = 2, 7, 15 where n is number of glucose units) to investigate the effect of coating height on protein adsorption. The surfactants were characterized by infra red (IR) and nuclear magnetic resonance (NMR) spectroscopies for structural properties and atomic force microscopy (AFM) and contact angle goniometry for surface activity. Protein adsorption under dynamic flow (5 dyn/cm2) was reduced by 85%--95% over the bare hydrophobic substrate; platelet adhesion dropped by ˜80% compared to glass. Peptide ligands were incorporated into the oligosaccharide surfactant to promote functional activity of the passive coating. The surfactants were synthesized to contain 0%, 25%, 50%, 75%, and 100% peptide ligand density and were stable on hydrophobic surfaces. The peptide surface density was calculated to be 0.86 ligands/nm2 for PVAm(Pep)(100%), as determined by total internal reflection fluorescence (TIRF) spectroscopy. Similar cell growth was observed on the 100% peptide surfactant as for the fibronectin control, and no cell growth was seen on the 0% peptide. Increasing cell viability was observed for the surfaces with increasing peptide density. These results indicate much promise for surfactant polymers in surface modification and the capability to mimic the passive and active properties of the cell glycocalyx.
Reconnaissance geologic map of the Wadi Khulab Quadrangle, sheet 16/43 A, Kingdom of Saudi Arabia
Blank, Horace Richard; Gettings, Mark E.
1985-01-01
From west to east, the physiography of the mapped area consists of a portion of the Tiharmat Asir, or coastal plain, extending from the Jizan quadrangle to the west (Blank and Gettings, 1984), a northwest-trending hill range close to the western border of the quadrangle; a pediplain; and the foothills of the Red Sea escarpment. The top of the escarpment is about 50 km east of the mapped area, in the Yemen Arab Republic. Within Saudi Arabia, the highest elevation in the quadrangle about 77 m above sea level, is found at Tirf in the western range of hills.
Advanced Methods in Fluorescence Microscopy
Fritzky, Luke; Lagunoff, David
2013-01-01
It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres. PMID:23271142
Advanced methods in fluorescence microscopy.
Fritzky, Luke; Lagunoff, David
2013-01-01
It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbe limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.
Advanced methods in fluorescence microscopy.
Fritzky, Luke; Lagunoff, David
2013-01-01
It requires a good deal of will power to resist hyperbole in considering the advances that have been achieved in fluorescence microscopy in the last 25 years. Our effort has been to survey the modalities of microscopic fluorescence imaging available to cell biologists and perhaps useful for diagnostic pathologists. The gamut extends from established confocal laser scanning through multiphoton and TIRF to the emerging technologies of super-resolution microscopy that breech the Abbé limit of resolution. Also considered are the recent innovations in structured and light sheet illumination, the use of FRET and molecular beacons that exploit specific characteristics of designer fluorescent proteins, fluorescence speckles, and second harmonic generation for native anisometric structures like collagen, microtubules and sarcomeres.
Flagellar generated flow mediates attachment of Giardia lamblia
NASA Astrophysics Data System (ADS)
Urbach, Jeffrey; Luo, Haibei; Picou, Theodore; McAllister, Ryan; Elmendorf, Heidi
2011-03-01
Giardia lamblia is a protozoan parasite responsible for widespread diarrheal disease in humans and animals worldwide. Attachment to the host intestinal mucosa and resistance to peristalsis is necessary for establishing infection, but the physical basis for this attachment is poorly understood. We report results from TIRF and confocal fluorescence microscopy that demonstrate that the regular beating of the posterior flagella generate a flow through the ventral disk, a suction-cup shaped structure that is against the substrate during attachment. Finite element simulations are used to compare the negative pressure generated by the flow to the measured attachment force and the expected performance of the flagellar pump. NIH grant 1R21AI062934-0.
Frustrated Total Internal Reflection: A Simple Application and Demonstration.
ERIC Educational Resources Information Center
Zanella, F. P.; Magalhaes, D. V.; Oliveira, M. M.; Bianchi, R. F.; Misoguti, L.; Mendonca, C. R.
2003-01-01
Describes the total internal reflection process that occurs when the internal angle of incidence is equal to or greater than the critical angle. Presents a demonstration of the effect of frustrated total internal reflection (FTIR). (YDS)
Structure and activity of full-length formin mDia1
Maiti, Sankar; Michelot, Alphee; Gould, Christopher; Blanchoin, Laurent; Sokolova, Olga; Goode, Bruce L.
2012-01-01
Formins are a conserved family of actin assembly-promoting factors with essential and diverse biological roles. Most of our biochemical understanding of formin effects on actin dynamics is derived from studies using formin fragments. In addition, all structural information on formins has been limited to fragments. This has left open key questions about the structure, activity and regulation of intact formin proteins. Here, we isolated full-length mouse mDia1 (mDia1-FL) and found that it forms tightly autoinhibited dimers that can only be partially activated by RhoA. We solved the structure of autoinhibited mDia1-FL using electron microscopy and single particle analysis. Docking of crystal structures into the 3D reconstruction revealed that the fork-shaped N-terminal DID-CC region hangs over the ring-shaped FH2 domain, suggesting that autoinhibition results from steric obstruction of actin binding. Deletion of the C-terminal DAD domain extended mDia1 structure and activated it for actin assembly. Using TIRF microscopy, we observed that RhoA-activated mDia1-FL persistently accelerated filament elongation in the presence of profilin similar to mDia1 FH1-FH2 fragment. These observations validate the known activities of FH1-FH2 fragments as reflecting those of the intact molecule. Our results further suggest that mDia1-FL does not readily snap back into the autoinhibited conformation and dissociate from growing filament ends, and thus additional factors may be required to displace formins and restrict filament length. PMID:22605659
Prisms with total internal reflection as solar reflectors
Rabl, Arnulf; Rabl, Veronika
1978-01-01
An improved reflective wall for radiant energy collection and concentration devices is provided. The wall is comprised of a plurality of prisms whose frontal faces are adjacent and which reflect the desired radiation by total internal reflection.
Wang, Shinn-Fwu; Chiu, Ming-Hung; Chen, Wei-Wu; Kao, Fu-Hsi; Chang, Rong-Seng
2009-05-01
A small-displacement sensing system based on multiple total internal reflections in heterodyne interferometry is proposed. In this paper, a small displacement can be obtained only by measuring the variation in phase difference between s- and p-polarization states for the total internal reflection effect. In order to improve the sensitivity, we increase the number of total internal reflections by using a parallelogram prism. The theoretical resolution of the method is better than 0.417 nm. The method has some merits, e.g., high resolution, high sensitivity, and real-time measurement. Also, its feasibility is demonstrated.
Using Video Records to Mediate Teaching Interns' Critical Reflection
ERIC Educational Resources Information Center
Scott, Sarah E.; Kucan, Linda; Correnti, Richard; Miller, Leigh A.
2013-01-01
In this study we investigated how the use of video records in a literacy methods course supports the development of reflective practitioners when video is a core element of the course design. Specifically, we detail how interns' video-based reflections provide evidence that the use of video records of teaching interns' promotes the development of…
NASA Astrophysics Data System (ADS)
Lopez, Benjamin
2015-03-01
EB1 is an important microtubule associating protein (MAP) that acts as a master coordinator of protein activity at the growing plus-end of the microtubule. We can recapitulate the plus-end binding behavior of EB1 along the entire length of a static microtubule using microtubules polymerized in the presence of the nonhydrolyzable GTP analogs GMPCPP and GTP γS instead of GTP. Through the use of single-molecule TIRF imaging we find that EB1 is highly dynamic (with a sub-second characteristic binding lifetime) and continuously diffusive while bound to the microtubule. We measure the diffusion coefficient, D, through linear fitting to mean-squared displacement of individually labeled proteins, and the binding lifetime, τ, by fitting a single exponential decay to the probability distribution of trajectory lifetimes. In agreement with measurements of other diffusive MAPs, we find that D increases and τ decreases with increasing ionic strength. We also find that D is sensitive to the choice of GTP analog: EB1 proteins bound to GTP γS polymerized microtubules have a D half of that found with GMPCPP polymerized microtubules. To compare these single-molecule measurements to the bulk binding behavior of EB1, we use TIRF imaging to measure the intensity of microtubules coated with EB1-GFP as a function of EB1 concentration. We find that EB1 binding is cooperative and both the quantity of EB1 bound and the dissociation constant are sensitive to GTP analog and ionic concentration. The correlation between binding affinity and D and the cooperative nature of EB1-microtubule binding leads to a decrease in D with increasing EB1 concentration. Interestingly, we also find an increase in τ at high EB1 concentrations, consistent with attractive EB1-microtubule interactions driving the cooperativity. To further understand the nature of the cooperativity we estimate the interaction energy by measuring the association and dissociation rates (kon and koff respectively) at different concentrations of EB1.
Prevalence and trends of drugged driving in Canada.
Robertson, Robyn D; Mainegra Hing, Marisela; Pashley, Charlotte R; Brown, Steve W; Vanlaar, Ward G M
2017-02-01
This study evaluates prevalence and trends in drugged driving in Canada based on multiple indicators collected from the Road Safety Monitor (RSM) and Canada's National Fatality Database maintained by the Traffic Injury Research Foundation (TIRF). The objective of this paper is to identify the state of drug-positive driving in Canada, as well as to make comparisons with data from previous years to determine whether changes have occurred. Available data from the RSM on self-reported drugged driving behaviours were collected and analyzed using multivariate techniques in various years spanning from 2002 to 2015. Data from TIRF's National Fatality Database from 2000 to 2012 were also analyzed to evaluate trends and prevalence of drugs in fatally injured drivers across Canada. Additionally, differences among drugged drivers with respect to gender and age were studied. Analyses of the RSM data and of the National Fatality Database showed that, as a whole, the prevalence of drugged driving has remained relatively stable over the past decade, with some changes noticed in specific years for some drug types. Specifically from the RSM, there was a 62.5% increase from the 1.6% of drivers reporting driving within two hours of using marijuana in 2013 to 2.6% in 2015. The analyses of the fatality data revealed a 16.9% increase in the percentage of fatally injured drivers testing positive for drugs between 2000 and 2012 (from 33.56% to 39.24%). Cocaine-positive fatally injured drivers increased from 3.6% in 2000 to 6.2% in 2012. Similarly, marijuana-positive fatally injured drivers increased from 12.8% in 2000 to 19.7% in 2012. Results showed varying characteristics with respect to gender and age among self-reported and fatally injured drugged drivers. Drugged driving behaviours remain prevalent among Canadian drivers and drugs continue to be found in over one-third of tested fatally injured drivers. Although self-reported behaviours have neither decreased nor increased overall in the past decade according to RSM data, with the exception of driving within two hours of using marijuana, data from fatally injured drivers reveal that small, but significant increases in some behaviours have occurred. Copyright © 2016 Elsevier Ltd. All rights reserved.
An Arduino-Based Experiment Designed to Clarify the Transition to Total Internal Reflection
ERIC Educational Resources Information Center
Atkin, Keith
2018-01-01
The topic of refraction and reflection of light at the boundary of transparent media is a fundamentally important one. The special case of total internal reflection is however commonly misrepresented in elementary textbooks. This paper addresses the problem and describes an experimental procedure for measuring and displaying reflected and…
Studies of the Reflection, Refraction and Internal Reflection of Light
ERIC Educational Resources Information Center
Lanchester, P. C.
2014-01-01
An inexpensive apparatus and associated experiments are described for studying the basic laws of reflection and refraction of light at an air-glass interface, and multiple internal reflections within a glass block. In order to motivate students and encourage their active participation, a novel technique is described for determining the refractive…
Photovoltaic module with light reflecting backskin
Gonsiorawski, Ronald C [Danvers, MA
2007-07-03
A photovoltaic module comprises electrically interconnected and mutually spaced photovoltaic cells that are encapsulated by a light-transmitting encapsulant between a light-transparent front cover and a back cover, with the back cover sheet being an ionomer/nylon alloy embossed with V-shaped grooves running in at least two directions and coated with a light reflecting medium so as to provide light-reflecting facets that are aligned with the spaces between adjacent cells and oriented so as to reflect light falling in those spaces back toward said transparent front cover for further internal reflection onto the solar cells, whereby substantially all of the reflected light will be internally reflected from said cover sheet back to the photovoltaic cells, thereby increasing the current output of the module. The internal reflector improves power output by as much as 67%.
ERIC Educational Resources Information Center
Le Cornu, Alison
2009-01-01
The study of the process of reflection has a dignified history. However, few have linked reflection to the development of the self in such a way that the form of reflection is understood to influence the resultant type of self. This article explores the process of reflection using a framework of meaning making, internalization, and externalization…
ERIC Educational Resources Information Center
Furman, Rich; Coyne, Ann; Negi, Nalini Junko
2008-01-01
This descriptive article explores the uses of poetry and journaling exercises as means of helping students develop their self-reflective capacities within the context of international social work. First, self-reflection and its importance to social work practice and education is discussed. Second, the importance of self-reflection in international…
Developing DNA nanotechnology using single-molecule fluorescence.
Tsukanov, Roman; Tomov, Toma E; Liber, Miran; Berger, Yaron; Nir, Eyal
2014-06-17
CONSPECTUS: An important effort in the DNA nanotechnology field is focused on the rational design and manufacture of molecular structures and dynamic devices made of DNA. As is the case for other technologies that deal with manipulation of matter, rational development requires high quality and informative feedback on the building blocks and final products. For DNA nanotechnology such feedback is typically provided by gel electrophoresis, atomic force microscopy (AFM), and transmission electron microscopy (TEM). These analytical tools provide excellent structural information; however, usually they do not provide high-resolution dynamic information. For the development of DNA-made dynamic devices such as machines, motors, robots, and computers this constitutes a major problem. Bulk-fluorescence techniques are capable of providing dynamic information, but because only ensemble averaged information is obtained, the technique may not adequately describe the dynamics in the context of complex DNA devices. The single-molecule fluorescence (SMF) technique offers a unique combination of capabilities that make it an excellent tool for guiding the development of DNA-made devices. The technique has been increasingly used in DNA nanotechnology, especially for the analysis of structure, dynamics, integrity, and operation of DNA-made devices; however, its capabilities are not yet sufficiently familiar to the community. The purpose of this Account is to demonstrate how different SMF tools can be utilized for the development of DNA devices and for structural dynamic investigation of biomolecules in general and DNA molecules in particular. Single-molecule diffusion-based Förster resonance energy transfer and alternating laser excitation (sm-FRET/ALEX) and immobilization-based total internal reflection fluorescence (TIRF) techniques are briefly described and demonstrated. To illustrate the many applications of SMF to DNA nanotechnology, examples of SMF studies of DNA hairpins and Holliday junctions and of the interactions of DNA strands with DNA origami and origami-related devices such as a DNA bipedal motor are provided. These examples demonstrate how SMF can be utilized for measurement of distances and conformational distributions and equilibrium and nonequilibrium kinetics, to monitor structural integrity and operation of DNA devices, and for isolation and investigation of minor subpopulations including malfunctioning and nonreactive devices. Utilization of a flow-cell to achieve measurements of dynamics with increased time resolution and for convenient and efficient operation of DNA devices is discussed briefly. We conclude by summarizing the various benefits provided by SMF for the development of DNA nanotechnology and suggest that the method can significantly assist in the design and manufacture and evaluation of operation of DNA devices.
Harnessing Multiple Internal Reflections to Design Highly Absorptive Acoustic Metasurfaces
NASA Astrophysics Data System (ADS)
Shen, Chen; Cummer, Steven A.
2018-05-01
The rapid development of metasurfaces has enabled numerous intriguing applications with acoustically thin sheets. Here we report the theory and experimental realization of a nonresonant sound-absorbing strategy using metasurfaces by harnessing multiple internal reflections. We theoretically and numerically show that the higher-order diffraction of thin gradient-index metasurfaces is tied to multiple internal reflections inside the unit cells. Highly absorbing acoustic metasurfaces can be realized by enforcing multiple internal reflections together with a small amount of loss. A reflective gradient-index acoustic metasurface is designed based on the theory, and we further experimentally verify the performance using a three-dimensional printed prototype. Measurements show over 99% energy absorption at the peak frequency and a 95% energy absorption bandwidth of around 600 Hz. The proposed mechanism provides an alternative route for sound absorption without the necessity of high absorption of the individual unit cells.
Internal high-reflectivity omni-directional reflectors
NASA Astrophysics Data System (ADS)
Xi, J.-Q.; Ojha, Manas; Plawsky, J. L.; Gill, W. N.; Kim, Jong Kyu; Schubert, E. F.
2005-07-01
An internal high-reflectivity omni-directional reflector (ODR) for the visible spectrum is realized by the combination of total internal reflection using a low-refractive-index (low-n) material and reflection from a one-dimensional photonic crystal (1D PC). The low-n layer limits the range of angles in the 1D PC to values below the Brewster angle, thereby enabling high reflectivity and omni-directionality. This ODR is demonstrated using GaP as ambient, nanoporous SiO2 with a very low refractive index (n=1.10), and a four-pair TiO2/SiO2 multilayer stack. The results indicate a two orders of magnitude lower angle-integrated transverse-electric-transverse-magnetic polarization averaged mirror loss of the ODR compared with conventional distributed Bragg reflectors and metal reflectors. This indicates the high potential of the internal ODRs for optoelectronic semiconductor devices, e.g., light-emitting diodes.
Passively Q-switched side pumped monolithic ring laser
NASA Technical Reports Server (NTRS)
Li, Steven X. (Inventor)
2012-01-01
Disclosed herein are systems and methods for generating a side-pumped passively Q-switched non-planar ring oscillator. The method introduces a laser into a cavity of a crystal, the cavity having a round-trip path formed by a reflection at a dielectrically coated front surface, a first internal reflection at a first side surface of the crystal at a non-orthogonal angle with the front, a second internal reflection at a top surface of the crystal, and a third internal reflection at a second side surface of the crystal at a non-orthogonal angle with the front. The method side pumps the laser at the top or bottom surface with a side pump diode array beam and generates an output laser emanating at a location on the front surface. The design can include additional internal reflections to increase interaction with the side pump. Waste heat may be removed by mounting the crystal to a heatsink.
Seismic reflection imaging, accounting for primary and multiple reflections
NASA Astrophysics Data System (ADS)
Wapenaar, Kees; van der Neut, Joost; Thorbecke, Jan; Broggini, Filippo; Slob, Evert; Snieder, Roel
2015-04-01
Imaging of seismic reflection data is usually based on the assumption that the seismic response consists of primary reflections only. Multiple reflections, i.e. waves that have reflected more than once, are treated as primaries and are imaged at wrong positions. There are two classes of multiple reflections, which we will call surface-related multiples and internal multiples. Surface-related multiples are those multiples that contain at least one reflection at the earth's surface, whereas internal multiples consist of waves that have reflected only at subsurface interfaces. Surface-related multiples are the strongest, but also relatively easy to deal with because the reflecting boundary (the earth's surface) is known. Internal multiples constitute a much more difficult problem for seismic imaging, because the positions and properties of the reflecting interfaces are not known. We are developing reflection imaging methodology which deals with internal multiples. Starting with the Marchenko equation for 1D inverse scattering problems, we derived 3D Marchenko-type equations, which relate reflection data at the surface to Green's functions between virtual sources anywhere in the subsurface and receivers at the surface. Based on these equations, we derived an iterative scheme by which these Green's functions can be retrieved from the reflection data at the surface. This iterative scheme requires an estimate of the direct wave of the Green's functions in a background medium. Note that this is precisely the same information that is also required by standard reflection imaging schemes. However, unlike in standard imaging, our iterative Marchenko scheme retrieves the multiple reflections of the Green's functions from the reflection data at the surface. For this, no knowledge of the positions and properties of the reflecting interfaces is required. Once the full Green's functions are retrieved, reflection imaging can be carried out by which the primaries and multiples are mapped to their correct positions, with correct reflection amplitudes. In the presentation we will illustrate this new methodology with numerical examples and discuss its potential and limitations.
An Arduino-based experiment designed to clarify the transition to total internal reflection
NASA Astrophysics Data System (ADS)
Atkin, Keith
2018-03-01
The topic of refraction and reflection of light at the boundary of transparent media is a fundamentally important one. The special case of total internal reflection is however commonly misrepresented in elementary textbooks. This paper addresses the problem and describes an experimental procedure for measuring and displaying reflected and transmitted light intensities using readily available components and the Arduino microcontroller.
New mounting improves solar-cell efficiency
NASA Technical Reports Server (NTRS)
Shepard, N. F., Jr.
1980-01-01
Method boosts output by about 20 percent by trapping and redirecting solar radiation without increasing module depth. Mounted solar-cell array is covered with internally reflecting plate. Plate is attached to each cell by transparent adhesive, and space between cells is covered with layer of diffusely reflecting material. Solar energy falling on space between cells is diffused and reflected internally by plate until it is reflected onto solar cell.
Nonimaging light concentration using total internal reflection films.
Ouellette, G; Waltham, C E; Drees, R M; Poon, A; Schubank, R; Whitehead, L A
1992-05-01
We present a method of fabricating nonimaging light concentrators from total internal reflection film. A prototype has been made and tested and found to operate in agreement with predictions of ray-tracing codes. The performance of the prototype is comparable with that of concentrators made from specular reflecting materials.
Georgopoulos, Michael; Zehetmayer, Martin; Ruhswurm, Irene; Toma-Bstaendig, Sabine; Ségur-Eltz, Nikolaus; Sacu, Stefan; Menapace, Rupert
2003-01-01
This study assesses differences in relative tumour regression and internal acoustic reflectivity after 3 methods of radiotherapy for uveal melanoma: (1) brachytherapy with ruthenium-106 radioactive plaques (RU), (2) fractionated high-dose gamma knife stereotactic irradiation in 2-3 fractions (GK) or (3) fractionated linear-accelerator-based stereotactic teletherapy in 5 fractions (Linac). Ultrasound measurements of tumour thickness and internal reflectivity were performed with standardised A scan pre-operatively and 3, 6, 9, 12, 18, 24 and 36 months postoperatively. Of 211 patients included in the study, 111 had a complete 3-year follow-up (RU: 41, GK: 37, Linac: 33). Differences in tumour thickness and internal reflectivity were assessed with analysis of variance, and post hoc multiple comparisons were calculated with Tukey's honestly significant difference test. Local tumour control was excellent with all 3 methods (>93%). At 36 months, relative tumour height reduction was 69, 50 and 30% after RU, GK and Linac, respectively. In all 3 treatment groups, internal reflectivity increased from about 30% initially to 60-70% 3 years after treatment. Brachytherapy with ruthenium-106 plaques results in a faster tumour regression as compared to teletherapy with gamma knife or Linac. Internal reflectivity increases comparably in all 3 groups. Besides tumour growth arrest, increasing internal reflectivity is considered as an important factor indicating successful treatment. Copyright 2003 S. Karger AG, Basel
Character of the opposition effect and negative polarization
NASA Technical Reports Server (NTRS)
Pieters, Carle M.; Shkuratov, Yu. G.; Stankevich, D. G.
1991-01-01
Photometric and polarimetric properties at small phase angles were measured for silicates with controlled surface properties in order to distinguish properties that are associated with surface reflection from those that are associated with multiple scattering from internal grain boundaries. These data provide insight into the causes and conditions of photometric properties observed at small phase angles for dark bodies of the solar system. Obsidian was chosen to represent a silicate dielectric with no internal scattering boundaries. Because obsidian is free of internal scatterers, light reflected from both the rough and smooth obsidian samples is almost entirely single and multiple Fresnel reflections form surface facets with no body component. Surface structure alone cannot produce an opposition effect. Comparison of the obsidian and basalt results indicates that for an opposition effect to occur, surface texture must be both rough and contain internal scattering interfaces. Although the negative polarization observed for the obsidian samples indicates single and multiple reflections are part of negative polarization, the longer inversion angle of the multigrain inversion samples implies that internal reflections must also contribute a significant negative polarization component.
Germania Quo Vadis?: Dynamics of Change in German Security Policy
2007-06-01
Keohane, Robert O., Michael Brecher and Frank Harveys, eds. Institutional Theory in international Relations. In: Millennial Reflections on International...Hegemony: Cooperation and Discord in the World Political Economy, (Princeton University Press, 1984), pp. 78-109; Robert O. Keohane, “ Institutional ... Theory in international Relations,” in Michael Brecher and Frank Harveys, eds., Millennial Reflections on International Studies (University of Michigan
Joyce-McCoach, Joanne T; Parrish, Dominique R; Andersen, Patrea R; Wall, Natalie
2013-09-01
Being reflective is well established as an important conduit of practice development, a desirable tertiary graduate quality and a core competency of health professional membership. By assisting students to be more effective in their ability to reflect, they are better able to formulate strategies to manage issues experienced within a professional context, which ultimately assists them to be better service providers. However, some students are challenged by the practice of reflection and these challenges are even more notable for international students. This paper presents a teaching initiative that focused specifically on enhancing the capacity of an international cohort of nursing students, to engage in reflective practice. The initiative centered on an evaluation of a reflective practice core subject, which was taught in a Master of Nursing programme delivered in Hong Kong. A learning-centered framework was used to evaluate the subject and identify innovative strategies that would better assist international students to develop reflective practices. The outcomes of curriculum and teaching analysis and proposed changes and innovations in teaching practice to support international students are presented and discussed. Copyright © 2012 Elsevier Ltd. All rights reserved.
Single Molecule Stepping and Structural Dynamics of Myosin X
Sun, Yujie; Sato, Osamu; Ruhnow, Felix; Arsenault, Mark E.; Ikebe, Mitsuo; Goldman, Yale E.
2010-01-01
Myosin X is an unconventional myosin with puzzling motility properties. We studied the motility of dimerized myosin X using single molecule fluorescence techniques – polTIRF, FIONA, and Parallax to measure rotation angles and 3-dimensional position of the molecule during its walk. It was found that Myosin X steps processively in a hand-over-hand manner following a left-handed helical path along both single actin filaments and bundles. Its step size and velocity are smaller on actin bundles than individual filaments, suggesting myosin X often steps onto neighboring filaments in a bundle. The data suggest that a previously postulated single α-helical domain mechanically extends the 3-IQ motif lever arm and either the neck-tail hinge or the tail is flexible. These structural features, in conjunction with the membrane and microtubule binding domains, enable myosin X to perform multiple functions on varied actin structures in cells. PMID:20364131
A self-assembling lanthanide molecular nanoparticle for optical imaging†
Brown, Katherine A.; Yang, Xiaoping; Schipper, Desmond; Hall, Justin W.; DePue, Lauren J.; Gnanam, Annie J.; Arambula, Jonathan F.; Jones, Jessica N.; Swaminathan, Jagannath; Dieye, Yakhya; Vadivelu, Jamuna; Chandler, Don J.; Marcotte, Edward M.; Sessler, Jonathan L.; Ehrlich, Lauren I. R.; Jones, Richard A.
2015-01-01
Chromophores that incorporate f-block elements have considerable potential for use in bioimaging applications because of their advantageous photophysical properties compared to organic dye, which are currently widely used. We are developing new classes of lanthanide-based self-assembling molecular nanoparticles as reporters for imaging and as multi-functional nanoprobes or nanosensors for use with biological samples. One class of these materials, which we call lanthanide “nano-drums”, are homogeneous 4d–4f clusters approximately 25 to 30 Å in diameter. These are capable of emitting from the visible to near-infrared wavelengths. Here, we present the synthesis, crystal structure, photophysical properties and comparative cytotoxicity data for a 32 metal Eu-Cd nano-drum [Eu8Cd24L12(OAc)48] (1). We also explored the imaging capabilities of this nano-drum using epifluorescence, TIRF, and two-photon microscopy platforms. PMID:25512085
NASA Astrophysics Data System (ADS)
Aksu, Anil A.
2017-09-01
In this paper, we have considered the non-linear effects arising due to the collision of incident and reflected internal wave beams. It has already been shown analytically [Tabaei et al., "Nonlinear effects in reflecting and colliding internal wave beams," J. Fluid Mech. 526, 217-243 (2005)] and numerically [Rodenborn et al., "Harmonic generation by reflecting internal waves," Phys. Fluids 23, 026601 (2011)] that the internal wave beam collision generates the higher harmonics and mean flow in a linear stratification. In this paper, similar to previous analytical work, small amplitude wave theory is employed; however, it is formulated from energetics perspective which allows considering internal wave beams as the product of slowly varying amplitude and fast complex exponential. As a result, the mean energy propagation equation for the second harmonic wave is obtained. Finally, a similar dependence on the angle of incidence is obtained for the non-linear energy transfer to the second harmonic with previous analyses. A possible physical mechanism for this angle dependence on the second harmonic generation is also discussed here. In addition to previous studies, the viscous effects are also included in the mean energy propagation equation for the incident, the reflecting, and the second harmonic waves. Moreover, even though the mean flow obtained here is only confined to the interaction region, it is also affected by viscosity via the decay in the incident and the reflecting internal wave beams. Furthermore, a framework for the non-linear harmonic generation in non-linear stratification is also proposed here.
Improved power efficiency for very-high-temperature solar-thermal-cavity receivers
McDougal, A.R.; Hale, R.R.
1982-04-14
This invention is an improved solar energy cavity receiver for exposing materials and components to high temperatures. The receiver includes a housing having an internal reflective surface defining a cavity and having an inlet for admitting solar radiation thereto. A photothermal absorber is positiond in the cavity to receive radiation from the inlet. A reflective baffle is positioned between the absorber and the inlet to severely restrict the re-radiation of energy through the inlet. The front surface of the baffle defines a narrow annulus with the internal reflective surface of the housing. The front surface of the baffle is contoured to reflect incoming radiation onto the internal surface of the housing, from which it is reflected through the annulus and onto the front surface of the absorber. The back surface of the baffle intercepts radiation from the front of the absorber. With this arrangement, a high percentage of the solar power input is retained in the cavity; thus, high internal temperatues are attained.
Power efficiency for very high temperature solar thermal cavity receivers
McDougal, Allan R.; Hale, Robert R.
1984-01-01
This invention is an improved solar energy cavity receiver for exposing materials and components to high temperatures. The receiver includes a housing having an internal reflective surface defining a cavity and having an inlet for admitting solar radiation thereto. A photothermal absorber is positioned in the cavity to receive radiation from the inlet. A reflective baffle is positioned between the absorber and the inlet to severely restrict the re-radiation of energy through the inlet. The front surface of the baffle defines a narrow annulus with the internal reflective surface of the housing. The front surface of the baffle is contoured to reflect incoming radiation onto the internal surface of the housing, from which it is reflected through the annulus and onto the front surface of the absorber. The back surface of the baffle intercepts infrared radiation from the front of the absorber. With this arrangement, a high percentage of the solar power input is retained in the cavity; thus, high internal temperatures are attained.
ERIC Educational Resources Information Center
Levine, Rachel B.; Kern, David E.; Wright, Scott M.
2008-01-01
Narrative writing has been used to promote reflection and increased self-awareness among physicians. The purpose of this study was to determine the impact of prompted narrative writing on reflection. Thirty-two interns at 9 internal medicine residency programs participated in a year-long qualitative study about personal growth beginning in July of…
International Mindedness through the Looking Glass: Reflections on a Concept
ERIC Educational Resources Information Center
Castro, Paloma; Lundgren, Ulla; Woodin, Jane
2015-01-01
The aim of this article is to report and reflect on a research project involving the conceptualization of the term "International Mindedness", which is used across a range of International Baccalaureate (IB) global and local contexts. The research process involved both a critical analysis of IB official documents and a literature review…
Zheng, Qinsi; Jockusch, Steffen; Zhou, Zhou; Altman, Roger B.; Zhao, Hong; Asher, Wesley; Holsey, Michael; Mathiasen, Signe; Geggier, Peter; Javitch, Jonathan A.
2017-01-01
Bright, long-lasting organic fluorophores enable a broad range of imaging applications. “Self-healing” fluorophores, in which intra-molecularly linked protective agents quench photo-induced reactive species, exhibit both enhanced photostability and biological compatibility. However, the self-healing strategy has yet to achieve its predicted potential, particularly in the presence of ambient oxygen where live-cell imaging studies must often be performed. To identify key bottlenecks in this technology that can be used to guide further engineering developments, we synthesized a series of Cy5 derivatives linked to the protective agent cyclooctatetraene (COT) and examined the photophysical mechanisms curtailing their performance. The data obtained reveal that the photostability of self-healing fluorophores is limited by reactivity of the COT protective agent. The addition of electron withdrawing substituents to COT reduced its susceptibility to reactions with molecular oxygen and the fluorophore to which it is attached and increased its capacity to participate in triplet energy transfer. Exploiting these insights, we designed and synthesized a suite of modified COT-fluorophores spanning the visible spectrum that exhibited markedly increased intra-molecular photostabilization. Under ambient oxygen conditions, the photostability of Cy3 and Cy5 fluorophore derivatives increased by 3- and 9-fold in vitro and by 2- and 6-fold in living cells, respectively. We further show that this approach can improve a silicon rhodamine fluorophore. These findings offer a clear strategy for achieving the full potential of the self-healing approach and its application to the gamut of fluorophore species commonly used for biomedical imaging. PMID:28377799
Real time infrared aerosol analyzer
Johnson, Stanley A.; Reedy, Gerald T.; Kumar, Romesh
1990-01-01
Apparatus for analyzing aerosols in essentially real time includes a virtual impactor which separates coarse particles from fine and ultrafine particles in an aerosol sample. The coarse and ultrafine particles are captured in PTFE filters, and the fine particles impact onto an internal light reflection element. The composition and quantity of the particles on the PTFE filter and on the internal reflection element are measured by alternately passing infrared light through the filter and the internal light reflection element, and analyzing the light through infrared spectrophotometry to identify the particles in the sample.
ERIC Educational Resources Information Center
Charters, Alexander N.
2012-01-01
This article presents the author's reflections on involvement with six UNESCO international conferences on adult education. As adult educators continue to look forward with enthusiasm to the future of adult and continuing education in an increasingly international society, the author argues that they need to continually remember that the mission…
University Employment Transitions in International Performing Arts: The Intern's Story
ERIC Educational Resources Information Center
Carneiro, Maria
2013-01-01
This article reflects my journey as a performing arts student and intern both in Portugal and abroad. It is not intended as a personal journal, but rather a reflection and an aftermath comment on my experiences and learning processes. First it provides a context regarding my university education in a Southern European country, against a previous…
Technology Experiences of Student Interns in a One to One Mobile Program
ERIC Educational Resources Information Center
Cullen, Theresa A.; Karademir, Tugra
2018-01-01
This article describes how a group of student intern teachers (n = 51) in a one to one teacher education iPad program were asked to reflect using Experience Sampling Method (ESM) on their use of technology in the classroom during internship. Interns also completed summative reflections and class discussions. Data collected both in online and…
Value of Reflected Light Microscopy in Teaching.
ERIC Educational Resources Information Center
Pasteris, Jill Dill
1983-01-01
Briefly reviews some optical and other physical properties of minerals that can be determined in reflected/incident light. Topics include optical properties of minerals, reflectance, internal reflections, color, bireflectance and reflection pleochroism, anisotropism, zonation, and reflected light microscopy as a teaching tool in undergraduate…
DIRC Dreams: Research Directions for the Next Generation of Internally Reflected Imaging Counters
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ratcliff, Blair N.
1999-08-17
Some conceptual design features of the total internally reflecting,imaging Cherenkov counter (DIRC) are described. Limits of the DIRC approach to particle identification, and a few features of alternative DIRC designs, are briefly explored.
Novel method in solving non-polarizing condition in frustrated total internal reflection layers
NASA Astrophysics Data System (ADS)
Shi, Jin Hui; Wang, Zheng Ping
2008-03-01
When used at oblique angles of incidence, the reflectance and transmittance of thin films exhibit strong polarization effects, particularly for the films inside a glass cube. However, the polarization effects are undesirable in many applications. Novel non-polarizing beam splitter designs are shown, non-polarizing beam splitters with unique optical thin films are achieved through combination of interference and frustrated total internal reflection, the non-polarizing condition expressions based on frustrated total internal reflection is derived, and applied examples of the non-polarizing beam splitters are also presented with the optimization technique and the results of Rp=(50+/-0.4)% and Rs=(50+/-0.4)% in the wavelength range of 500-600nm are obtained.
Frustrated total internal reflection acoustic field sensor
Kallman, Jeffrey S.
2000-01-01
A frustrated total internal reflection acoustic field sensor which allows the acquisition of the acoustic field over an entire plane, all at once. The sensor finds use in acoustic holography and acoustic diffraction tomography. For example, the sensor may be produced by a transparent plate with transparent support members tall enough to support one or more flexible membranes at an appropriate height for frustrated total internal reflection to occur. An acoustic wave causes the membrane to deflect away from its quiescent position and thus changes the amount of light that tunnels through the gap formed by the support members and into the membrane, and so changes the amount of light reflected by the membrane. The sensor(s) is illuminated by a uniform tight field, and the reflection from the sensor yields acoustic wave amplitude and phase information which can be picked up electronically or otherwise.
Lindahl, G; Lundström, K; Tornberg, E
2001-10-01
The colour of loin, M. longissimus dorsi (LD), and ham, M. biceps femoris (BF), from pure breed Hampshire, Swedish Landrace and Swedish Yorkshire pigs was studied. The contribution of the pigment content, the myoglobin forms deoxymyoglobin (Mb), oxymyoglobin (MbO) and metmyoglobin (MetMb) and the internal reflectance to the colour of pork of normal meat quality was evaluated using partial least squares regression (PLS). The colour of LD and BF from the Hampshire breed was more red and yellow and more saturated than the colour of the same muscles from the Swedish Landrace and the Swedish Yorkshire breeds. Furthermore, BF from Hampshire was darker than BF from the other two breeds. These differences in colour were related to the lower pH in Hampshire, resulting in more blooming and in higher internal reflectance, and to the higher pigment content. The colour of BF was darker and more red than the colour of LD within each breed. No colour difference was found between gilts and castrates within each breed. Most of the variation (86-90%) in lightness (L* value), redness (a* value) and yellowness (b* value), chroma (saturation) and hue angle of pork of normal meat quality was explained by the pigment content, myoglobin forms and internal reflectance. The L* value, a* value, chroma and hue angle were influenced by both the pigment content and by the myoglobin forms to almost the same extent, while the internal reflectance was of no significance to these colour parameters. The b* value was influenced most by the myoglobin forms, less by the internal reflectance and almost not at all by the pigment content.
"Beginning with the end in mind": imagining personal retirement speeches to promote professionalism.
Yu, Eunice; Wright, Scott M
2015-06-01
The goal of teaching professionalism in medicine is to transform a theoretical concept into an internalized and actualized identity. Many trainees struggle with professionalism in the abstract, particularly when instruction methods are didactic and disconnected from personal experience. The authors aim to demonstrate the feasibility of having interns frame a personal definition of professionalism based on a reflective technique called "beginning with the end in mind." Interns composed remarks that might be used to introduce them at their own retirement ceremony following a career in medicine. This "career eulogies" exercise was introduced to groups of six interns during the first third of the internship year as part of a two-week curriculum focused on professional development. Two investigators independently coded the written introductions, identifying emergent themes through content analysis. Of the 19 interns in an internal medicine residency program (2012-13), 17 participated in the exercise. Six themes emerged from the data: aligning behaviors with core values, achieving excellence in medicine, changing the world and giving back, valuing teamwork and relationships, realizing work-life balance, and appreciating a career in medicine. These themes correlate with accepted published definitions of professionalism. The personal reflections produced through this exercise allow physicians to begin to formulate their professional self-conception. Extensions of this work might include linking such forms of critical reflection to individualized learning plans and updating the speeches over time. Further research on "reflecting forward" may determine its impact as a complement to traditional narrative reflection.
Total internal reflection laser tools and methods
Zediker, Mark S.; Faircloth, Brian O.; Kolachalam, Sharath K.; Grubb, Daryl L.
2016-02-02
There is provided high power laser tools and laser heads that utilize total internal reflection ("TIR") structures to direct the laser beam along a laser beam path within the TIR structure. The TIR structures may be a TIR prism having its hypotenuse as a TIR surface.
Amplified total internal reflection: theory, analysis, and demonstration of existence via FDTD.
Willis, Keely J; Schneider, John B; Hagness, Susan C
2008-02-04
The explanation of wave behavior upon total internal reflection from a gainy medium has defied consensus for 40 years. We examine this question using both the finite-difference time-domain (FDTD) method and theoretical analyses. FDTD simulations of a localized wave impinging on a gainy half space are based directly on Maxwell's equations and make no underlying assumptions. They reveal that amplification occurs upon total internal reflection from a gainy medium; conversely, amplification does not occur for incidence below the critical angle. Excellent agreement is obtained between the FDTD results and an analytical formulation that employs a new branch cut in the complex "propagation-constant" plane.
Panigrahi, Ritwik; Srivastava, Suneel K.
2015-01-01
In present work, spherical core (polystyrene, PS)/shell (polypyrrole, PPy) has been synthesized via in situ chemical oxidative copolymerization of pyrrole (Py) on the surface of sulfonated PS microsphere followed by the formation of hollow polypyrrole (HPPy) shell by dissolving PS inner core in THF. Thereafter, we first time established that such fabricated novel art of morphology acts as a conducting trap in absorbing electromagnetic (EM) wave by internal reflection. Further studies have been extended on the formation of its silver nanocomposites HPPy/Ag to strengthen our contention on this novel approach. Our investigations showed that electromagnetic interference (EMI) shielding efficiency (SE) of HPPy (34.5-6 dB) is significantly higher compared to PPy (20-5 dB) in the frequency range of 0.5-8 GHz due to the trapping of EM wave by internal reflection. We also observed that EMI shielding is further enhanced to 59–23 in 10 wt% Ag loaded HPPy/Ag-10. This is attributed to the simultaneous contribution of internal reflection as well as reflection from outer surface. Such high EMI shielding capacity using conducting polymers are rarely reported. PMID:25560384
ERIC Educational Resources Information Center
Foley, Tyler; Pegram, Matthew; Jenkins, Zachary; Hester, Brooke C.; Burris, Jennifer L.
2015-01-01
We have developed an eye-catching demonstration that showcases a variety of physics topics from total internal reflection to electrostatics to non-Newtonian fluid dynamics, including the Kaye effect. The essential components of the demonstration include a vertical stream of liquid soap in which a laser pointer is internally reflected, and which…
Modular off-axis solar concentrator
Plesniak, Adam P; Hall, John C
2015-01-27
A solar concentrator including a housing defining a vertical axis and including a receiving wall connected to a reflecting wall to define an internal volume and an opening into the internal volume, wherein the reflecting wall defines at least one primary optical element, and wherein at least a portion of the reflecting wall includes a layer of reflective material, the housing further including a cover connected to the receiving wall and the reflecting wall to seal the opening, and at least one receiver mounted on the receiving wall such that a vertical axis of the receiver is disposed at a non-zero angle relative to the vertical axis of the housing, the receiver including at least one photovoltaic cell.
Cylindrical microlens with an internally reflecting surface and a method of fabrication
Beach, Raymond J.; Freitas, Barry L.
2004-03-23
A fast (high numerical aperture) cylindrical microlens, which includes an internally reflective surface, that functions to deviate the direction of the light that enters the lens from its original propagation direction is employed in optically conditioning laser diodes, laser diode arrays and laser diode bars.
A Cylindrical Microlens With An Internally Reflective Surface And A Method Of Fabrication
Beach, Raymond J.; Freitas, Barry L.
2005-09-27
A fast (high numerical aperture) cylindrical microlens, which includes an internally reflective surface, that functions to deviate the direction of the light that enters the lens from its original propagation direction is employed in optically conditioning laser diodes, laser diode arrays and laser diode bars.
Resonant tunneling in frustrated total internal reflection.
Longhi, Stefano
2005-10-15
Anomalous light transmission and resonant tunneling in frustrated total internal reflection (FTIR) are theoretically predicted to occur at periodically curved interfaces. For a low-contrast index and for grazing incidence, it is shown that FTIR resonant tunneling provides an optical realization of field-induced barrier transparency in quantum tunneling.
NASA Technical Reports Server (NTRS)
Harper, L. L. (Inventor)
1983-01-01
An optical resonator cavity configuration has a unitary mirror with oppositely directed convex and concave reflective surfaces disposed into one fold and concertedly reversing both ends of a beam propagating from a laser rod disposed between two total internal reflection prisms. The optical components are rigidly positioned with perpendicularly crossed virtual rooflines by a compact optical bed. The rooflines of the internal reflection prisms, are arranged perpendicularly to the axis of the laser beam and to the optical axes of the optical resonator components.
Asokan, Sreeja B.; Johnson, Heath E.; Rahman, Anisur; King, Samantha J.; Rotty, Jeremy D.; Lebedeva, Irina P.; Haugh, Jason M.; Bear, James E.
2014-01-01
Summary Chemotaxis, migration towards soluble chemical cues, is critical for processes such as wound healing and immune surveillance, and is exhibited by various cell types from rapidly-migrating leukocytes to slow-moving mesenchymal cells. To interrogate the mechanisms involved in mesenchymal chemotaxis, we observed cell migration in microfluidic chambers that generate stable gradients of the chemoattractant PDGF. Surprisingly, we found that pathways implicated in amoeboid chemotaxis, such as PI3K and mTOR signaling, are dispensable for chemotaxis to PDGF. Instead, we find that local inactivation of Myosin IIA, through a non-canonical Ser1/2 phosphorylation of the regulatory light chain, is essential. This site is phosphorylated by PKCα, which is activated by an intracellular gradient of diacylglycerol generated by PLCγ. Using a combination of TIRF imaging and gradients of activators/inhibitors in the microfluidic chambers, we demonstrate that this signaling pathway and subsequent inhibition of Myosin II activity at the leading edge is required for mesenchymal chemotaxis. PMID:25482883
Gustafsson, Nils; Culley, Siân; Ashdown, George; Owen, Dylan M.; Pereira, Pedro Matos; Henriques, Ricardo
2016-01-01
Despite significant progress, high-speed live-cell super-resolution studies remain limited to specialized optical setups, generally requiring intense phototoxic illumination. Here, we describe a new analytical approach, super-resolution radial fluctuations (SRRF), provided as a fast graphics processing unit-enabled ImageJ plugin. In the most challenging data sets for super-resolution, such as those obtained in low-illumination live-cell imaging with GFP, we show that SRRF is generally capable of achieving resolutions better than 150 nm. Meanwhile, for data sets similar to those obtained in PALM or STORM imaging, SRRF achieves resolutions approaching those of standard single-molecule localization analysis. The broad applicability of SRRF and its performance at low signal-to-noise ratios allows super-resolution using modern widefield, confocal or TIRF microscopes with illumination orders of magnitude lower than methods such as PALM, STORM or STED. We demonstrate this by super-resolution live-cell imaging over timescales ranging from minutes to hours. PMID:27514992
3D Filament Network Segmentation with Multiple Active Contours
NASA Astrophysics Data System (ADS)
Xu, Ting; Vavylonis, Dimitrios; Huang, Xiaolei
2014-03-01
Fluorescence microscopy is frequently used to study two and three dimensional network structures formed by cytoskeletal polymer fibers such as actin filaments and microtubules. While these cytoskeletal structures are often dilute enough to allow imaging of individual filaments or bundles of them, quantitative analysis of these images is challenging. To facilitate quantitative, reproducible and objective analysis of the image data, we developed a semi-automated method to extract actin networks and retrieve their topology in 3D. Our method uses multiple Stretching Open Active Contours (SOACs) that are automatically initialized at image intensity ridges and then evolve along the centerlines of filaments in the network. SOACs can merge, stop at junctions, and reconfigure with others to allow smooth crossing at junctions of filaments. The proposed approach is generally applicable to images of curvilinear networks with low SNR. We demonstrate its potential by extracting the centerlines of synthetic meshwork images, actin networks in 2D TIRF Microscopy images, and 3D actin cable meshworks of live fission yeast cells imaged by spinning disk confocal microscopy.
Even illumination in total internal reflection fluorescence microscopy using laser light.
Fiolka, R; Belyaev, Y; Ewers, H; Stemmer, A
2008-01-01
In modern fluorescence microscopy, lasers are a widely used source of light, both for imaging in total internal reflection and epi-illumination modes. In wide-field imaging, scattering of highly coherent laser light due to imperfections in the light path typically leads to nonuniform illumination of the specimen, compromising image analysis. We report the design and construction of an objective-launch total internal reflection fluorescence microscopy system with excellent evenness of specimen illumination achieved by azimuthal rotation of the incoming illuminating laser beam. The system allows quick and precise changes of the incidence angle of the laser beam and thus can also be used in an epifluorescence mode. 2007 Wiley-Liss, Inc
"Once when i was on call...," theory versus reality in training for professionalism.
Eggly, Susan; Brennan, Simone; Wiese-Rometsch, Wilhelmine
2005-04-01
To identify the degree to which interns' reported experiences with professional and unprofessional behavior converge and/or diverge with ideal professional behavior proposed by the physician community. Interns at Wayne State University's residency programs in internal medicine, family medicine, and transitional medicine responded to essay questions about their experience with professional and unprofessional behavior as part of a curriculum on professionalism. Responses were coded for whether they reflected each of the principles and responsibilities outlined in a major publication on physician professionalism. Content analysis included the frequencies with which the interns' essays reflected each principle or responsibility. Additionally, a thematic analysis revealed themes of professional behavior that emerged from the essays. Interns' experiences with professional and unprofessional behavior most frequently converged with ideal behavior proposed by the physician community in categories involving interpersonal interactions with patients. Interns infrequently reported experiences involving behavior related to systems or sociopolitical issues. Interns' essays reflect their concern with interpersonal interactions with patients, but they are either less exposed to or less interested in describing behavior regarding systems or sociopolitical issues. This may be due to their stage of training or to the emphasis placed on interpersonal rather than systems or sociopolitical issues during training. The authors recommend future proposals of ideal professional behavior be revised periodically to reflect current experiences of practicing physicians, trainees, other health care providers and patients. Greater educational emphasis should be placed on the systems and sociopolitical environment in which trainees practice.
ERIC Educational Resources Information Center
Bitz, Michael; Emejulu, Obiajulu
2016-01-01
This article is an international reflection on literacy, creativity, and student engagement. The authors collaborated to help Nigerian youths and their teachers develop, design, and share original comic books. By leveraging student engagement for literacy learning, the authors highlighted the crucial role of creativity in the classroom. The…
Students' Quality of Mathematical Discussion and Their Self-Determination in Mathematics
ERIC Educational Resources Information Center
Kosko, Karl W.; Wilkins, Jesse L. M.
2012-01-01
Mathematical discussion allows for students to reflect upon math concepts and understand such concepts at a deeper level. This process of reflection requires a certain amount of internalization on the part of the student. This internalization is facilitated by meeting the needs of autonomy, competence, and relatedness as advocated by…
ERIC Educational Resources Information Center
Saa'd AlDin, Kawther
2014-01-01
In 2010, the International Baccalaureate (IB) Organization mandated that all its schools, including Diploma (DP) schools, adhere to the collaborative planning and reflection requirements, which emphasized the importance of integrating its theory of knowledge (TOK) core component into all disciplines. Many schools officials and educations in Amman…
ERIC Educational Resources Information Center
Clark, J. Spencer; Brown, James Scott; Jandildinov, Medet
2016-01-01
The concepts of reflection and reflective practice have become the core of many teacher education programmes, with critical reflection as the goal for many teacher educators. This study examined the use of a videoconference discussion in an instructional methodology course as a means to enrich the process of reflection and encourage critical…
Where is the "meta" in animal metacognition?
Kornell, Nate
2014-05-01
Apes, dolphins, and some monkeys seem to have metacognitive abilities: They can accurately evaluate the likelihood that their response in cognitive task was (or will be) correct. These certainty judgments are seen as significant because they imply that animals can evaluate internal cognitive states, which may entail meaningful self-reflection. But little research has investigated what is being reflected upon: Researchers have assumed that when animals make metacognitive judgments they evaluate internal memory strength. Yet decades of research have demonstrated that humans cannot directly evaluate internal memory strength. Instead, they make certainty judgments by drawing inferences from cues they can evaluate, such as familiarity and ease of processing. It seems likely that animals do the same, but this hypothesis has not been tested. I suggest two strategies for investigating the internal cues that underlie animal metacognitive judgments. It is possible that animals, like humans, are capable of making certainty judgments based on internal cues without awareness or meaningful self-reflection. ©2014 APA, all rights reserved.
NASA Astrophysics Data System (ADS)
Fontaine, Norman Henry
1997-10-01
Techniques which can be used to obtain depth-resolved information on the thermodynamics at polymer-polymer and polymer-wall interfaces, and of small molecule diffusion in polymers, are of particular interest to industry. Optical methods which are sensitive to molecular vibrations (such as internal reflection Raman spectroscopy) are advantageous because they can non- destructively probe molecular content, orientation, and polarity of the local environment in a sample. However, while optical internal reflection depth-profiling methods have been reported, they have never progressed beyond the demonstration stage. In this work, the theory and methodology of internal reflection spectroscopy are developed and optimized into a rigorous field-controlled spectroscopic technique. A novel asymmetric internal reflection element (IRE) is introduced which traps back-reflections, allowing precise evanescent and standing wave probe-field control in the sample for all angles of incidence. It is demonstrated that a Gaussian laser beam will best approximate an infinite homogeneous plane wave when the IRE/sample interface lies in the paraxial-Fraunhofer region (far- field) of the beam path. Calibration methods are presented, sources of systematic errors are identified, and the angular resolution limit (ARL) is introduced as a measure of the field control developed in a sample by any internal reflection method. A general model of Raman scattering and photon detection from multi-layer thin films is developed. A new and generalized operator based transfer matrix method is developed and applied to electromagnetic field and diffusion computations in multi-layer systems. Total internal reflection spectroscopy is extended to include sub-critical angles of incidence, where resonant field enhancements generate large and selective amplification of the probe-field intensity within the layers of the sample. Fitting these resonances to the model spectral intensities allows unique determination of the location of buried interfaces in micron-sized polymer multi-layers with nanometer scale precision and the refractive indices of the layers with precision of /Delta n/approx/pm 0.0001. The Raman active molecular content of each optically distinct layer of the film is determinable simultaneously with the optical properties. Resonant mode VAIRRS studies of poly(methyl methacrylate) films spun-cast from toluene and then dried under ambient conditions have shown evidence for toluene diffusion concurrent with a rotationally hindered relaxation of oriented ester side groups about the polymer backbone. Low temperature annealing (≈87oC) has shown evidence that this hindered rotational relaxation may be reversible. VAIRRS study of a polystyrene/poly(methyl methacrylate) bi-layer has detected evidence for toluene diffusion across the buried polymer-polymer interface.
NASA Astrophysics Data System (ADS)
Kao, I.-Ling; Ku, Chun-Neng; Chen, Yi-Ping; Lin, Ding-Zheng
2012-09-01
We proposed an internal nanostructure with a high reflective index planarization layer to solve the optical loss due to the reflective index mismatch between ITO and glass substrate. In our experiments, we found the electrical property of OLED device was significantly influenced by the internal nanostructures without planarization layer. Moreover, the internal extraction structure (IES) is not necessarily beneficial for light extraction. Therefore, we proposed a new substrate combine both internal and external extraction structure (EES) to extract trapping light. We successfully developed a high refractive index (N 1.7) planarization material with flat surface (RMS roughness < 2 nm), and improved about 70% device efficiency compared to traditional glass substrate.
Nonimaging light concentrator with uniform irradiance
Winston, Roland; Gee, Randy C.
2003-04-01
A nonimaging light concentrator system including a primary collector of light, an optical mixer disposed near the focal zone for collecting light from the primary collector, the optical mixer having a transparent entrance aperture, an internally reflective housing for substantially total internal reflection of light, a transparent exit aperture and an array of photovoltaic cells disposed near the transparent exit aperture.
Reflections on Building "Glocal" Competence among Pre-Service and In-Service Teachers
ERIC Educational Resources Information Center
Gichiru, Wangari
2016-01-01
The purpose of this paper is to critically reflect on two graduate international comparative education courses I taught at a mid-sized public university. Using a variety of readings and multimedia focusing on voices of international educators telling their own stories of struggle for a democratic education, these courses represent my effort to…
ERIC Educational Resources Information Center
King, Virginia; Clouder, Lynn; Deane, Mary; Deepwell, Frances; Ganobcsik-Williams, Lisa
2007-01-01
Through reflection on the First International iPED Conference 2006, and its overarching theme of "Pedagogical Research and Academic Identities", this paper considers the achievement of the wider aims of the conference, which were to facilitate dialogue between researchers in order to explore the conference themes collaboratively, and to…
Symbolic Uses of Evaluation in the International Aid Sector: Arguments for Critical Reflection
ERIC Educational Resources Information Center
McNulty, James
2012-01-01
Significant progress has been made in recent years to improve the quality of the evaluation of international aid. Increasingly, this includes an interest in improving the way evaluations are used to improve policies and programmes. However, the prevalence of symbolic use--a phenomenon that is often mentioned but rarely studied--reflects an…
Solar module having reflector between cells
Kardauskas, Michael J.
1999-01-01
A photovoltaic module comprising an array of electrically interconnected photovoltaic cells disposed in a planar and mutually spaced relationship between a light-transparent front cover member in sheet form and a back sheet structure is provided with a novel light-reflecting means disposed between adjacent cells for reflecting light falling in the areas between cells back toward said transparent cover member for further internal reflection onto the solar cells. The light-reflecting comprises a flexible plastic film that has been embossed so as to have a plurality of small V-shaped grooves in its front surface, and a thin light-reflecting coating on said front surface, the portions of said coating along the sides of said grooves forming light-reflecting facets, said grooves being formed so that said facets will reflect light impinging thereon back into said transparent cover sheet with an angle of incidence greater than the critical angle, whereby substantially all of the reflected light will be internally reflected from said cover sheet back to said solar modules, thereby increasing the current output of the module.
ERIC Educational Resources Information Center
Ashton, Karen
2016-01-01
This paper reflects on the methodology used in international comparative education surveys by conducting a systematic review of the European Survey on Language Competences (ESLC). The ESLC was administered from February to March 2011, with final results released in June 2012. The survey tested approximately 55,000 students across 14 European…
Revisit: A Surprising Demonstration of Total Internal Reflection
ERIC Educational Resources Information Center
Lee, Jiwon; Cha, Yu Wha; Jung, Yeon Su; Oh, Eun Ju; Moon, Ye Lin; Kim, Jung Bog
2016-01-01
Melton demonstrated a surprising disappearance using total internal reflection. When he put a Florence flask filled with marbles into a water tank and looked straight down from directly above the flask, he was only able to see marbles above a certain water level. When he added more water into the tank above the top line of the marbles, all of the…
ERIC Educational Resources Information Center
Fenton-Smith, Ben; Michael, Rowan
2013-01-01
This paper evaluates a strategy to promote the uptake of support services by international students (ISs) at an Australian university. As part of their assessment, ISs completed a so-called "University Service Reflection Task" (USRT) in a core first-year course. To complete the USRT, all ISs accessed one support service (e.g. language…
ERIC Educational Resources Information Center
Ng, Vinci; Yuen, Mantak
2015-01-01
This paper reflects upon the relevance of Buddhism to counselling in general and to career counseling in particular by discussing a program implemented at an international school in Hong Kong. The authors provide an analysis of the pertinent literature related to relevant concepts within Buddhism. This topic has not yet been adequately researched…
Reflections on the 12th International Transformative Learning Conference
ERIC Educational Resources Information Center
Schapiro, Steven A.; Gallegos, Placida V.; Stashower, Keren; Clark, Donna F.
2017-01-01
This article is a reflective essay that explores the question: What can the content and experience of the conference tell us about the state of theory and practice in the field of TL; where is it today and where it may be going in the future? The 12th International Transformative Learning Conference (ITLC) held October 19-23 at Pacific Lutheran…
Reflective Writing through the Use of Guiding Questions
ERIC Educational Resources Information Center
Moussa-Inaty, Jase
2015-01-01
Reflections can be seen as powerful tools for growth and intellectual development. It is no surprise that the writing of reflections is common practice at a Federal Institute in the United Arab Emirates (UAE). The research presented sought to explore possible differences in reflective writing once guidelines were presented to a group of interns in…
Total internal reflection optical switch using the reverse breakdown of a pn junction in silicon.
Kim, Jong-Hun; Park, Hyo-Hoon
2015-11-01
We demonstrate a new type of silicon total-internal-reflection optical switch with a simple pn junction functioning both as a reflector and a heater. The reflector is placed between asymmetrically y-branched multimode waveguides with an inclination angle corresponding to half of the branch angle. When the reflector is at rest, incident light is reflected in accordance to the refractive index difference due to the plasma dispersion effect of the pre-doped carriers. Switching to the transmission state is attained under a reverse breakdown of the pn junction by the thermo-optic effect which smears the refractive index difference. From this switching scheme, we confirmed the switching operation with a shallow total-internal-reflection region of 1 μm width. At a 6° branch angle, an extinction ratio of 12 dB and an insertion loss of -4.2 dB are achieved along with a thermal heating power of 151.5 mW.
Zhang, Jiwei; Di, Jianglei; Li, Ying; Xi, Teli; Zhao, Jianlin
2015-10-19
We present a method for dynamically measuring the refractive index distribution in a large range based on the combination of digital holographic interferometry and total internal reflection. A series of holograms, carrying the index information of mixed liquids adhered on a total reflection prism surface, are recorded with CCD during the diffusion process. Phase shift differences of the reflected light are reconstructed exploiting the principle of double-exposure holographic interferometry. According to the relationship between the reflection phase shift difference and the liquid index, two dimensional index distributions can be directly figured out, assuming that the index of air near the prism surface is constant. The proposed method can also be applied to measure the index of solid media and monitor the index variation during some chemical reaction processes.
Electrowetting-actuated optical switch based on total internal reflection.
Liu, Chao; Wang, Di; Yao, Li-Xiao; Li, Lei; Wang, Qiong-Hua
2015-04-01
In this paper we demonstrate a liquid optical switch based on total internal reflection. Two indium tin oxide electrodes are fabricated on the bottom substrate. A conductive liquid (Liquid 1) is placed on one side of the chamber and surrounded by a density-matched silicone oil (Liquid 2). In initial state, when the light beam illuminates the interface of the two liquids, it just meets the conditions of total internal reflection. The light is totally reflected by Liquid 2, and the device shows light-off state. When we apply a voltage to the other side of the indium tin oxide electrode, Liquid 1 stretched towards this side of the substrate and the curvature of the liquid-liquid interface changes. The light beam is refracted by Liquid 1 and the device shows light-on state. So the device can achieve the functions of an optical switch. Because the light beam can be totally reflected by the liquid, the device can attain 100% light intensity attenuation. Our experiments show that the response time from light-on (off) to light-off (on) are 130 and 132 ms, respectively. The proposed optical switch has potential applications in variable optical attenuators, information displays, and light shutters.
Diffractive flat panel solar concentrators of a novel design.
de Jong, Ties M; de Boer, Dick K G; Bastiaansen, Cees W M
2016-07-11
A novel design for a flat panel solar concentrator is presented which is based on a light guide with a grating applied on top that diffracts light into total internal reflection. By combining geometrical and diffractive optics the geometrical concentration ratio is optimized according to the principles of nonimaging optics, while the thickness of the device is minimized due to the use of total internal reflection.
Apparent bandgap shift in the internal quantum efficiency for solar cells with back reflectors
NASA Astrophysics Data System (ADS)
Steiner, M. A.; Perl, E. E.; Geisz, J. F.; Friedman, D. J.; Jain, N.; Levi, D.; Horner, G.
2017-04-01
We demonstrate that in solar cells with highly reflective back mirrors, the measured internal quantum efficiency exhibits a shift in bandgap relative to the measured external quantum efficiency. The shift arises from the fact that the measured reflectance at the front surface includes a superposition of waves reflecting from the front and back surfaces. We quantify the magnitude of the apparent shift and discuss the errors that can result in determination of quantities such as the photocurrent. Because of this apparent shift, it is important the bandgap be determined from the external quantum efficiency.
Apparent bandgap shift in the internal quantum efficiency for solar cells with back reflectors
Steiner, Myles A.; Perl, E. E.; Geisz, J. F.; ...
2017-04-28
Here, we demonstrate that in solar cells with highly reflective back mirrors, the measured internal quantum efficiency exhibits a shift in bandgap relative to the measured external quantum efficiency. The shift arises from the fact that the measured reflectance at the front surface includes a superposition of waves reflecting from the front and back surfaces. We quantify the magnitude of the apparent shift and discuss the errors that can result in determination of quantities such as the photocurrent. Because of this apparent shift, it is important that the bandgap be determined from the external quantum efficiency.
Lepp, Margret; Zorn, CeCelia R; Duffy, Patricia R; Dickson, Rana J
2003-01-01
Reflection, a process grounded in distancing from the self to enhance self-awareness, can be used as a pedagogic activity to promote students' transition to a greater authenticity and professionalism and, therefore, improve patient care and nursing practice. In this international educational project (implemented in 2001) using interactive videoconferencing technology (IVC), Swedish and U.S. nursing students and faculty incorporated reflective journaling, drama in education, photolanguage, and off-air meeting discussions to enhance personal and professional development. Highlights from the literature, a description of the project, and student and faculty appraisals are presented.
Total Internal Reflection Accounts for the Bright Color of the Saharan Silver Ant
Aron, Serge
2016-01-01
The Saharan silver ant Cataglyphis bombycina is one of the terrestrial living organisms best adapted to tolerate high temperatures. It has recently been shown that the hairs covering the ant’s dorsal body part are responsible for its silvery appearance. The hairs have a triangular cross-section with two corrugated surfaces allowing a high optical reflection in the visible and near-infrared (NIR) range of the spectrum while maximizing heat emissivity in the mid-infrared (MIR). Those two effects account for remarkable thermoregulatory properties, enabling the ant to maintain a lower thermal steady state and to cope with the high temperature of its natural habitat. In this paper, we further investigate how geometrical optical and high reflection properties account for the bright silver color of C. bombycina. Using optical ray-tracing models and attenuated total reflection (ATR) experiments, we show that, for a large range of incidence angles, total internal reflection (TIR) conditions are satisfied on the basal face of each hair for light entering and exiting through its upper faces. The reflection properties of the hairs are further enhanced by the presence of the corrugated surface, giving them an almost total specular reflectance for most incidence angles. We also show that hairs provide an almost 10-fold increase in light reflection, and we confirm experimentally that they are responsible for a lower internal body temperature under incident sunlight. Overall, this study improves our understanding of the optical mechanisms responsible for the silver color of C. bombycina and the remarkable thermoregulatory properties of the hair coat covering the ant’s body. PMID:27073923
NASA Technical Reports Server (NTRS)
Cavitte, Marie G. P.; Blankenship, Donald D.; Young, Duncan A.; Schroeder, Dustin M.; Parrenin, Frederic; Lemeur, Emmanuel; Macgregor, Joseph A.; Siegert, Martin J.
2016-01-01
Several airborne radar-sounding surveys are used to trace internal reflections around the European Project for Ice Coring in Antarctica Dome C and Vostok ice core sites. Thirteen reflections, spanning the last two glacial cycles, are traced within 200 km of Dome C, a promising region for million-year-old ice, using the University of Texas Institute for Geophysics High-Capacity Radar Sounder. This provides a dated stratigraphy to 2318 m depth at Dome C. Reflection age uncertainties are calculated from the radar range precision and signal-to-noise ratio of the internal reflections. The radar stratigraphy matches well with the Multichannel Coherent Radar Depth Sounder (MCoRDS) radar stratigraphy obtained independently. We show that radar sounding enables the extension of ice core ages through the ice sheet with an additional radar-related age uncertainty of approximately 1/3-1/2 that of the ice cores. Reflections are extended along the Byrd-Totten Glacier divide, using University of Texas/Technical University of Denmark and MCoRDS surveys. However, core-to-core connection is impeded by pervasive aeolian terranes, and Lake Vostok's influence on reflection geometry. Poor radar connection of the two ice cores is attributed to these effects and suboptimal survey design in affected areas. We demonstrate that, while ice sheet internal radar reflections are generally isochronal and can be mapped over large distances, careful survey planning is necessary to extend ice core chronologies to distant regions of the East Antarctic ice sheet.
Exploring Dimensions of Critical Reflection in Activist-Facilitator Practice
ERIC Educational Resources Information Center
Hanson, Cindy
2013-01-01
This article explores how 14 diverse, Canadian activist-facilitators working in international development experience and understand "critical reflection" as a component of participatory methodologies in facilitation practices. The findings, based on my doctoral study, demonstrate that although critical reflection is often discussed as…
Reilly, Thomas
2008-02-15
The history of the Journal of Sports Sciences is traced from the antecedents of its initiation to the current time. The developments of the sports sciences at large are reflected in the content of the journal. Its links with the international agenda are described, and related to landmark publications. Special attention is given to the relationships with international bodies, the International Society for Advancement of Kinanthropometry and the World Commission of Science and Sports. The expansion of sport and exercise sciences, both nationally and internationally, was reflected in the increased frequency of publication of the journal. Key areas in the kinanthropometric content are identified and placed in context. The review culminates in the highlighting of likely areas for future research.
Refractive index measurement for biomaterial samples by total internal reflection.
Jin, Y L; Chen, J Y; Xu, L; Wang, P N
2006-10-21
The refractive index of biological tissue is a fundamental parameter in applications of optical diagnosis and laser treatments. In the present work, the refractive indices and thermo-optic coefficients of some basic biomaterials, such as blood plasma, haemoglobin solution and lipid membrane, were studied by the method of total internal reflection at the wavelengths of 532 and 632.8 nm that are the most frequently used laser wavelengths in the biomedical field. The effects of the sample concentration and the temperature on refractive index were measured, and empirical relationships were summarized, accompanied by a theoretical explanation based on molecular polarization theory. The results provide some fundamental data for the refractive indices of these biomaterials under variant conditions, and also demonstrate that the total internal reflection method is a feasible and reliable way to measure the refractive indices of biological samples.
Monlithic nonplanar ring oscillator and method
NASA Technical Reports Server (NTRS)
Nilsson, Alan C. (Inventor); Byer, Robert L. (Inventor)
1991-01-01
A monolithic nonplanar ring oscillator having an optically isotropic solid-state laser body for propagating laser radiation about a nonplanar ring path internal to the laser body is disclosed. The monolithic laser body is configured to produce a 2N reflection nonplanar ring light path, where N is an integer greater than or equal to 2, comprising 2N-1 total internal reflections and one reflection at a coupler in a single round trip. Undirectional traveling wave oscillation of the laser is induced by the geometry of the nonplanar ring path together with the effect of an applied magnetic field and partial polarizer characteristics of the oblique reflection from the coupler. The 6-reflection nonplanar ring oscillator makes possible otpimal unidirectional oscillation (low loss for the oscillating direction of propagation and, simultaneously high loss for the nonoscillating direction of propagation) in monolithic NPROs using materials with index of refraction smaller than the square root of 3, for example, laser glass.
NASA Astrophysics Data System (ADS)
Dickinson, Alex; White, N. J.; Caulfield, C. P.
2017-12-01
Bright reflections are observed within the upper 1,000 m of the water column along a seismic reflection profile that traverses the northern margin of the Gulf of Mexico. Independent hydrographic calibration demonstrates that these reflections are primarily caused by temperature changes associated with different water masses that are entrained into the Gulf along the Loop Current. The internal wave field is analyzed by automatically tracking 1,171 reflections, each of which is greater than 2 km in length. Power spectra of the horizontal gradient of isopycnal displacement, ϕξx, are calculated from these tracked reflections. At low horizontal wave numbers (kx<10-2 cpm), ϕξx∝kx-0.2±0.6, in agreement with hydrographic observations of the internal wave field. The turbulent spectral subrange is rarely observed. Diapycnal diffusivity, K, is estimated from the observed internal wave spectral subrange of each tracked reflection using a fine-scale parametrization of turbulent mixing. Calculated values of K vary between 10-8 and 10-4 m2 s-1 with a mean value of K˜4×10-6 m2 s-1. The spatial distribution of turbulent mixing shows that K˜10-7 m2 s-1 away from the shelf edge in the upper 300 m where stratification is strong. Mixing is enhanced by up to 4 orders of magnitude adjacent to the shoaling bathymetry of the continental slope. This overall pattern matches that determined by analyzing nearby suites of CTD casts. However, the range of values recovered by spectral analysis of the seismic image is greater as a consequence of significantly better horizontal resolution.
Lamagna, Jerry; Gleiser, Kari A
2007-01-01
In this paper, we introduce Intra-Relational AEDP (I-R) as an attachment-based experiential approach to trauma treatment. Integrating Accelerated Experiential Dynamic Psychotherapy (AEDP) (Fosha, 2000a,b, 2002, 2003) with ego-state methodology, intra-relational interventions specifically seeks to help clients by (1) fostering capacities for self-regulation through shared states of affective resonance between therapist, client, and dissociated self-states; (2) facilitating authentic, open internal dialogue between self-states which can alter engrained patterns of intra-psychic conflict and self-punishment; (3) developing abilities for self-reflection and emotional processing by co-mingling previously disowned affect and emotional memories with here and now experience; and (4) attending to positive affects evoked through experiences of transformation, self-compassion, and self-affirmation. Drawing from object relations and attachment theory, intra-relational interventions places particular emphasis on internal attachment relationships formed through interactions between the client's subjective selves (internal subjects) and reflective selves (internal objects). Through visual imagery, internal dialogue, and explicit relational techniques, intra-relational interventions aims to develop this subjective-reflective dyad's capacity for reciprocal attunement, resonance, and responsiveness. Such clinical strategies aim to foster healing and psychological integration between the client and heretofore disavowed aspects of self.
Patterson, Brian M; Havrilla, George J
2006-11-01
The number of techniques and instruments available for Fourier transform infrared (FT-IR) microspectroscopic imaging has grown significantly over the past few years. Attenuated total internal reflectance (ATR) FT-IR microspectroscopy reduces sample preparation time and has simplified the analysis of many difficult samples. FT-IR imaging has become a powerful analytical tool using either a focal plane array or a linear array detector, especially when coupled with a chemometric analysis package. The field of view of the ATR-IR microspectroscopic imaging area can be greatly increased from 300 x 300 microm to 2500 x 2500 microm using a larger internal reflection element of 12.5 mm radius instead of the typical 1.5 mm radius. This gives an area increase of 70x before aberrant effects become too great. Parameters evaluated include the change in penetration depth as a function of beam displacement, measurements of the active area, magnification factor, and change in spatial resolution over the imaging area. Drawbacks such as large file size will also be discussed. This technique has been successfully applied to the FT-IR imaging of polydimethylsiloxane foam cross-sections, latent human fingerprints, and a model inorganic mixture, which demonstrates the usefulness of the method for pharmaceuticals.
Culturing In Vivo-like Murine Astrocytes Using the Fast, Simple, and Inexpensive AWESAM Protocol.
Wolfes, Anne C; Dean, Camin
2018-01-10
The AWESAM (a low-cost easy stellate astrocyte method) protocol entails a fast, simple, and inexpensive way to generate large quantities of in vivo-like mouse and rat astrocyte monocultures: Brain cells can be isolated from different brain regions, and after a week of cell culture, non-astrocytic cells are shaken off by placing the culture dishes on a shaker for 6 h in the incubator. The remaining astrocytes are then passaged into new plates with an astrocyte-specific medium (termed NB+H). NB+H contains low concentrations of heparin-binding EGF-like growth factor (HBEGF), which is used in place of serum in medium. After growing in NB+H, AWESAM astrocytes have a stellate morphology and feature fine processes. Moreover, these astrocytes have more in vivo-like gene expression than astrocytes generated by previously published methods. Ca 2+ imaging, vesicle dynamics, and other events close to the membrane can thus be studied in the fine astrocytic processes in vitro, e.g., using live cell confocal or TIRF microscopy. Notably, AWESAM astrocytes also exhibit spontaneous Ca 2+ signaling similar to astrocytes in vivo.
Raab, Mario; Jusuk, Ija; Molle, Julia; Buhr, Egbert; Bodermann, Bernd; Bergmann, Detlef; Bosse, Harald; Tinnefeld, Philip
2018-01-29
In recent years, DNA origami nanorulers for superresolution (SR) fluorescence microscopy have been developed from fundamental proof-of-principle experiments to commercially available test structures. The self-assembled nanostructures allow placing a defined number of fluorescent dye molecules in defined geometries in the nanometer range. Besides the unprecedented control over matter on the nanoscale, robust DNA origami nanorulers are reproducibly obtained in high yields. The distances between their fluorescent marks can be easily analysed yielding intermark distance histograms from many identical structures. Thus, DNA origami nanorulers have become excellent reference and training structures for superresolution microscopy. In this work, we go one step further and develop a calibration process for the measured distances between the fluorescent marks on DNA origami nanorulers. The superresolution technique DNA-PAINT is used to achieve nanometrological traceability of nanoruler distances following the guide to the expression of uncertainty in measurement (GUM). We further show two examples how these nanorulers are used to evaluate the performance of TIRF microscopes that are capable of single-molecule localization microscopy (SMLM).
Stratton, Margaret; Lee, Il-Hyung; Bhattacharyya, Moitrayee; Christensen, Sune M; Chao, Luke H; Schulman, Howard; Groves, Jay T; Kuriyan, John
2014-01-01
The activation of the dodecameric Ca2+/calmodulin dependent kinase II (CaMKII) holoenzyme is critical for memory formation. We now report that CaMKII has a remarkable property, which is that activation of the holoenzyme triggers the exchange of subunits between holoenzymes, including unactivated ones, enabling the calcium-independent phosphorylation of new subunits. We show, using a single-molecule TIRF microscopy technique, that the exchange process is triggered by the activation of CaMKII, and that exchange is modulated by phosphorylation of two residues in the calmodulin-binding segment, Thr 305 and Thr 306. Based on these results, and on the analysis of molecular dynamics simulations, we suggest that the phosphorylated regulatory segment of CaMKII interacts with the central hub of the holoenzyme and weakens its integrity, thereby promoting exchange. Our results have implications for an earlier idea that subunit exchange in CaMKII may have relevance for information storage resulting from brief coincident stimuli during neuronal signaling. DOI: http://dx.doi.org/10.7554/eLife.01610.001 PMID:24473075
ERIC Educational Resources Information Center
Campbell, Heather; Strawser, Michael G.; George, Stephen
2016-01-01
As the movement toward international education continues, institutions must be proficient when reaching and teaching international students. Instructors should engage unique learning styles, address individual student needs, and take part in additional training to effectively teach international student learners. These instructional imperatives…
Resistance and Disidentification in Reflective Practice with Preservice Teaching Interns
ERIC Educational Resources Information Center
Middleton, Michael; Abrams, Eleanor; Seaman, Jayson
2011-01-01
Teachers who engage in reflective practices are better able to recognize the complexity of teaching, use judgment to choose appropriate strategies for teaching and learning in their specific contexts, and experience improved self-confidence. For these reasons, many teacher education programs emphasize self-reflective practice. Self-reflective…
Beyond Reflection through an Academic Lens: Refraction and International Experiential Education
ERIC Educational Resources Information Center
Pagano, Monica; Roselle, Laura
2009-01-01
Students today are becoming more interested in international opportunities for study, and are drawn to alternative programs such as international service learning and international internships. These programs, however, must be carefully designed. In this paper, the authors propose using tools that go beyond the traditional understanding of…
Oreopoulos, John; Yip, Christopher M.
2009-01-01
Determining the local structure, dynamics, and conformational requirements for protein-protein and protein-lipid interactions in membranes is critical to understanding biological processes ranging from signaling to the translocating and membranolytic action of antimicrobial peptides. We report here the application of a combined polarized total internal reflection fluorescence microscopy-in situ atomic force microscopy platform. This platform's ability to image membrane orientational order was demonstrated on DOPC/DSPC/cholesterol model membranes containing the fluorescent membrane probe, DiI-C20 or BODIPY-PC. Spatially resolved order parameters and fluorophore tilt angles extracted from the polarized total internal reflection fluorescence microscopy images were in good agreement with the topographical details resolved by in situ atomic force microscopy, portending use of this technique for high-resolution characterization of membrane domain structures and peptide-membrane interactions. PMID:19254557
A Feasibility Study on Generation of Acoustic Waves Utilizing Evanescent Light
NASA Astrophysics Data System (ADS)
Matsuya, I.; Matozaki, K.; Kosugi, A.; Ihara, I.
2014-06-01
A new approach of generating acoustic waves utilizing evanescent light is presented. The evanescent light is a non-propagating electromagnetic wave that exhibits exponential decay with distance from the surface at which the total internal reflection of light is formed. In this research, the evanescent light during total internal reflection at prism surface is utilized for generating acoustic waves in aluminium and the feasibility for ultrasonic measurements is discussed. Pulsed Nd:YAG laser with 0.36 J/cm2 power density is used and the incident angle during the total internal reflection is arranged to be 69.0° for generating the evanescent light. It has been demonstrated that the amplitude of the acoustic waves by means of evanescent light is about 1/14 as large as the one generated by the conventional pulsed laser. This reveals the possibility of using a laser ultrasonic technique with near-field optics.
Bioethical Dimensions of International Cooperation for Health: still a controversial issue?
Santana, José Paranaguá de
2017-07-01
This essay stresses the importance of undertaking a critical analysis of international cooperation for health based upon reflections on diplomacy, acknowledging the ethical limits and possibilities of cooperation within this context. It emphasizes the importance of adopting an historical perspective, highlighting the circumstances surrounding the consolidation of the current international system, whereby, after two world wars, the victorious countries created the United Nations Organization, which today brings together all the world's nations, notwithstanding the fact that the majority of countries are in favor of significant changes to its institutional framework. It also stresses the importance of the bioethics dimension, despite the systematic disregard of principles adopted in relevant UN documents, especially in view of inequalities pari passu with asymmetries of knowledge and enjoyment of innovations that separate nations. This reflection also suggests that South-South cooperation is fundamental to the analysis of the inequality and dependence that demarcate North-South and East-West bipolarity. These reflections are, in short, important guiding elements for deepening analysis and resolutions in the arena of international cooperation for health, leaving aside catastrophic visions and idealistic illusions.
Touch Sensor Responds to Contact Pressure
NASA Technical Reports Server (NTRS)
Bejczy, A. K.
1982-01-01
Optical tactile sensor for mechanical hands senses contact pressure via change in light reflected from an elastic covering. Pressure against a cell cover causes distortion, which changes internal reflection of light. Change is sensed by detector, and output signal informs operator of contact. The greater the pressure and distortion, the greater the change in light reflection.
ERIC Educational Resources Information Center
Lamm, Alexa J.; Cannon, Karen J.; Roberts, T. Grady; Irani, Tracy A.; Snyder, Lori J. Unruh; Brendemuhl, Joel; Rodriguez, Mary T.
2011-01-01
Experiential learning techniques have been used in agricultural education programs for decades. An essential part of this experiential learning process is reflection. Kolb (1984) stated "knowledge results from the combination of grasping experience and transforming it" (p. 41) in a process that involves using reflection. Few researchers…
ERIC Educational Resources Information Center
Lepp, Margret; Zorn, CeCelia R.; Duffy, Patricia R.; Dickson, Rana J.
2003-01-01
A nursing course connected U.S. and Swedish sites via interactive videoconferencing and used reflective methods (journaling, drama, photo language) and off-air group discussion. Evaluation by five Swedish and seven U.S. students suggested how reflection moved students toward greater authenticity and professionalism in nursing practice. (Contains…
NASA Astrophysics Data System (ADS)
Cadatal-Raduban, Marilou; Pham, Minh Hong; Pham, Duong Van; Bui, Duong Thi Thuy; Yamanoi, Kohei; Takeda, Kohei; Empizo, Melvin John F.; Mui, Luong Viet; Shimizu, Toshihiko; Nguyen, Hung Dai; Sarukura, Nobuhiko; Fukuda, Tsuguo
2018-06-01
A two-side-pumping scheme that is based on total internal reflection in a diamond-cut Ce3+:LiCaAlF6 crystal suitable for the development of an ultraviolet laser and femtosecond amplifier system is proposed. Experimental fluorescence images and lasing results that demonstrate total internal reflection of the excitation beam using this diamond-cut crystal are presented. Calculations for the optimized crystal geometry that facilitate high extraction efficiency and homogeneity of the absorbed excitation beam are also discussed. About 50% increase in extraction efficiency compared to previously reported chirped-pulse femtosecond ultraviolet amplifier operating at 50-GW peak power is expected using this total internal reflection-based two-side-pumping configuration and a diamond-cut Ce3+:LiCaAlF6 crystal with a geometry of {φ _1} = 103°, {φ _2} = {φ _4} = 82°, {φ _3} = 93°, a length of 1.23 cm, a height of 2 cm, and an absorption coefficient of 1.5 cm-1. Our results can be used as a guide during the crystal growth process by providing the appropriate crystal geometry and size for a particular absorption coefficient to achieve high extraction efficiency. With the appropriate crystal combined with multiple-beam pumping afforded by the side-pumping scheme, the development of an all-solid-state ultraviolet laser operating at terawatt level would be within reach.
Analysis of multiple internal reflections in a parallel aligned liquid crystal on silicon SLM.
Martínez, José Luis; Moreno, Ignacio; del Mar Sánchez-López, María; Vargas, Asticio; García-Martínez, Pascuala
2014-10-20
Multiple internal reflection effects on the optical modulation of a commercial reflective parallel-aligned liquid-crystal on silicon (PAL-LCoS) spatial light modulator (SLM) are analyzed. The display is illuminated with different wavelengths and different angles of incidence. Non-negligible Fabry-Perot (FP) effect is observed due to the sandwiched LC layer structure. A simplified physical model that quantitatively accounts for the observed phenomena is proposed. It is shown how the expected pure phase modulation response is substantially modified in the following aspects: 1) a coupled amplitude modulation, 2) a non-linear behavior of the phase modulation, 3) some amount of unmodulated light, and 4) a reduction of the effective phase modulation as the angle of incidence increases. Finally, it is shown that multiple reflections can be useful since the effect of a displayed diffraction grating is doubled on a beam that is reflected twice through the LC layer, thus rendering gratings with doubled phase modulation depth.
NASA Astrophysics Data System (ADS)
Williams, Christopher
2000-07-01
The nature of international education as a field of studies has been affected by global changes over the past decade. At the same time, the concept of global security has emerged, bringing together studies related to development, the environment and the understanding of violence. Although much of the education literature reflects the global security approach, it is not a field that has been subjected to much analysis as a whole. This paper provides an assessment of international education as a discipline, and outlines the global security framework. It examines how this framework is reflected in the forms of analysis used by international educationists. Finally it suggests how the central purpose of global security, namely ensuring human survival, could be adopted within international education to provide a clear sense of direction. This has specific implications for such areas as curriculum, assessment, educational provision and planning.
NASA Technical Reports Server (NTRS)
Hayes, J. D.
1972-01-01
The feasibility of monitoring volatile contaminants in a large space simulation chamber using techniques of internal reflection spectroscopy was demonstrated analytically and experimentally. The infrared spectral region was selected as the operational spectral range in order to provide unique identification of the contaminants along with sufficient sensitivity to detect trace contaminant concentrations. It was determined theoretically that a monolayer of the contaminants could be detected and identified using optimized experimental procedures. This ability was verified experimentally. Procedures were developed to correct the attenuated total reflectance spectra for thick sample distortion. However, by using two different element designs the need for such correction can be avoided.
NASA Astrophysics Data System (ADS)
Wang, Hong; Li, Xiufeng; Ge, Peng
2017-02-01
We propose a design method of an optical lens combined with a total internal reflection (TIR) freeform surface for a LED front fog lamp. The TIR freeform surface controls the edge rays of the LED source. It totally reflects the edge rays and makes them emit from the top surface of the lens. And the middle rays of the LED source go through the refractive surface and reach the measured plane. We simulate the model by Monte Carlo method. Simulation results show that the front fog lamp system can satisfy the requirement of ECE R19 Rev7. The light control efficiency can reach up to 76%.
78 FR 63433 - International Mailing Services: Proposed Price Changes-CPI
Federal Register 2010, 2011, 2012, 2013, 2014
2013-10-24
... POSTAL SERVICE 39 CFR Part 20 International Mailing Services: Proposed Price Changes--CPI AGENCY... mailing services price adjustments with the Postal Regulatory Commission (PRC), effective on January 26... Postal Service, International Mail Manual (IMM[supreg]) to reflect these new price changes. DATES: We...
78 FR 63434 - International Mailing Services: Proposed Price Changes-Exigent
Federal Register 2010, 2011, 2012, 2013, 2014
2013-10-24
... POSTAL SERVICE 39 CFR Part 20 International Mailing Services: Proposed Price Changes--Exigent... notice of mailing services price adjustments with the Postal Regulatory Commission (PRC), effective on... United States Postal Service, International Mail Manual (IMM[supreg]) to reflect these new price changes...
NASA Astrophysics Data System (ADS)
Tao, Xuemei; He, Yong
2006-09-01
The internal quality of tomato such as acidity and sugar content is important to its taste thus influences the market. The objective of this paper was to demonstrate the feasibility of using a near-infrared spectroscopy (NIRS) to investigate the relationship between sugar content and acidity of tomato and absorption spectra. The N1RS reflectance of nondestructive tomatoes was measured with a Visible/NJR spectrophotometer in 325-1075 nm range. The sugar content and acidity of tomato were obtained with a handhold sugar content meter and a PH meter. The reflectance data set was recorded and analyzed with some mathematic methods. The PLS (Partial least squares) calibration method was developed for converting the NIRS reflectance of tomato into the data which determined the acidity value. BP (Back propagation) neural network was used to set up the relationship between the NIRS reflectance of tomato and sugar content. The acidity values were detected with an accuracy of 9O% and the sugar contents determined by the BP network were also very close to the measurements (coefficient of correlation r2=0.8764). NW spectra analysis would be very useful in the nondestructive internal quality inspecting of tomato.
ERIC Educational Resources Information Center
Reis, Jehnie I.
2010-01-01
In the 1920s, French scholars and bureaucrats created the Cite Universitaire in Paris. The institution housed university students from around the world. The Cite founders formulated a model for the Cite that reflected ideological concerns in interwar Europe with a focus on pacifism, international education and cultural internationalism. The…
Executive reflects on progress in the oil and gas industry
DOE Office of Scientific and Technical Information (OSTI.GOV)
Popov, S.
1997-08-01
This paper reflects on the UK oil and gas industry`s international globalization and progress from the perspective of a UK industry executive. Sir Ian Wood, managing director of John Wood Group plc, outlined past and future industry developments during a 1997 Offshore Technology Conference speech. He concludes that the UK supply and service industry is now fully involved in the international arena, and hopes to play a significant role in the exciting oil and gas developments in the Gulf of Mexico and frontiers worldwide.
The Second Annual International Space University Alumni Conference
NASA Technical Reports Server (NTRS)
Johnson, Les (Compiler); Robinson, Paul A. (Compiler)
1994-01-01
The papers presented at the conference reflect the multidisciplinary nature of the International Space University (ISU) and its alumni. The first papers presented hold special relevance to the design projects, and cover such topics as lunar-based astronomical instrumentation, solar lunar power generation, habitation on the moon, and the legal issues governing multinational astronauts conducting research in space. The next set of papers cover various technical issues such as project success assessment, satellite networks and space station dynamics, thus reflecting the diverse backgrounds of the ISU alumni.
NASA Technical Reports Server (NTRS)
Mookherji, T.
1976-01-01
Outgassing, deposition, and desorption kinetics of silicone compounds, are examined as examples of optical surface contaminants of spacecraft windows. Their behavior in a space environment after exposure to ultraviolet radiation is also examined. The use of internal reflection spectroscopy is shown to provide a viable means of real-time, in-situ identification of contaminants of orbiting spacecraft. The instrumental techniques are proposed as the basis of further investigations and the development of flight hardware.
Hull, J R
1989-01-01
Coupling a dielectric compound parabolic concentrator (DCPC) to an absorber across a vacuum gap by means of frustrated total internal reflection (FTIR) can theoretically approach the maximum concentration permitted by physical laws, thus allowing higher radiative fluxes in thermal applications. The calculated optical performance of 2-D DCPCs with FTIR absorbers indicates that the ratio of gap thickness to optical wavelength must be <0.22 before the optical performance of the DCPC is superior to that of the nondielectric CPC.
Method of making a modular off-axis solar concentrator
DOE Office of Scientific and Technical Information (OSTI.GOV)
Plesniak, Adam P.; Hall, John C.
A method of making a solar concentrator may include forming a receiving wall having an elongated wall, a first side wall and a second side wall; attaching the first side wall and the second side wall to a reflecting wall to form a housing having an internal volume with an opening; forming a lip on the receiving wall and the reflecting wall; attaching a cover to the receiving wall and the reflecting wall at the lip to seal the opening into the internal volume, thereby creating a rigid structure; and mounting at least one receiver having at least one photovoltaicmore » cell on the elongated wall to receive solar radiation entering the housing and reflected by the receiving wall, the receiver having an axis parallel with a surface normal of the photovoltaic cell, such that the axis is disposed at a non-zero angle relative to the vertical axis of the opening.« less
Microwave based civil structure inspection device
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sohns, C.W.; Bible, D.W.
1994-06-01
A microwave based ``wall probe`` has been developed which is capable of nondestructive evaluation of architectural structures. By using microwaves in the 8 to 12 GHz range this probing instrument can detect subsurface characteristics through concrete, brick, wood or other building materials to depths in excess of 12 inches. The instrument interrogates a structure from a single side by transmitting a microwave signal into the surface at some angle of incidence and receiving the reflected signal some distance away on the same side of the structure. The transmitted signal is partially reflected at each internal boundary of different dielectric constant,more » giving a composite reflection which contains information from each internal layer. The reflected composite signal is compared in phase and amplitude to the transmitted signal and that reading is considered the ``signature`` of the structure under test. Computer algorithms analyze the signature for recognizable features and nonstandard construction.« less
Perspectives on Numeracy: Reflections from International Assessments
ERIC Educational Resources Information Center
Tout, Dave; Gal, Iddo
2015-01-01
This paper examines perspectives regarding the mathematical skills expected of adults and school graduates, comparing ideas developed as part of two major multinational comparative assessments of skills: the Programme for International Student Assessment and the Programme for International Assessment of Adult Competencies (also known as the OECD…
76 FR 76619 - International Mail: New Prices and Fee Changes-Mailing Services
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-08
... POSTAL SERVICE 39 CFR Part 20 International Mail: New Prices and Fee Changes--Mailing Services... Individual Country Listings (ICLs) to reflect price adjustments for First-Class Mail International[supreg... mailing services price adjustments with the Postal Regulatory Commission (PRC), effective on January 22...
75 FR 5351 - Proposed Revisions to Accounting Guide for LSC Recipients
Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-02
... elements of an adequate accounting and financial reporting system, including the use of specific internal... checklist of accounting procedures and internal controls. The proposed revisions update the checklist to... accounting procedures and internal controls to reflect current best practices; (7) updated and new references...
Factor Analysis of Intern Effectiveness
ERIC Educational Resources Information Center
Womack, Sid T.; Hannah, Shellie Louise; Bell, Columbus David
2012-01-01
Four factors in teaching intern effectiveness, as measured by a Praxis III-similar instrument, were found among observational data of teaching interns during the 2010 spring semester. Those factors were lesson planning, teacher/student reflection, fairness & safe environment, and professionalism/efficacy. This factor analysis was as much of a…
Driver, Christine
2005-04-01
This paper explores the dynamics brought into analytic work when there is a symmetric fusion between psyche and soma within the patient. It will consider how such a fusion may emerge from reverberations between physical constitution and a lack of maternal attunement, containment and reflective function. I will describe the work with a patient, Jane, who was diagnosed with Myalgic Encephalomyelitis (ME) during the course of her analysis. The dynamic of her physical symptoms within the analytic work, and the impact of her internal affects and internal 'objects' within the transference and countertransference, indicated a difficulty in finding an homeostatic balance resulting in overactivity and underactivity at both somatic and psychological levels. Using the clinical work with Jane this paper will also examine the interrelationship between mother-infant attachment, an inadequate internalized maternal reflective function, affect dysregulation, unconscious fusion, the lack of psyche-soma differentiation and the impact of the latter in relation to internal regulation systems, or lack of, in patients with Chronic Fatigue Syndrome (CFS) and Myalgic Encephalomyelitis (ME). I will draw on similar work carried out by Holland (1997), Simpson (1997) and Simpson et al. (1997). The paper will also employ the concept of the reflective function (Fonagy 2001; Knox 2003), and consider Matte-Blanco's (1999) concepts of generalization and unconscious symmetry in relation to the patient's internal world. I go on to consider how analysis provides a point outside the 'fusion' that can enable the 'deadlock' to be broken.
Near-surface energy transfers from internal tide beams to smaller vertical scale motions
NASA Astrophysics Data System (ADS)
Chou, S.; Staquet, C.; Carter, G. S.; Luther, D. S.
2016-02-01
Mechanical energy capable of causing diapycnal mixing in the ocean is transferred to the internal wave field when barotropic tides pass over underwater topography and generate internal tides. The resulting internal tide energy is confined in vertically limited structures, or beams. As internal tide beams (ITBs) propagate through regions of non-uniform stratification in the upper ocean, wave energy can be scattered through multiple reflections and refractions, be vertically trapped, or transferred to non-tidal frequencies through different nonlinear processes. Various observations have shown that ITBs are no longer detectable in horizontal kinetic energy beyond the first surface reflection. Importantly, this implies that some of the internal tide energy no longer propagates in to the abyssal ocean and consequently will not be available to maintain the density stratification. Using the NHM, a nonlinear and nonhydrostatic model based on the MITgcm, simulations of an ITB propagating up to the sea surface are examined in order to quantify the transformation of ITB energy to other motions. We compare and contrast the transformations enabled by idealized, smoothly-varying stratification with transformations enabled by realistic stratification containing a broad-band vertical wavenumber spectrum of variations. Preliminary two-dimensional results show that scattering due to small-scale structure in realistic stratification profiles from Hawaii can lead to energy being vertically trapped near the surface. Idealized simulations of "locally" generated internal solitary waves are analyzed in terms of energy flux transfers from the ITB to solitary waves, higher harmonics, and mean flow. The amount of internal tide energy which propagates back down after near-surface reflection of the ITB in different environments is quantified.
ELiXIR—Solid-State Luminaire With Enhanced Light Extraction by Internal Reflection
NASA Astrophysics Data System (ADS)
Allen, Steven C.; Steckl, Andrew J.
2007-06-01
A phosphor-converted light-emitting diode (pcLED) luminaire featuring enhanced light extraction by internal reflection (ELiXIR) with efficacy of 60 lm/W producing 18 lumens of yellowish green light at 100 mA is presented. The luminaire consists of a commercial blue high power LED, a polymer hemispherical shell lens with interior phosphor coating, and planar aluminized reflector. High extraction efficiency of the phosphor-converted light is achieved by separating the phosphor from the LED and using internal reflection to steer the light away from lossy reflectors and the LED package and out of the device. At 10 and 500 mA, the luminaire produces 2.1 and 66 lumens with efficacies of 80 and 37 lm/W, respectively. Technological improvements over existing commercial LEDs, such as more efficient pcLED packages or, alternatively, higher efficiency green or yellow for color mixing, will be essential to achieving 150 200 lm/W solid-state lighting. Advances in both areas are demonstrated.
Factors confounding the assessment of reflection: a critical review
2011-01-01
Background Reflection on experience is an increasingly critical part of professional development and lifelong learning. There is, however, continuing uncertainty about how best to put principle into practice, particularly as regards assessment. This article explores those uncertainties in order to find practical ways of assessing reflection. Discussion We critically review four problems: 1. Inconsistent definitions of reflection; 2. Lack of standards to determine (in)adequate reflection; 3. Factors that complicate assessment; 4. Internal and external contextual factors affecting the assessment of reflection. Summary To address the problem of inconsistency, we identified processes that were common to a number of widely quoted theories and synthesised a model, which yielded six indicators that could be used in assessment instruments. We arrived at the conclusion that, until further progress has been made in defining standards, assessment must depend on developing and communicating local consensus between stakeholders (students, practitioners, teachers, supervisors, curriculum developers) about what is expected in exercises and formal tests. Major factors that complicate assessment are the subjective nature of reflection's content and the dependency on descriptions by persons being assessed about their reflection process, without any objective means of verification. To counter these validity threats, we suggest that assessment should focus on generic process skills rather than the subjective content of reflection and where possible to consider objective information about the triggering situation to verify described reflections. Finally, internal and external contextual factors such as motivation, instruction, character of assessment (formative or summative) and the ability of individual learning environments to stimulate reflection should be considered. PMID:22204704
Building an environmentally accountable medical curriculum through international collaboration.
Walpole, Sarah Catherine; Vyas, Aditya; Maxwell, Janie; Canny, Ben J; Woollard, Robert; Wellbery, Caroline; Leedham-Green, Kathleen E; Musaeus, Peter; Tufail-Hanif, Uzma; Pavão Patrício, Karina; Rother, Hanna-Andrea
2017-10-01
Global environmental change is associated with significant health threats. The medical profession can address this challenge through advocacy, health system adaptation and workforce preparedness. Stewardship of health systems with attention to their environmental impacts can contribute to mitigation of and adaptation to negative health impacts of environmental change. Medical schools have an integral role in training doctors who understand the interdependence of ecosystems and human health. Yet integrating environmental perspectives into busy medical curricula is not a simple task. At the 2016 Association for Medical Education in Europe conference, medical educators, students and clinicians from six continents discussed these challenges in a participatory workshop. Here we reflect on emerging themes from the workshop and how to plan for curricular change. Firstly, we outline recent developments in environmental health and associated medical education. Secondly, we reflect on our process and outcomes during this innovative approach to international collaboration. Thirdly, we present learning objectives which cover core content for environmentally accountable medical curricula, developed through a reflective process of international collaboration integrating current literature and the workshop outcomes. International collaboration can bring together diverse perspectives and provide critical insights for the inclusion of environmental health into basic education for medical practitioners.
26 CFR 1.897-2 - United States real property holding corporations.
Code of Federal Regulations, 2010 CFR
2010-04-01
.... (iii) Denial of presumption. If the Internal Revenue Service determines, on the basis of information as.... 1.897-2 Section 1.897-2 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... (b)(2) may not accurately reflect the status of the corporation, the Service will notify the...
ERIC Educational Resources Information Center
Burkholder, Jessica Reno
2014-01-01
A modified version of Moustakas' (1994) method of analyzing phenomenological data was used to illuminate how full-time, single, Turkish international graduate students conceptualized their experiences as international students. The participants detailed common and salient aspects of their experience: personal growth, decisions regarding…
Living and Learning Internationally for a Viable Future: Reflecting on UNESCO CONFINTEA VI
ERIC Educational Resources Information Center
Henschke, John A.
2012-01-01
International travel has the potential to expand one's learning horizons exponentially, especially with the abundantly enriching experiences of adult, continuing, and lifelong learning conferences. CONFINTEA VI (The Sixth World International Conference in Adult Education) sponsored by the United Nations Educational, Scientific, and Cultural…
Troubling Metaphors and International Student Adjustment: Reflections from a Transnational Place
ERIC Educational Resources Information Center
Starr-Glass, David
2017-01-01
On many campuses, offices of International Student Affairs address the perceived needs of international students. However, a number of underlying assumptions and persistent metaphors shape these efforts and influence their outcomes. All students are uniquely different and face equally different challenges in adjusting to higher education. Labeling…
ERIC Educational Resources Information Center
Thammasat Univ., Bangkok (Thailand). Univ. Libraries.
This document which covers the proceedings of the 1989 International Symposium on Information Technology, begins with several opening ceremony messages and includes the following papers: (1) "Reflections on International Bibliographic Standards" (Winston D. Roberts); (2) "Bibliographic Control from the User's Perspective" (Ruth…
Reflections on a Seminal Force in International Accounting
ERIC Educational Resources Information Center
Cascini, Karen T.
2007-01-01
Accounting is a manifestation of several important environmental factors within a country, including economic, educational and political, and, as such, is evolutionary in accordance with those changing social structures. Because of the major impact that international accounting has had on countries' internal accounting systems, it is important to…
The Experience of an International University Teacher in Australia
ERIC Educational Resources Information Center
Abayadeera, Nadana
2013-01-01
Accounting and Business schools in Australia have a considerable number of international academic staff, and the teaching effectiveness of such staff is an increasingly salient issue. Many international teachers are non-native English speakers and have different teaching approaches and styles, often reflecting their own experiences as a learner in…
Systems and Methods for Correcting Optical Reflectance Measurements
NASA Technical Reports Server (NTRS)
Yang, Ye (Inventor); Shear, Michael A. (Inventor); Soller, Babs R. (Inventor); Soyemi, Olusola O. (Inventor)
2014-01-01
We disclose measurement systems and methods for measuring analytes in target regions of samples that also include features overlying the target regions. The systems include: (a) a light source; (b) a detection system; (c) a set of at least first, second, and third light ports which transmit light from the light source to a sample and receive and direct light reflected from the sample to the detection system, generating a first set of data including information corresponding to both an internal target within the sample and features overlying the internal target, and a second set of data including information corresponding to features overlying the internal target; and (d) a processor configured to remove information characteristic of the overlying features from the first set of data using the first and second sets of data to produce corrected information representing the internal target.
Systems and methods for correcting optical reflectance measurements
NASA Technical Reports Server (NTRS)
Yang, Ye (Inventor); Soller, Babs R. (Inventor); Soyemi, Olusola O. (Inventor); Shear, Michael A. (Inventor)
2009-01-01
We disclose measurement systems and methods for measuring analytes in target regions of samples that also include features overlying the target regions. The systems include: (a) a light source; (b) a detection system; (c) a set of at least first, second, and third light ports which transmit light from the light source to a sample and receive and direct light reflected from the sample to the detection system, generating a first set of data including information corresponding to both an internal target within the sample and features overlying the internal target, and a second set of data including information corresponding to features overlying the internal target; and (d) a processor configured to remove information characteristic of the overlying features from the first set of data using the first and second sets of data to produce corrected information representing the internal target.
Theoretical analysis of non-polarizing beam splitters with appropriate amplitude and phase
NASA Astrophysics Data System (ADS)
Shi, Jin Hui; Wang, Zheng Ping; Guan, Chun Ying
2009-04-01
When used at oblique angles of incidence, the reflectance and transmittance of thin films exhibit strong polarization effects, particularly for the films inside a glass cube. However, the polarization effects are undesirable in many applications. Novel non-polarizing beam splitter designs are shown. Non-polarizing beam splitters with unique optical thin films are achieved through the combination of interference and frustrated total internal reflection. The non-polarizing condition expressions based on frustrated total internal reflection are derived, and examples of the non-polarizing beam splitters are also presented with the optimization technique and the results of Rp=(50±0.5)%, Rs=(50±0.5)%, and Δr=(0±0.3) in the wavelength range of 400-700 nm are obtained.
ERIC Educational Resources Information Center
Fierke, Kerry K.; Lepp, Gardner A.; Bastianelli, Karen; Vogelsang, Lisa; Tornabene, Ladona
2016-01-01
The article describes a student-centered approach to generating meaningful learning outcomes in a short-term study abroad program. A practice named Intention/Reflection (I/R) was used to help students to identify, articulate, and reflect upon learning objectives that were personally meaningful, within the broader framework of the intended outcomes…
Increased efficiency with surface texturing in ITO/InP solar cells
NASA Technical Reports Server (NTRS)
Jenkins, Phillip; Landis, Geoffrey A.; Fatemi, Navid; Li, Xiaonan; Scheiman, David; Bailey, Sheila
1992-01-01
Optimization of an InP solar cell with a V-grooved surface is discussed. Total internal reflection in the coverglass reduces surface reflection and can recover light reflected from the front metallization. Results from the first ITO/InP solar cells on low-angle V-grooved substrates are presented, showing a 5.8 percent increase in current.
ERIC Educational Resources Information Center
Pfeifer, Jennifer H.; Masten, Carrie L.; Borofsky, Larissa A.; Dapretto, Mirella; Fuligni, Andrew J.; Lieberman, Matthew D.
2009-01-01
Classic theories of self-development suggest people define themselves in part through internalized perceptions of other people's beliefs about them, known as reflected self-appraisals. This study uses functional magnetic resonance imaging to compare the neural correlates of direct and reflected self-appraisals in adolescence (N = 12, ages 11-14…
Code of Federal Regulations, 2011 CFR
2011-04-01
... 26 Internal Revenue 4 2011-04-01 2011-04-01 false Adjustments to earnings and profits reflecting... § 1.312-9 Adjustments to earnings and profits reflecting increase in value accrued before March 1..., that part of the earnings and profits which is represented by increase in value of property accrued...
ERIC Educational Resources Information Center
Riedler, Martina
2016-01-01
In this self-reflective auto-ethnographic research, the author shares her experiences of introspection, change and professional growth as an art educator in an international context. Auto-ethnography is an approach to qualitative inquiry in which the researcher employs self-reflection to explore her personal experiences and connect these…
Code of Federal Regulations, 2010 CFR
2010-04-01
... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Disclosures of return information reflected on returns to officers and employees of the Department of Commerce for certain statistical purposes and... reflected on returns to officers and employees of the Department of Commerce for certain statistical...
Fourier Transform Infrared Spectroscopy Part III. Applications.
ERIC Educational Resources Information Center
Perkins, W. D.
1987-01-01
Discusses the use of the FT-IR spectrometer in analyses that were previously avoided. Examines some of the applications of this spectroscopy with aqueous solutions, circular internal reflection, samples with low transmission, diffuse reflectance, infrared emission, and the infrared microscope. (TW)
Reflections on the Medical Library Association's international activities.
Poland, U H
1982-01-01
An overview of the Medical Library Association's past international activities is given with emphasis on the international fellowship program, international exchange of materials, participation in the International Federation of Library Associations, and international congresses on medical librarianship. Problems presented by cultural and educational differences, as well as governmental, political, and economic influences affecting international activities are enumerated. Lastly, continuation of the association's current international activities is endorsed, especially the extension of bilateral agreements with health sciences library associations of other countries, and increased activity in comparative medical librarianship. PMID:7150824
Fate of internal waves on a shallow shelf
NASA Astrophysics Data System (ADS)
Davis, Kristen; Arthur, Robert; Reid, Emma; Decarlo, Thomas; Cohen, Anne
2017-11-01
Internal waves strongly influence the physical and chemical environment of coastal ecosystems worldwide. We report novel observations from a distributed temperature sensing (DTS) system that tracked the transformation of internal waves from the shelf break to the surf zone over a shelf-slope region of a coral atoll in the South China Sea. The spatially-continuous view of the near-bottom temperature field provided by the DTS offers a perspective of physical processes previously available only in laboratory settings or numerical models. These processes include internal wave reflection off a natural slope, shoreward transport of dense fluid within trapped cores, internal ``tide pools'' (dense water left behind after the retreat of an internal wave), and internal run-down (near-bottom, offshore-directed jets of water preceding a breaking internal wave). Analysis shows that the fate of internal waves on this shelf - whether they are transmitted into shallow waters or reflected back offshore - is mediated by local water column density and shear structure, with important implications for nearshore distributions of energy, heat, and nutrients. We acknowledge the US Army Research Laboratory DoD Supercomputing Resource Center for computer time on Excalibur, which was used for the numerical simulations in this work. Funding for field work supported by Academia Sinica and for K.D. and E.R. from NSF.
Wang, Shinn-Fwu
2009-01-01
A small-displacement sensor based on total-internal reflection theory and surface plasmon resonance technology is proposed for use in heterodyne interferometry. A small displacement can be obtained simply by measuring the variation in phase difference between s- and p-polarization states with the small-displacement sensor. The theoretical displacement resolution of the small-displacement sensor can reach 0.45 nm. The sensor has some additional advantages, e.g., a simple optical setup, high resolution, high sensitivity and rapid measurement. Its feasibility is also demonstrated.
NASA Astrophysics Data System (ADS)
Nabok, A. V.; Tsargorodskaya, A.; Hassan, A. K.; Starodub, N. F.
2005-06-01
The environmental toxins, such as herbicides simazine and atrazine, and T2 mycotoxin were registered with the optical methods of surface plasmon resonance (SPR) and recently developed total internal reflection ellipsometry (TIRE). The immune assay approach was exploited for in situ registration of the above low molecular weight toxins with specific antibodies immobilised onto the gold surface via (poly)allylamine hydrochloride layer using electrostatic self-assembly (ESA) technique. The comparison of two methods of SPR and TIRE shows a higher sensitivity of the latter.
Wallhead, Ian; Jiménez, Teresa Molina; Ortiz, Jose Vicente García; Toledo, Ignacio Gonzalez; Toledo, Cristóbal Gonzalez
2012-11-05
A novel of Fresnel-type lens for use as a solar collector has been designed which utilizes double total internal reflection (D-TIR) to optimize collection efficiency for high numerical aperture lenses (in the region of 0.3 to 0.6 NA). Results show that, depending on the numerical aperture and the size of the receiver, a collection efficiency theoretical improvement on the order of 20% can be expected with this new design compared with that of a conventional Fresnel lens.
Sawatsky, Adam P; Nordhues, Hannah C; Merry, Stephen P; Bashir, M Usmaan; Hafferty, Frederic W
2018-03-27
International health electives (IHEs) are widely available during residency and provide unique experiences for trainees. Theoretical models of professional identity formation and transformative learning may provide insight into residents' experiences during IHEs. The purpose of this study was to explore transformative learning and professional identity formation during resident IHEs and characterize the relationship between transformative learning and professional identity formation. The authors used a constructivist grounded theory approach, with the sensitizing concepts of transformative learning and professional identity formation to analyze narrative reflective reports of residents' IHEs. The Mayo International Health Program supports residents from all specialties across three Mayo Clinic sites. In 2015, the authors collected narrative reflective reports from 377 IHE participants dating from 2001-2014. Reflections were coded and themes were organized into a model for transformative learning during IHEs, focusing on professional identity. Five components of transformative learning were identified during IHEs: a disorienting experience; an emotional response; critical reflection; perspective change; and a commitment to future action. Within the component of critical reflection three domains relating to professional identity were identified: making a difference; the doctor-patient relationship; and medicine in its "purest form." Transformation was demonstrated through perspective change and a commitment to future action, including continued service, education, and development. IHEs provide rich experiences for transformative learning and professional identity formation. Understanding the components of transformative learning may provide insight into the interaction between learner, experiences, and the influence of mentors in the process of professional identity formation.
Internality and Achievement in the Intermediate Grades.
ERIC Educational Resources Information Center
Creek, Roy J.; And Others
Locus of control is a construct that reflects an individual's perception of control over his or her own destiny. The thesis is that people adopt either an internal or an external orientation. Internally oriented persons consider success the result of ability and effort. Externally oriented individuals attribute success to luck, fate, or powerful…
Federal Register 2010, 2011, 2012, 2013, 2014
2013-05-29
... International Student Assessment (PISA 2015) Recruitment and Field Test AGENCY: Department of Education. ACTION..., page 22530) for the Program for International Student Assessments (PISA 2015) Recruitment and Field Test, 1850-0755. Since that time, the PISA has been amended. It reflects a change in the field test...
ERIC Educational Resources Information Center
Streets, Barbara Faye; Nicolas, Guerda; Wolford, Karen
2015-01-01
International service learning courses, cultural immersion projects, and international disaster response teams have provided valuable aid, services, supplies and programs to trauma-impacted communities across the globe. Many colleges and universities support global learning and the creation of global citizens, and this ethic is reflected in many…
Social Network Analysis of International Student Mobility: Uncovering the Rise of Regional Hubs
ERIC Educational Resources Information Center
Kondakci, Yasar; Bedenlier, Svenja; Zawacki-Richter, Olaf
2018-01-01
Research on the patterns of international student mobility and the dynamics shaping these patterns has been dominated by studies reflecting a Western orientation, discourse, and understanding. Considering political, economic, cultural, historical, and ecological factors, this study argues that international student mobility is not only an issue of…
ERIC Educational Resources Information Center
Parks, Elizabeth
2015-01-01
Linguistic ideologies that are left unquestioned and unexplored, especially as reflected and produced in marginalized language communities, can contribute to inequality made real in decisions about languages and the people who use them. One of the primary bodies of knowledge guiding international language policy is the International Organization…
ERIC Educational Resources Information Center
Burkholder, Jessica Reno
2010-01-01
The research was guided by the research question: How do full-time single Turkish international graduate students conceptualize their experiences as international students? Participants in the study included three doctoral students and three master's students who participated in a series of semi-structured interviews. The data was transcribed and…
International Encounters: Experiences of Visitors and Hosts in a Cross-Cultural Program
ERIC Educational Resources Information Center
Ranz, Rebecca; Nuttman-Shwartz, Orit; Thachil, Githa
2015-01-01
The present study aimed to gain further insights into how visiting international social work students and their hosts from different cultures learn about each others' social services, culture, and personal values. Six Israeli students and 8 Indian students have written narratives and reflections on their experiences in the international encounter…
Science and Mathematics Education: International Innovations, Research, and Practices
ERIC Educational Resources Information Center
Berlin, Donna F., Ed.; White, Arthur L., Ed.
2011-01-01
The chapters in this book reflect the work of science and mathematics educators who have worked for many years at the international level. As members of the International Consortium for Research in Science and Mathematics Education, their work provides readers with issues, models, practices, and research results that have applicability and…
Ellefsen, Kyle L; Settle, Brett; Parker, Ian; Smith, Ian F
2014-09-01
Local Ca(2+) transients such as puffs and sparks form the building blocks of cellular Ca(2+) signaling in numerous cell types. They have traditionally been studied by linescan confocal microscopy, but advances in TIRF microscopy together with improved electron-multiplied CCD (EMCCD) cameras now enable rapid (>500 frames s(-1)) imaging of subcellular Ca(2+) signals with high spatial resolution in two dimensions. This approach yields vastly more information (ca. 1 Gb min(-1)) than linescan imaging, rendering visual identification and analysis of local events imaged both laborious and subject to user bias. Here we describe a routine to rapidly automate identification and analysis of local Ca(2+) events. This features an intuitive graphical user-interfaces and runs under Matlab and the open-source Python software. The underlying algorithm features spatial and temporal noise filtering to reliably detect even small events in the presence of noisy and fluctuating baselines; localizes sites of Ca(2+) release with sub-pixel resolution; facilitates user review and editing of data; and outputs time-sequences of fluorescence ratio signals for identified event sites along with Excel-compatible tables listing amplitudes and kinetics of events. Copyright © 2014 Elsevier Ltd. All rights reserved.
Imaging of DNA and Protein by SFM and Combined SFM-TIRF Microscopy.
Grosbart, Małgorzata; Ristić, Dejan; Sánchez, Humberto; Wyman, Claire
2018-01-01
Direct imaging is invaluable for understanding the mechanism of complex genome transactions where proteins work together to organize, transcribe, replicate and repair DNA. Scanning (or atomic) force microscopy is an ideal tool for this, providing 3D information on molecular structure at nm resolution from defined components. This is a convenient and practical addition to in vitro studies as readily obtainable amounts of purified proteins and DNA are required. The images reveal structural details on the size and location of DNA bound proteins as well as protein-induced arrangement of the DNA, which are directly correlated in the same complexes. In addition, even from static images, the different forms observed and their relative distributions can be used to deduce the variety and stability of different complexes that are necessarily involved in dynamic processes. Recently available instruments that combine fluorescence with topographic imaging allow the identification of specific molecular components in complex assemblies, which broadens the applications and increases the information obtained from direct imaging of molecular complexes. We describe here basic methods for preparing samples of proteins, DNA and complexes of the two for topographic imaging and quantitative analysis. We also describe special considerations for combined fluorescence and topographic imaging of molecular complexes.
Resident CAPS on dense-core vesicles docks and primes vesicles for fusion
Kabachinski, Greg; Kielar-Grevstad, D. Michelle; Zhang, Xingmin; James, Declan J.; Martin, Thomas F. J.
2016-01-01
The Ca2+-dependent exocytosis of dense-core vesicles in neuroendocrine cells requires a priming step during which SNARE protein complexes assemble. CAPS (aka CADPS) is one of several factors required for vesicle priming; however, the localization and dynamics of CAPS at sites of exocytosis in live neuroendocrine cells has not been determined. We imaged CAPS before, during, and after single-vesicle fusion events in PC12 cells by TIRF microscopy. In addition to being a resident on cytoplasmic dense-core vesicles, CAPS was present in clusters of approximately nine molecules near the plasma membrane that corresponded to docked/tethered vesicles. CAPS accompanied vesicles to the plasma membrane and was present at all vesicle exocytic events. The knockdown of CAPS by shRNA eliminated the VAMP-2–dependent docking and evoked exocytosis of fusion-competent vesicles. A CAPS(ΔC135) protein that does not localize to vesicles failed to rescue vesicle docking and evoked exocytosis in CAPS-depleted cells, showing that CAPS residence on vesicles is essential. Our results indicate that dense-core vesicles carry CAPS to sites of exocytosis, where CAPS promotes vesicle docking and fusion competence, probably by initiating SNARE complex assembly. PMID:26700319
Seasonal transitions and social responsibility.
Small, Mary Rose
2013-01-01
This paper is a reflection on work transitions in an international context. Seasonal employees and employer of the winery Emiliana. The following is a personal reflection by a Canadian occupational therapy student after a recent trip to Chile. The commitment to social responsibility by the winery Emiliana guided the reflection on work transitions of seasonal employees and the employer's provision of services to improve employee's quality of life.
ERIC Educational Resources Information Center
Baker, Fiona S.
2015-01-01
This article discusses a teacher educator's reflections on her participation in an international study group and visits to the infant-toddler and pre-schools of Reggio Emilia, Italy, as inspiration for early childhood teacher education in Abu Dhabi, UAE. The following five themes are reflected on, for teacher education in the context of Abu Dhabi:…
Opportunities to Intercalibrate Radiometric Sensors From International Space Station
NASA Technical Reports Server (NTRS)
Roithmayr, C. M.; Lukashin, C.; Speth, P. W.; Thome, K. J.; Young, D. F.; Wielicki, B. A.
2012-01-01
Highly accurate measurements of Earth's thermal infrared and reflected solar radiation are required for detecting and predicting long-term climate change. We consider the concept of using the International Space Station to test instruments and techniques that would eventually be used on a dedicated mission such as the Climate Absolute Radiance and Refractivity Observatory. In particular, a quantitative investigation is performed to determine whether it is possible to use measurements obtained with a highly accurate reflected solar radiation spectrometer to calibrate similar, less accurate instruments in other low Earth orbits. Estimates of numbers of samples useful for intercalibration are made with the aid of year-long simulations of orbital motion. We conclude that the International Space Station orbit is ideally suited for the purpose of intercalibration.
Diary of a (new) parliamentary intern.
Ridge, Anthony
2015-10-17
Anthony Ridge is the new parliamentary intern to Lord Trees. Here, he reflects on his new job and discovering his place in the veterinary profession. Anthony will be sharing his experiences with Vet Record Careers. British Veterinary Association.
[Concept analysis of reflective thinking].
Van Vuuren, M; Botes, A
1999-09-01
The nursing practice is described as a scientific practice, but also as a practice where caring is important. The purpose of nursing education is to provide competent nursing practitioners. This implies that future practitioners must have both critical analytical thinking abilities, as well as empathy and moral values. Reflective thinking could probably accommodate these thinking skills. It seems that the facilitation of reflective thinking skills is essential in nursing education. The research question that is relevant in this context is: "What is reflective thinking?" The purpose of this article is to report on the concept analysis of reflective thinking and in particular on the connotative meaning (critical attributes) thereof. The method used to perform the concept analysis is based on the original method of Wilson (1987) as described by Walker & Avant (1995). As part of the concept analysis the connotations (critical attributes) are identified, reduced and organized into three categories, namely pre-requisites, processes and outcomes. A model case is described which confirms the essential critical attributes of reflective thinking. Finally a theoretical definition of reflective thinking is derived and reads as follows: Reflective thinking is a cyclic, hierarchical and interactive construction process. It is initiated, extended and continued because of personal cognitive-affective interaction (individual dimension) as well as interaction with the social environment (social dimension). to realize reflective thinking, a level of internalization on the cognitive and affective domain is required. The result of reflective thinking is a integrated framework of knowledge (meaningful learning) and a internalized value system providing a new perspective on and better understanding of a problem. Reflective thinking further leads to more effective decision making- and problem solving skills.
Kuang, Cuifang; Ali, M Yakut; Hao, Xiang; Wang, Tingting; Liu, Xu
2010-10-01
In order to achieve a higher axial resolution for displacement measurement, a novel method is proposed based on total internal reflection filter and confocal microscope principle. A theoretical analysis of the basic measurement principles is presented. The analysis reveals that the proposed confocal detection scheme is effective in enhancing the resolution of nonlinearity of the reflectance curve greatly. In addition, a simple prototype system has been developed based on the theoretical analysis and a series of experiments have been performed under laboratory conditions to verify the system feasibility, accuracy, and stability. The experimental results demonstrate that the axial resolution in displacement measurements is better than 1 nm in a range of 200 nm which is threefold better than that can be achieved using the plane reflector.
Design and analysis of non-polarizing beam splitter in a glass cube
NASA Astrophysics Data System (ADS)
Shi, Jinhui; Wang, Zhengping; Huang, Zongjun; Li, Qingbo
2008-11-01
The reflectance and transmittance of thin films at oblique incident angles exhibit strong polarization effects, particularly for the films inside a glass cube. However, the polarization effects are undesirable in many applications. To solve this problem, non-polarizing beam splitters with unique optical thin films have been achieved employing a method of combination of interference and frustrated total internal reflection, the non-polarizing condition expressions based on frustrated total internal reflection has been derived, and the design examples of non-polarizing beam splitters with an optimization technique have been also presented. The results of Rp=(50+/-0.5)%, Rs=(50+/-0.5)% andΔr=(0+/-0.3) degree in the wavelength range of 400-700nm have been obtained. The thickness sensitivity of NPBSs is also analyzed.
Internal reflection sensors with high angular resolution
NASA Astrophysics Data System (ADS)
Shavirin, I.; Strelkov, O.; Vetskous, A.; Norton-Wayne, L.; Harwood, R.
1996-07-01
We discuss the use of total internal reflection for the production of sensors with high angular resolution. These sensors are intended for measurement of the angle between a sensor's axis and the direction to a source of radiation or reflecting object. Sensors of this type are used in controlling the position of machine parts in robotics and industry, orienting space vehicles and astronomic devices in relation to the Sun, and as autocollimators for checking angles of deviation. This kind of sensor was used in the Apollo space vehicle some 20 years ago. Using photodetectors with linear and area CCD arrays has opened up new application possibilities for appropriately designed sensors. A generalized methodology is presented applicable to a wide range of tasks. Some modifications that can improve the performance of the basic design are described.
Energy loss from internal reflection off metal layers on glass
NASA Astrophysics Data System (ADS)
McDowell, M. W.; Bezuidenhout, D. F.; Klee, H. W.; Theron, E.
1983-12-01
The reflection characteristics of metal layers are considered for the situation where the electromagnetic radiation is incident from the glass side. Theoretical and measured reflectance values are presented which indicate that for some metals the reflection has a strong dependence on the refractive index of the incident medium. Some examples are given of recent cases where the above results were an important consideration in the choice of the metallic reflecting material. These results indicate that aluminium should not be automatically considered the best choice for the visible region nor gold for the infra-red.
Reflections on CME Congress 2012
ERIC Educational Resources Information Center
Knox, Alan B.
2013-01-01
This commentary reflects the author's impressions of Continuing Medical Education (CME) Congress 2012, a provocative international conference on professional development and quality improvement in the health professions that took place in Toronto, Ontario, last spring. The sessions he attended and conversations he had with other attendees were…
Reflectance and internal structure of leaves from several crops during a growing season.
NASA Technical Reports Server (NTRS)
Sinclair, T. R.; Hoffer, R. M.; Schreiber, M. M.
1971-01-01
Measurements of spectral reflectance characteristics during a growing season of leaves from six crops are reported. These crops include soybeans, wheat, oats, sorghum, corn, and sudangrass. The characteristics measured are related to changes in leaf structure and water content.
Schooling Redesigned: Towards Innovative Learning Systems
ERIC Educational Resources Information Center
Istance, David
2015-01-01
What does redesigning schools and schooling through innovation mean in practice? How might it be brought about? These questions have inspired an influential international reflection on "Innovative Learning Environments" (ILE) led by the OECD. This reflection has already resulted in publications on core design principles and frameworks…
Using a laser source to measure the refractive index of glass beads and Debye theory analysis.
Li, Shui-Yan; Qin, Shuang; Li, Da-Hai; Wang, Qiong-Hua
2015-11-20
Using a monochromatic laser beam to illuminate a homogeneous glass bead, some rainbows will appear around it. This paper concentrates on the study of the scattering intensity distribution and the method of measuring the refractive index for glass beads based on the Debye theory. It is found that the first rainbow due to the scattering superposition of backward light of the low-refractive-index glass beads can be explained approximately with the diffraction, the external reflection plus the one internal reflection, while the second rainbow of high-refractive-index glass beads is due to the contribution from the diffraction, the external reflection, the direct transmission, and the two internal reflections. The scattering intensity distribution is affected by the refractive index, the radius of the glass bead, and the incident beam width. The effects of the refractive index and the glass bead size on the first and second minimum deviation angle position are analyzed in this paper. The results of the measurements agree very well with the specifications.
Resident Self-Assessment and Self-Reflection: University of Wisconsin-Madison’s Five-Year Study
Hildebrand, Christopher; Trowbridge, Elizabeth; Roach, Mary A.; Sullivan, Anne Gravel; Broman, Aimee Teo
2009-01-01
BACKGROUND Chart review represents a critical cornerstone for practice-based learning and improvement in our internal medicine residency program. OBJECTIVE To document residents’ performance monitoring and improvement skills in their continuity clinics, their satisfaction with practice-based learning and improvement, and their ability to self-reflect on their performance. DESIGN Retrospective longitudinal design with repeated measures. PARTICIPANTS Eighty Internal Medicine residents abstracted data for 3 consecutive years from the medical records of their 4,390 patients in the University of Wisconsin-Madison (UW) Hospital and Clinics and William S. Middleton Veterans Administration (VA) outpatient clinics. MEASUREMENT Logistic modeling was used to determine the effect of postgraduate year, resident sex, graduation cohort, and clinic setting on residents’ “compliance rate” on 17 nationally recognized health screening and chronic disease management parameters from 2003 to 2007. RESULTS Residents’ adherence to national preventive and chronic disease standards increased significantly from intern to subsequent years for administering immunizations, screening for diabetes, cholesterol, cancer, and behavioral risks, and for management of diabetes. Of the residents, 92% found the chart review exercise beneficial, with 63% reporting gains in understanding about their medical practices, 26% reflecting on specific gaps in their practices, and 8% taking critical action to improve their patient outcomes. CONCLUSIONS This paper provides support for the feasibility and practicality of this limited-cost method of chart review. It also directs our residency program’s attention in the continuity clinic to a key area important to internal medicine training programs by highlighting the potential benefit of enhancing residents’ self-reflection skills. PMID:19156469
The utility of vignettes to stimulate reflection on professionalism: theory and practice.
Bernabeo, E C; Holmboe, E S; Ross, K; Chesluk, B; Ginsburg, S
2013-08-01
Professionalism remains a substantive theme in medical literature. There is an emerging emphasis on sociological and complex adaptive systems perspectives that refocuses attention from just the individual role to working within one's system to enact professionalism in practice. Reflecting on responses to professional dilemmas may be one method to help practicing physicians identify both internal and external factors contributing to (un) professional behavior. We present a rationale and theoretical framework that supports and guides a reflective approach to the self assessment of professionalism. Guided by principles grounded in this theoretical framework, we developed and piloted a set of vignettes on professionally challenging situations, designed to stimulate reflection in practicing physicians. Findings show that participants found the vignettes to be authentic and typical, and reported the group experience as facilitative around discussions of professional ambiguity. Providing an opportunity for physicians to reflect on professional behavior in an open and safe forum may be a practical way to guide physicians to assess themselves on professional behavior and engage with the complexities of their work. The finding that the focus groups led to reflection at a group level suggests that effective reflection on professional behavior may require a socially interactive process. Emphasizing both the behaviors and the internal and external context in which they occur can thus be viewed as critically important for understanding professionalism in practicing physicians.
An Evolved International Lunar Decade Global Exploration Roadmap
NASA Astrophysics Data System (ADS)
Dunlop, D.; Holder, K.
2015-10-01
An Evolved Global Exploration Roadmap (GER) reflecting a proposed International Lunar Decade is presented by an NSS chapter to address many of the omissions and new prospective commercial mission developments since the 2013 edition of the ISECG GER.
Comparison of FRA regulations to international high-speed rail standards.
DOT National Transportation Integrated Search
2013-05-01
This report compares international standards with selected parts of Federal Railroad Administration (FRA) Code of Federal Regulation Title 49 CFR Transportation. The parts were preselected by FRA and are meant to reflect those areas of the regulation...
Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-25
... sections of the interim final guidance at 2 CFR part 176 that pertain to international agreements. These amendments reflect changes with respect to U.S. international obligations since the publication of the... international agreements. First, it makes a technical correction to section 176.90(a). Second, it changes the...
ERIC Educational Resources Information Center
Garner, Annie A.; Mrug, Sylvie; Hodgens, Bart; Patterson, Cryshelle
2013-01-01
Objective: Symptoms of sluggish cognitive tempo (SCT) are correlated with inattention and internalizing difficulties. The purpose of the present study was to determine whether symptoms of SCT reflect comorbid internalizing disorder with ADHD or a separate syndrome. Method: Using a clinical sample of youth evaluated for behavioral and learning…
ERIC Educational Resources Information Center
Schubring, Gert
2012-01-01
PME, the International Group for the Psychology of Mathematics Education, was founded in 1976, at the "Third International Congress on Mathematical Education" in Karlsruhe, organised by the International Commission on Mathematics Instruction (ICMI). While PME is thus beyond coming of age and is reflecting its further orientation--due to…
ERIC Educational Resources Information Center
McGrath, Simon; Madziva, Roda; Thondhlana, Juliet
2017-01-01
The last decade has seen the rise of literature focused on the rapid expansion of international students in higher education globally and the growing policy discourse around improving graduate employability. However, both, inevitably, have limitations. Together, they tend to homogenise international learners and see them narrowly as simply…
International Handbook of Research and Development of Giftedness and Talent.
ERIC Educational Resources Information Center
Heller, Kurt A., Ed.; And Others
This international handbook provides a synthesis and critical review of the significant theory and research dealing with all aspects of giftedness. Each article is designed to reflect the state-of-the art from an international perspective, to offer a comprehensive review, and to comprise the forefront of knowledge and thought about the gifted. The…
From Heads to Hearts: Digital Stories as Reflection Artifacts of Teachers' International Experience
ERIC Educational Resources Information Center
Walters, Lynne M.; Green, Martha R.; Wang, Liangyan; Walters, Timothy
2011-01-01
Because most American teachers work with a diverse student population, they need to be interculturally competent to interact successfully with their students. One of the best hopes for developing cultural competence is through an international experience. As important as it is for teachers to have an international experience, it is even more…
NASA Astrophysics Data System (ADS)
Fofanov, Ya A.
2009-06-01
A series of reflection resonances formed by the hyperfine components of the D2-lines in the spectrum of the natural mixture of rubidium isotopes is studied. Passages from resonantly frustrated total internal reflection to resonance Brewster reflection caused by the frequency tuning of the incident light are demonstrated experimentally. The contrast of the strongest refection resonances exceeds 500% at the moderate heating of reflecting cells. The intensity of the reflected light changes in this case by more than 20 times. A theory is developed which is based on a two-level model for resonance atoms and Fresnel formulas for reflection coefficients. Numerical calculations based on the proposed theory confirm main experimental results.
NASA Astrophysics Data System (ADS)
Grimshaw, R. H. J.; Baines, P. G.; Bell, R. C.
1985-07-01
We consider the three-dimensional reflection and diffraction properties of internal waves in a continuously stratified rotating fluid which are incident on the junction of a vertical slit and a half-space. This geometry is a model for submarine canyons on continental slopes in the ocean, where various physical phenomena embodying reflection and diffraction effects have been observed. Three types of incident wave are considered: (1) Kelvin waves in the slit (canyon); (2) Kelvin waves on the slope; and (3) plane internal waves incident from the half-space (ocean). These are scattered into Kelvin and Poincaré waves in the slit, a Kelvin wave on the slope and Poincaré waves in the half-space. Most of the discussion is centered around case (1). Various properties of the wave field are calculated for ranges of the parameters c/ cot θ, γα and ƒ/ω where cot θ is the topographic slope, c is the internal wave ray slope, α is the canyon half-width, γ is the down-slope wave-number, ƒ is the Coriolis parameter and ω is the wave frequency. Analytical results are obtained for small γα and some approximate results for larger values of γα. The results show that significant wave trapping may occur in oceanic situations, and that submarine canyons may act as source regions for internal Kelvin waves on the continental slope.
"Colgate-Plus"--An Imperfect Light Pipe.
ERIC Educational Resources Information Center
Ouseph, P. J.
1993-01-01
Describes the effects when a clear toothbrush is used as total internally reflecting light pipe when attached to a laser. Several explanations are given describing the nonuniformity of the transmitted light resulting from surface and internal irregularities found in the toothbrush. (MVL)
Verification of reflectance models in turbid waters
NASA Technical Reports Server (NTRS)
Tanis, F. J.; Lyzenga, D. R.
1981-01-01
Inherent optical parameters of very turbid waters were used to evaluate existing water reflectance models. Measured upwelling radiance spectra and Monte Carlo simulations of the radiative transfer equations were compared with results from models based upon two flow, quasi-single scattering, augmented isotropic scattering, and power series approximation. Each model was evaluated for three separate components of upwelling radiance: (1) direct sunlight; (2) diffuse skylight; and (3) internally reflected light. Limitations of existing water reflectance models as applied to turbid waters and possible applications to the extraction of water constituent information are discussed.
NASA Astrophysics Data System (ADS)
Foley, Tyler; Pegram, Matthew; Jenkins, Zachary; Hester, Brooke C.; Burris, Jennifer L.
2015-01-01
We have developed an eye-catching demonstration that showcases a variety of physics topics from total internal reflection to electrostatics to non-Newtonian fluid dynamics, including the Kaye effect. The essential components of the demonstration include a vertical stream of liquid soap in which a laser pointer is internally reflected, and which subsequently hits an inclined plane. As the liquid soap, a non-Newtonian fluid, begins to accumulate into a pile, its shear properties change and the incoming fluid rebounds from the pile, forming striking parabolic arcs. We present here a readily reproducible and inexpensive version of a laser soap fountain.
Back focal plane microscopic ellipsometer with internal reflection geometry
NASA Astrophysics Data System (ADS)
Otsuki, Soichi; Murase, Norio; Kano, Hiroshi
2013-05-01
A back focal plane (BFP) ellipsometer is presented to measure a thin film on a cover glass using an oil-immersion high-numerical-aperture objective lens. The internal reflection geometry lowers the pseudo Brewster angle (ϕB) to the range over which the light distribution is observed in BFP of the objective. A calculation based on Mueller matrix was developed to compute ellipsometric parameters from the intensity distribution on BFP. The center and radius of the partial reflection region below the critical angle were determined and used to define a polar coordinate on BFP. Harmonic components were computed from the intensities along the azimuth direction and transformed to ellipsometric parameters at multiple incident angles around ϕB. The refractive index and thickness of the film and the contributions of the objective effect were estimated at the same time by fitting.
NASA Astrophysics Data System (ADS)
Alsop, Steve; Ames, Patricia; Arroyo, Graciela Cordero; Dippo, Don
2010-12-01
This article explores distinctive features of a 5-year international education development project set in rural northern Peru (PROMEB, the Proyecto de Mejoramiento de la Educación Básica). Grounded within a partnership between teacher educators from Peru, Mexico and Canada, and rural Peruvian teachers, students and their communities, we offer reflections on a teacher education initiative which sought to support action-orientated inquiries as a mechanism for school/community development. Set against a background of poverty, hunger, isolation and an "educational crisis", we outline our pedagogy and describe two projects. We then reflect on the influences of our engagements and on associated tensions and ambiguities in our methods. We hope that such discussions might offer insights for others involved in international school/community development projects of this type.
Bringing the Real World in: Reflection on Building a Virtual Learning Environment
ERIC Educational Resources Information Center
Mundkur, Anuradha; Ellickson, Cara
2012-01-01
We reflect on translating participatory and experiential learning methodologies into an online teaching environment through a Virtual Learning Environment (VLE) that simulates the "real-world" contexts of international development in order to develop an applied critical understanding of gender analysis and gender mainstreaming. Rather than being…
[Reflections on qualitative research. Interview of Luisa Saiani with Luigina Mortari].
Mortari, Luigina; Saiani, Luisa
2013-01-01
Reflections on qualitative research. Interview of Luisa Saiani to Luigina Mortari. Luigina Mortari, an internationally known expert of epistemology and qualitative research, was interviewed to explore her thoughts on issues relevant for qualitative research: when a research question can be considered relevant; key methodological elements; ethical issues.
Becoming Cosmopolitan and Other Dilemmas of Internationalisation: Reflections from the Gulf States
ERIC Educational Resources Information Center
McNiff, Jean
2013-01-01
Drawing on the concept of cosmopolitans and locals within competing discourses regarding the aims of higher education and international marketization, this paper suggests that cultural cosmopolitanism may be developed through intercultural dialogue. It reflects on the findings of an action research-based teacher professional education programme in…
Towards Continuous Professional Development: Experiencing Group Reflection to Analyse Practice
ERIC Educational Resources Information Center
Sharmahd, Nima; Peeters, Jan; Bushati, Mirlinda
2018-01-01
Researchers and international organisations recognise that the quality of ECEC services is related to a professional and competent workforce. The latter should be part of a "competent system" that is capable of linking staff's initial good education to the possibility of constantly reflecting on ideas and practices. Continuous…
ICON: Radical Professional Development in the Conservatoire
ERIC Educational Resources Information Center
Duffy, Celia
2016-01-01
The idea for the Innovative Conservatoire (ICON) was first proposed at the Reflective Conservatoire Conference in 2006. An international collaboration which stimulates knowledge exchange, innovation and reflective practice in conservatoires, ICON has opened up an area of work that is often carried out behind closed doors. Working via creative…
Reflecting on the 5th World Environmental Education Congress, Montreal, 2009
ERIC Educational Resources Information Center
Jickling, Bob
2010-01-01
This article traces the development of the World Environmental Congress movement and its establishment as an important international forum. Reflecting on the 5th Congress, it notes the particular contribution of the Congress theme, "Our Common Home". Finally, it considers environmental education's place alongside other parallel transformative…
Polarized and non-polarized leaf reflectances of Coleus blumei
NASA Technical Reports Server (NTRS)
Grant, Lois; Daughtry, C. S. T.; Vanderbilt, V. C.
1987-01-01
A polarization photometer has been used to measure the reflectance of three variegated portions of Coleus blumei, Benth. in five wavelength bands of the visible and near-infrared spectrum. The polarized component of the reflectance factor was found to be independent of wavelength, indicating that the polarized reflectance arises from the leaf surface. It is suggested that differences in the polarized component result from variations in surface features. The nonpolarized component of the reflectance factor is shown to be related to the internal leaf structure. The variation of the degree of polarization with wavelength was found to be greatest in the regions of the spectrum where absorption occurs.
Sea monsters & whirlpools: Navigating between examination and reflection in medical education.
Hodges, Brian David
2015-03-01
The 16th International Ottawa Conference/Canadian Conference on Medical Education (2014) featured a keynote deconstructing the rising discourse of competence-as-reflection in medical education. This paper, an elaborated version of the presentation, is an investigation into the theoretical roots of the diverse forms of reflective practice that are being employed by medical educators. It also raises questions about the degree to which any of these practices is compatible with assessment.
NASA Astrophysics Data System (ADS)
Kuramoto, Masaru; Kobayashi, Seiichiro; Akagi, Takanobu; Tazawa, Komei; Tanaka, Kazufumi; Saito, Tatsuma; Takeuchi, Tetsuya
2018-03-01
We have achieved a high output power of 6 mW from a 441 nm GaN-based vertical-cavity surface-emitting laser (VCSEL) under continuous wave (CW) operation, by reducing both the internal loss and the reflectivity of the front cavity mirror. A preliminary analysis of the internal loss revealed an enormously high transverse radiation loss in a conventional GaN-based VCSEL without lateral optical confinement (LOC). Introducing an LOC structure enhanced the slope efficiency by a factor of 4.7, with a further improvement to a factor of 6.7 upon reducing the front mirror reflectivity. The result was a slope efficiency of 0.87 W/A and an external differential quantum efficiency of 32% under pulsed operation. A flip-chip-bonded VCSEL also exhibited a high slope efficiency of 0.64 W/A and an external differential quantum efficiency of 23% for the front-side output under CW operation. The reflectivity of the cavity mirror was adjusted by varying the number of AlInN/GaN distributed Bragg reflector pairs from 46 to 42, corresponding to reflectivity values from 99.8% to 99.5%. These results demonstrate that a combination of internal loss reduction and cavity mirror control is a very effective way of obtaining a high output GaN-based VCSEL.
The Rehabilitation Paradox: Street-Working Children in Afghanistan
ERIC Educational Resources Information Center
Williams, Christopher; Yazdani, Farzaneh
2009-01-01
International humanitarian intervention in Afghanistan reflects a policy discourse of "rehabilitation," which is very evident in relation to nongovernmental organization (NGO) projects for street-working children. Through analysing national and international policy, professional perceptions of the children, and field visits to see how…
The "Review" at Thirty. Editorial.
ERIC Educational Resources Information Center
Altbach, Philip G.
1986-01-01
On this journal's 30th anniversary, the editor reflects on the development of the field of comparative education and its "infrastructure" (scholars and practitioners, scholarly journals, book publishers, and international agencies); the field's internationalization; influence of international aid donors on research; symbiosis of comparative and…
ERIC Educational Resources Information Center
Lane, Kathleen Lynne; Oakes, Wendy P.; Harris, Pamela J.; Menzies, Holly Mariah; Cox, Meredith; Lambert, Warren
2012-01-01
We report findings of an exploratory validation study of a revised instrument: the Student Risk Screening Scale-Internalizing and Externalizing (SRSS-IE). The SRSS-IE was modified to include seven additional items reflecting characteristics of internalizing behaviors, with proposed items generated from the current literature base, review of…
International Policy and Roma Education in Europe: Essential Inputs or Centralized Distractions?
ERIC Educational Resources Information Center
Ram, Melanie H.
2015-01-01
This article examines the case of Roma inclusion in education as a study of the ways in which international policy can and cannot bring change at the local level. I first reflect on the numerous international organizations and initiatives to improve the educational outcomes of Roma, and to reduce segregation and other non-inclusionary practices.…
ERIC Educational Resources Information Center
Education International, Brussels (Belgium).
The papers in this volume reflect the general theme of the conference, which includes issues that will shape the future of Education International and of education for decades to come. The first half of the publication includes speeches by the following individuals: (1) Mary Hatwood Futrell, President, Education International; (2) Sandra Feldman,…
Ensink, Karin; Bégin, Michaël; Normandin, Lina; Fonagy, Peter
2017-11-01
The objective was to examine pathways from child sexual abuse (CSA) and maternal mentalizing to child internalizing and externalizing difficulties and to test a model of MRF as a moderator of the relationships between CSA and child difficulties. The sample was comprised of 154 mothers and children aged 2-12 where 64 children had experienced CSA. To assess parental mentalizing the Parental Development Interview was rated with the Parental Reflective Functioning Scale. Child internalizing and externalizing difficulties were assessed with the Child Behavior Checklist (CBCL). Results indicate that there were significant inverse relationships between maternal mentalizing and child internalizing and externalizing difficulties. When maternal mentalizing was considered together with CSA, only maternal mentalizing was a significant predictor of child difficulties. Furthermore, maternal mentalizing moderated the relationship between CSA and child internalizing difficulties. These findings provide evidence of the importance of the parents' mentalizing stance for psychiatric symptoms of children aged 2-12, as well as children's recovery from CSA. The clinical implications of the findings are discussed. Copyright © 2017 Elsevier B.V. All rights reserved.
A framework to teach self-reflection for the remedial resident.
Leung, Fok-Han; Ratnapalan, Savithri
2011-01-01
Regardless of the area of deficiency, be it in knowledge, skills or attitudes, residents requiring remediation are rarely self-identified. This illustrates a diminished ability for self-reflection. Self-reflection is a cornerstone of adult education. During the remediation process, the remediation curriculum needs to emphasize self-reflection. How can one structure self-reflection in a remediation curriculum? This article describes how to adapt and apply environmental scanning for remedial residents. Environmental scanning is a rigorous and well-developed business approach that can be adapted for personal continuous quality improvement to foster self-reflection in medical trainees. There are often already existing tools which can form the foundation for regular reflection in medical education using an environmental scanning structure. Environmental scanning can be thought of as a structured approach to internal and external reflections.
Planar Reflection of Gaseous Detonations
NASA Astrophysics Data System (ADS)
Damazo, Jason Scott
Pipes containing flammable gaseous mixtures may be subjected to internal detonation. When the detonation normally impinges on a closed end, a reflected shock wave is created to bring the flow back to rest. This study built on the work of Karnesky (2010) and examined deformation of thin-walled stainless steel tubes subjected to internal reflected gaseous detonations. A ripple pattern was observed in the tube wall for certain fill pressures, and a criterion was developed that predicted when the ripple pattern would form. A two-dimensional finite element analysis was performed using Johnson-Cook material properties; the pressure loading created by reflected gaseous detonations was accounted for with a previously developed pressure model. The residual plastic strain between experiments and computations was in good agreement. During the examination of detonation-driven deformation, discrepancies were discovered in our understanding of reflected gaseous detonation behavior. Previous models did not accurately describe the nature of the reflected shock wave, which motivated further experiments in a detonation tube with optical access. Pressure sensors and schlieren images were used to examine reflected shock behavior, and it was determined that the discrepancies were related to the reaction zone thickness extant behind the detonation front. During these experiments reflected shock bifurcation did not appear to occur, but the unfocused visualization system made certainty impossible. This prompted construction of a focused schlieren system that investigated possible shock wave-boundary layer interaction, and heat-flux gauges analyzed the boundary layer behind the detonation front. Using these data with an analytical boundary layer solution, it was determined that the strong thermal boundary layer present behind the detonation front inhibits the development of reflected shock wave bifurcation.
The United States and International Music Education.
ERIC Educational Resources Information Center
Werner, Robert J.
1990-01-01
Outlines music education's history since World War II, highlighting the work of the Music Educators National Conference (MENC). Emphasizes U.S. music educators' influence in international music education, arguing this influence should reflect multicultural, global perspectives. Notes the importance of understanding mass media's influence on young…
International Immersion in Counselor Education: A Consensual Qualitative Research Investigation
ERIC Educational Resources Information Center
Barden, Sejal M.; Cashwell, Craig S.
2014-01-01
This study used consensual qualitative research methodology to examine the phenomenon of international immersion on counselor education students' (N = 10) development and growth. Seven domains emerged from the data (cultural knowledge, empathy, personal and professional impact, process/reflection, relationships, personal characteristics, and…
ERIC Educational Resources Information Center
Cullen, Rowena; Nagata, Haruki
2008-01-01
Academic libraries in Japan are well resourced by international standards, and support Japan's internationally recognized research capability well, but there are also ways in which they reflect Japan's strong bureaucratic culture. Recent changes to the status of national university libraries have seen a new interest in customer service, and…
Interactive calculation procedures for mixed compression inlets
NASA Technical Reports Server (NTRS)
Reshotko, Eli
1983-01-01
The proper design of engine nacelle installations for supersonic aircraft depends on a sophisticated understanding of the interactions between the boundary layers and the bounding external flows. The successful operation of mixed external-internal compression inlets depends significantly on the ability to closely control the operation of the internal compression portion of the inlet. This portion of the inlet is one where compression is achieved by multiple reflection of oblique shock waves and weak compression waves in a converging internal flow passage. However weak these shocks and waves may seem gas-dynamically, they are of sufficient strength to separate a laminar boundary layer and generally even strong enough for separation or incipient separation of the turbulent boundary layers. An understanding was developed of the viscous-inviscid interactions and of the shock wave boundary layer interactions and reflections.
25 CFR 542.19 - What are the minimum internal control standards for accounting?
Code of Federal Regulations, 2013 CFR
2013-04-01
...; (3) Individual and statistical game records to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop by each table game, and to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop for each type of table game, by...
25 CFR 542.19 - What are the minimum internal control standards for accounting?
Code of Federal Regulations, 2014 CFR
2014-04-01
...; (3) Individual and statistical game records to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop by each table game, and to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop for each type of table game, by...
25 CFR 542.19 - What are the minimum internal control standards for accounting?
Code of Federal Regulations, 2010 CFR
2010-04-01
...; (3) Individual and statistical game records to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop by each table game, and to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop for each type of table game, by...
25 CFR 542.19 - What are the minimum internal control standards for accounting?
Code of Federal Regulations, 2011 CFR
2011-04-01
...; (3) Individual and statistical game records to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop by each table game, and to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop for each type of table game, by...
25 CFR 542.19 - What are the minimum internal control standards for accounting?
Code of Federal Regulations, 2012 CFR
2012-04-01
...; (3) Individual and statistical game records to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop by each table game, and to reflect statistical drop, statistical win, and the percentage of statistical win to statistical drop for each type of table game, by...
Improving Intercultural Competence in the Classroom: A Reflective Development Model
ERIC Educational Resources Information Center
Feng, Jing Betty
2016-01-01
To meet the increased demand for international business education that prepares college students for studying, living, or working overseas, I propose a four-stage reflective development model to be used in the traditional classroom context to enhance intercultural competence for undergraduate students. I employ the model in a personal development…
Internal Reflection Spectra of Surface Compounds and Adsorbed Molecules
NASA Astrophysics Data System (ADS)
Zolotarev, V. M.; Lygin, V. I.; Tarasevich, B. N.
1981-01-01
The application of attenuated total reflection (ATR) spectroscopy in surface studies of inorganic adsorbents and catalysts, polymers, and optically transparent electrodes is discussed. The basic principles of ATR spectroscopy as applied to surface phenomena are considered, with special reference to thin films, industrial adsorbents and catalysts, and polymer degradation processes. 276 references.
Teachers' Reflections on Education in a Global Age
ERIC Educational Resources Information Center
Callis, Laura Kyser; Osborn, Daniel
2014-01-01
This article presents profiles of and reflections by teachers with international experience, including the authors, who offer insights on education in a global age. The respondents who were colleagues of the authors were interviewed to learn about their K-12 education, insights into and analysis of their experiences teaching abroad, and thoughts…
The Quest for World Class Universities in China: Critical Reflections
ERIC Educational Resources Information Center
Ngok, Kinglun; Guo, Weiqing
2008-01-01
Building world-class universities has become a national policy priority in China since then-President Jiang Zemin announced in May 1998 that China must have several world-class universities of international advanced level. This article aims to offer critical reflections on the policy in relation to building world-class universities in China. It…
The Implementation of Progress Files in Higher Education: Reflection as National Policy
ERIC Educational Resources Information Center
Clegg, Sue; Bradley, Sally
2006-01-01
Progress files, involving personal development planning (PDP), are becoming a feature of many higher education systems internationally. In the UK they will become mandatory for all undergraduate students from 2005. This presents a major implementation challenge, because while reflection has been a cornerstone of practice in some areas of higher…
Pitch-catch only ultrasonic fluid densitometer
Greenwood, M.S.; Harris, R.V.
1999-03-23
The present invention is an ultrasonic fluid densitometer that uses a material wedge and pitch-catch only ultrasonic transducers for transmitting and receiving ultrasonic signals internally reflected within the material wedge. Density of a fluid is determined by immersing the wedge into the fluid and measuring reflection of ultrasound at the wedge-fluid interface. 6 figs.
Pitch-catch only ultrasonic fluid densitometer
Greenwood, Margaret S.; Harris, Robert V.
1999-01-01
The present invention is an ultrasonic fluid densitometer that uses a material wedge and pitch-catch only ultrasonic transducers for transmitting and receiving ultrasonic signals internally reflected within the material wedge. Density of a fluid is determined by immersing the wedge into the fluid and measuring reflection of ultrasound at the wedge-fluid interface.
Fifty Years of "Comparative Education Review" Bibliographies: Reflections on the Field
ERIC Educational Resources Information Center
Raby, Rosalind Latiner
2007-01-01
For the past three years, the "Comparative Education Review" ("CER") has published annual bibliographic essays reflecting the changes in the field over the previous year. This year, to mark the 50th anniversary of the Comparative and International Education Society (CIES), the author analyzed "CER" bibliographies since 1959, in addition to…
Iterated intracochlear reflection shapes the envelopes of basilar-membrane click responses
Shera, Christopher A.
2015-01-01
Multiple internal reflection of cochlear traveling waves has been argued to provide a plausible explanation for the waxing and waning and other temporal structures often exhibited by the envelopes of basilar-membrane (BM) and auditory-nerve responses to acoustic clicks. However, a recent theoretical analysis of a BM click response measured in chinchilla concludes that the waveform cannot have arisen via any equal, repetitive process, such as iterated intracochlear reflection [Wit and Bell (2015), J. Acoust. Soc. Am. 138, 94–96]. Reanalysis of the waveform contradicts this conclusion. The measured BM click response is used to derive the frequency-domain transfer function characterizing every iteration of the loop. The selfsame transfer function that yields waxing and waning of the BM click response also captures the spectral features of ear-canal stimulus-frequency otoacoustic emissions measured in the same animal, consistent with the predictions of multiple internal reflection. Small shifts in transfer-function phase simulate results at different measurement locations and reproduce the heterogeneity of BM click response envelopes observed experimentally. PMID:26723327
Attention to memory: orienting attention to sound object representations.
Backer, Kristina C; Alain, Claude
2014-01-01
Despite a growing acceptance that attention and memory interact, and that attention can be focused on an active internal mental representation (i.e., reflective attention), there has been a paucity of work focusing on reflective attention to 'sound objects' (i.e., mental representations of actual sound sources in the environment). Further research on the dynamic interactions between auditory attention and memory, as well as its degree of neuroplasticity, is important for understanding how sound objects are represented, maintained, and accessed in the brain. This knowledge can then guide the development of training programs to help individuals with attention and memory problems. This review article focuses on attention to memory with an emphasis on behavioral and neuroimaging studies that have begun to explore the mechanisms that mediate reflective attentional orienting in vision and more recently, in audition. Reflective attention refers to situations in which attention is oriented toward internal representations rather than focused on external stimuli. We propose four general principles underlying attention to short-term memory. Furthermore, we suggest that mechanisms involved in orienting attention to visual object representations may also apply for orienting attention to sound object representations.
Strategy to Enhance International Supply Chain Security
2007-07-01
as part of the effort to secure air passenger travel . The security assessment crew traveling by air, land or sea cannot be considered only a travel ...threats through its traveler screening and worker credentialing programs. The strategy to secure the supply chain reflects the larger security strategy of...living or traveling abroad. • Assisting U.S. businesses in the international marketplace. • Coordinating and providing support for international
Astronomers Travel in Time and Space with Light
NASA Technical Reports Server (NTRS)
Mather, John C.
2016-01-01
This is an excerpt of John Mather's in a book titled: INSPIRED BY LIGHT, Reflections from the International Year of Light 2015. It was produced in January 2016 by SPIE, the European Physical Society (EPS), and The Abdus Salam International Centre for Theoretical Physics (ICTP) to commemorate the International Year of Light and Light-based Technologies 2015. The excerpt discusses how astronomers use light.
NASA Technical Reports Server (NTRS)
Siegel, Robert
1996-01-01
Surface convection and refractive index are examined during transient radiative heating or cooling of a grey semitransparent layer with internal absorption, emission and conduction. Each side of the layer is exposed to hot or cold radiative surroundings, while each boundary is heated or cooled by convection. Emission within the layer and internal reflections depend on the layer refractive index. The reflected energy and heat conduction distribute energy across the layer and partially equalize the transient temperature distributions. Solutions are given to demonstrate the effect of radiative heating for layers with various optical thicknesses, the behavior of the layer heated by radiation on one side and convectively cooled on the other, and a layer heated by convection while being cooled by radiation. The numerical method is an implicit finite difference procedure with non-uniform space and time increments. The basic method developed in earlier work is expanded to include external convection and incident radiation.
Xie, Peng; Lin, Huichuan; Liu, Yong; Li, Baojun
2014-10-20
We present a waveguide coupling approach for planar waveguide solar concentrator. In this approach, total internal reflection (TIR)-based symmetric air prisms are used as couplers to increase the coupler reflectivity and to maximize the optical efficiency. The proposed concentrator consists of a line focusing cylindrical lens array over a planar waveguide. The TIR-based couplers are located at the focal line of each lens to couple the focused sunlight into the waveguide. The optical system was modeled and simulated with a commercial ray tracing software (Zemax). Results show that the system used with optimized TIR-based couplers can achieve 70% optical efficiency at 50 × geometrical concentration ratio, resulting in a flux concentration ratio of 35 without additional secondary concentrator. An acceptance angle of ± 7.5° is achieved in the x-z plane due to the use of cylindrical lens array as the primary concentrator.
Australia and international treaties: population.
Hugo, G
1995-05-01
"The separation of environmental and population issues globally is reflected in the fact that the setting of international agendas to tackle them was for all intents and purposes undertaken at separate international meetings--the Earth Summit in Rio de Janeiro in 1992 and the International Population Conference in Cairo in 1994. This paper presents an overview of Australia's international involvement in population issues and suggests that until recently Australia had played a minor political role in international population initiatives primarily, and perhaps ironically, because it has been a world leader in demographic research and teaching for several decades." excerpt
An update on X-ray reflection gratings developed for future missions
NASA Astrophysics Data System (ADS)
Miles, Drew
2018-01-01
X-ray reflection gratings are a key technology being studied for future X-ray spectroscopy missions, including the Lynx X-ray mission under consideration for the 2020 Decadal Survey. We present an update on the status of X-ray reflection gratings being developed at Penn State University, including current fabrication techniques and mass-replication processes and the latest diffraction efficiency results and resolving power measurements. Individual off-plane X-ray reflection gratings have exceeded the current Lynx requirements for both effective area and resolving power. Finally, we discuss internal projects that will advance the technology readiness level of these gratings.
Kim, Iltai; Kihm, Kenneth D
2010-02-01
Time-dependent and near-field nanoparticle concentrations are determined by correlating the surface plasmon resonance (SPR) reflectance intensities with the effective refractive index (ERI) of the nanofluid under evaporation. A critical angle measurement for total internal reflection identifies the ERI of the nanofluid at different nanoparticle concentrations. The corresponding SPR reflectance intensities correlate the nanofluidic ERI with the nanoparticle concentrations. Example applications for evaporating nanofluidic droplets containing 47 nmAl(2)O(3) particles demonstrate the feasibility of this new imaging tool for measuring time-resolved and full-field nanoparticle concentration profiles.
48 CFR 52.216-7 - Allowable Cost and Payment.
Code of Federal Regulations, 2011 CFR
2011-10-01
... last disclosure of internal audit reports to the Government. (J) Annual internal audit plan of...-contract cost allowability limitations, and billing limitations. (v) The Contractor shall update the billings on all contracts to reflect the final settled rates and update the schedule of cumulative direct...
International Service-Learning: Students' Personal Challenges and Intercultural Competence
ERIC Educational Resources Information Center
Nickols, Sharon Y.; Rothenberg, Nancy J.; Moshi, Lioba; Tetloff, Meredith
2013-01-01
International service-learning offers students a complex cluster of educational opportunities that include cultural competency acquisition combined with professional development. An interdisciplinary program in a remote area of Tanzania revealed that the journey toward competence can be an arduous one. Drawing from students' reflections in…
Grammaticality and Educational Research
ERIC Educational Resources Information Center
Hordern, Jim
2017-01-01
This paper uses Bernstein's concept of grammar to illuminate aspects of educational research. The relationship between internal and external languages of description in the production of disciplinary knowledge is examined. This leads to a reflection on the various factors both internal and external to the discipline of educational studies that…
26 CFR 1.446-6 - REMIC inducement fees.
Code of Federal Regulations, 2011 CFR
2011-04-01
... Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Methods of Accounting § 1.446-6 REMIC inducement fees. (a) Purpose. This section provides specific timing rules for the clear reflection of income from an inducement fee received in...
Contracting and Purchasing Management in the International Marketplace
1989-09-01
Fortune magazine entitled "B- Schools Get a Global Vision" explained how many of the top graduate business schools are restructuring their curriculums...become critical and more complex in the international environment. Many top graduate business schools are changing their curricula to reflect a more
ERIC Educational Resources Information Center
Ballantyne, Roy; Uzzell, David
1999-01-01
In the past decade, heritage and environmental interpretation shifted from technical emphasis to social perspective, reflected in five international trends: growing concern with theory, increasing ecotourism and consequent changes in environmental interpretation, reactions to globalization (homogeneity versus uniqueness), interpretation of…
ERIC Educational Resources Information Center
United Nations Educational, Scientific, and Cultural Organization, Hamburg (Germany). Inst. for Education.
These proceedings contain the following papers: "The International Seminar on Women's Empowerment" (Anonuevo, Bochynek); "The Theoretical and Practical Bases for Empowerment" (Stromquist); "Some Reflections on the Empowerment of Women" (Lazo); "Women's Literacy and Empowerment: The Nellore Experience"…
Mass-Customisation and Self-Reflective Frameworks: Early Developments in New Zealand
ERIC Educational Resources Information Center
Clayton, John
2012-01-01
Education has long been regarded as the foundation stone of national growth and international competitiveness. In the last three decades national educational reforms to improve access to higher education qualifications, individual higher education institutions' aggressive national and international marketing initiatives and improved information…
Air pollution and public health: a guidance document for risk managers.
Craig, Lorraine; Brook, Jeffrey R; Chiotti, Quentin; Croes, Bart; Gower, Stephanie; Hedley, Anthony; Krewski, Daniel; Krupnick, Alan; Krzyzanowski, Michal; Moran, Michael D; Pennell, William; Samet, Jonathan M; Schneider, Jurgen; Shortreed, John; Williams, Martin
2008-01-01
This guidance document is a reference for air quality policymakers and managers providing state-of-the-art, evidence-based information on key determinants of air quality management decisions. The document reflects the findings of five annual meetings of the NERAM (Network for Environmental Risk Assessment and Management) International Colloquium Series on Air Quality Management (2001-2006), as well as the results of supporting international research. The topics covered in the guidance document reflect critical science and policy aspects of air quality risk management including i) health effects, ii) air quality emissions, measurement and modeling, iii) air quality management interventions, and iv) clean air policy challenges and opportunities.
Nonpolarizing beam splitter designed by frustrated total internal reflection inside a glass cube.
Xu, Xueke; Shao, Jianda; Fan, Zhengxiu
2006-06-20
A method for the design of an all-dielectric nonpolarizing prism beam splitter utilizing the principle of frustrated total internal reflection is reported. The nonpolarizing condition for a prism beam splitter is discussed, and some single layer design examples are elaborated. The concept can be applied to a wide range of wavelengths and arbitrary transmittance values, and with the help of a computer design program examples of 400-700 nm, T(p)=T(s)=0.5+/-0.01, with incident angles of 45 degrees and 62 degrees are given. In addition, the sensitivity and application of the design are also discussed.
NASA Technical Reports Server (NTRS)
Blumberg, Seth; Gajraj, Arivalagan; Pennington, Matthew W.; Meiners, Jens-Christian
2005-01-01
Tethered particle microscopy is a powerful tool to study the dynamics of DNA molecules and DNA-protein complexes in single-molecule experiments. We demonstrate that stroboscopic total internal reflection microscopy can be used to characterize the three-dimensional spatiotemporal motion of DNA-tethered particles. By calculating characteristic measures such as symmetry and time constants of the motion, well-formed tethers can be distinguished from defective ones for which the motion is dominated by aberrant surface effects. This improves the reliability of measurements on tether dynamics. For instance, in observations of protein-mediated DNA looping, loop formation is distinguished from adsorption and other nonspecific events.
Broad Band Intra-Cavity Total Reflection Chemical Sensor
Pipino, Andrew C. R.
1998-11-10
A broadband, ultrahigh-sensitivity chemical sensor is provided that allows etection through utilization of a small, extremely low-loss, monolithic optical cavity. The cavity is fabricated from highly transparent optical material in the shape of a regular polygon with one or more convex facets to form a stable resonator for ray trajectories sustained by total internal reflection. Optical radiation enters and exits the monolithic cavity by photon tunneling in which two totally reflecting surfaces are brought into close proximity. In the presence of absorbing material, the loss per pass is increased since the evanescent waves that exist exterior to the cavity at points where the circulating pulse is totally reflected, are absorbed. The decay rate of an injected pulse is determined by coupling out an infinitesimal fraction of the pulse to produce an intensity-versus-time decay curve. Since the change in the decay rate resulting from absorption is inversely proportional to the magnitude of absorption, a quantitative sensor of concentration or absorption cross-section with 1 part-per-million/pass or better sensitivity is obtained. The broadband nature of total internal reflection permits a single device to be used over a broad wavelength range. The absorption spectrum of the surrounding medium can thereby be obtained as a measurement of inverse decay time as a function of wavelength.
Bohannon, Kevin P; Holz, Ronald W; Axelrod, Daniel
2017-10-01
The refractive index in the interior of single cells affects the evanescent field depth in quantitative studies using total internal reflection (TIR) fluorescence, but often that index is not well known. We here present method to measure and spatially map the absolute index of refraction in a microscopic sample, by imaging a collimated light beam reflected from the substrate/buffer/cell interference at variable angles of incidence. Above the TIR critical angle (which is a strong function of refractive index), the reflection is 100%, but in the immediate sub-critical angle zone, the reflection intensity is a very strong ascending function of incidence angle. By analyzing the angular position of that edge at each location in the field of view, the local refractive index can be estimated. In addition, by analyzing the steepness of the edge, the distance-to-substrate can be determined. We apply the technique to liquid calibration samples, silica beads, cultured Chinese hamster ovary cells, and primary culture chromaffin cells. The optical technique suffers from decremented lateral resolution, scattering, and interference artifacts. However, it still provides reasonable results for both refractive index (~1.38) and for distance-to-substrate (~150 nm) for the cells, as well as a lateral resolution to about 1 µm.
Harmonizing Landsat and Sentinel-2 Reflectances for Better Land Monitoring
NASA Technical Reports Server (NTRS)
Masek, Jeffrey; Vermote, Eric; Franch, Belen; Roger, Jean-Claude; Skakun, Sergii; Claverie, Martin; Dungan, Jennifer
2016-01-01
When combined, Landsat and ESA Sentinel-2 observations can provide 2-4 day coverage for the global land area. A collaboration among NASA GSFC (Goddard Space Flight Center), University of Maryland, and NASA Ames has developed a processing chain to create seamless, "harmonized" reflectance products using standardized atmospheric correction, BRDF (Bidirectional Reflectance Distribution Function) adjustment, spectral bandpass adjustment, and gridding algorithms. These products point the way to a "30-m MODIS (Moderate Resolution Imaging Spectroradiometer)" capability for agricultural and ecosystem monitoring by leveraging international sensors.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005022 (20 Nov. 2013) --- At a window in the International Space Station?s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth?s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station?s power supply. The light reflection phenomenon is measured in units called albedo.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005014 (20 Nov. 2013) --- At a window in the International Space Station’s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth’s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station’s power supply. The light reflection phenomenon is measured in units called albedo.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005023 (20 Nov. 2013) --- At a window in the International Space Station?s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth?s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station?s power supply. The light reflection phenomenon is measured in units called albedo.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005031 (20 Nov. 2013) --- At a window in the International Space Station?s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth?s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station?s power supply. The light reflection phenomenon is measured in units called albedo.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005016 (20 Nov. 2013) --- At a window in the International Space Station?s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth?s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station?s power supply. The light reflection phenomenon is measured in units called albedo.
Kotov during Albedo Experiment in the SM
2013-11-18
ISS038-E-005019 (20 Nov. 2013) --- At a window in the International Space Station?s Zvezda Service Module, Russian cosmonaut Oleg Kotov, Expedition 38 commander, uses a digital camera photospectral system to perform a session for the Albedo Experiment. The experiment measures Earth?s albedo, or the amount of solar radiation reflected from the surface, in the hopes to develop methods to harness the reflected radiation to supplement the station?s power supply. The light reflection phenomenon is measured in units called albedo.
Pedagogical Reflections on Internalizing Geopolitical Representations in Print Media
ERIC Educational Resources Information Center
Abdul-Jabbar, Wisam Khalid
2015-01-01
This essay explores how print media conceals implicit hegemonic texts that common readers unsuspectingly tend to internalize. These geopolitically infused texts are set to appropriate the reader's worldviews by sublimating the kind of perceptions and notions they want to promote. This paper raises questions and awareness about how academia…
International Practica Experiences as Events of Influence in a Teacher Candidates' Development
ERIC Educational Resources Information Center
Maynes, Nancy; Allison, John; Julien-Schultz, Lynn
2012-01-01
Experience may influence beliefs and beliefs may influence practices. Following these premises, we investigated teacher candidates' post experience reflections nine months after an international practicum where they taught for three weeks in rural Kenya. Teacher candidates were placed in non-governmental organization (NGO) sponsored schools on the…
International Field Experience--What Do Student Teachers Learn?
ERIC Educational Resources Information Center
Lee, Jackie Fung King
2011-01-01
This inquiry aimed to examine the benefits of having international field experience for a group of Hong Kong postgraduate student teachers who joined a six-week immersion programme in New Zealand. Through participants' reflections, interviews and programme evaluations, the present investigation found that the overseas field experience not only…
Two-pass-internal second-harmonic generation using a prism coupler.
NASA Technical Reports Server (NTRS)
Gonzalez, D. G.; Nieh, S. T. K.; Steier, W. H.
1973-01-01
A dispersive quartz prism is used to couple the total second harmonic generated in both directions by an internal cavity frequency doubler. The study shows that the dispersion of air and mirror reflection phase shifts can be compensated for by a slight nonphase match condition in the doubler.
26 CFR 1.471-1 - Need for inventories.
Code of Federal Regulations, 2010 CFR
2010-04-01
... Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Inventories § 1.471-1 Need for inventories. In order to reflect taxable income correctly... or partly finished goods and, in the case of raw materials and supplies, only those which have been...
26 CFR 1.543-1 - Personal holding company income.
Code of Federal Regulations, 2011 CFR
2011-04-01
... prevent evasion of taxes or clearly to reflect the income of any such organizations, trades, or businesses... 1.543-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Personal Holding Companies § 1.543-1 Personal holding company income. (a...
Spatial Theorizing in Comparative and International Education Research
ERIC Educational Resources Information Center
Larsen, Marianne A.; Beech, Jason
2014-01-01
The authors argue for a critical spatial perspective in comparative and international education. We briefly summarize how time and space have been conceptualized within our field. We then review mainstream social science literature that reflects a metanarrative, which we critique for contributing to false dichotomies between space and place and…
76 FR 30001 - Amendment to the International Traffic in Arms Regulations: Libya
Federal Register 2010, 2011, 2012, 2013, 2014
2011-05-24
... International Traffic in Arms Regulations: Libya AGENCY: Department of State. ACTION: Final rule. SUMMARY: The... regarding Libya to reflect the United Nations Security Council arms embargoes adopted in February and March...: (202) 261-8199; or e-mail: [email protected] . Attn: Part 126, Libya. SUPPLEMENTARY INFORMATION: On...
Journaling and Self-Study in an International Research Collective
ERIC Educational Resources Information Center
Makaiau, Amber Strong; Leng, Lu; Fukui, Suguru
2015-01-01
Drawing from the theoretical foundations of reflective teaching, culturally responsive education, social constructivism, and self-study, this collaborative self-study investigates the role of an interactive online journal in an international research collective. Each from a different country, the authors came together through a common interest in…
The Edumometer: The Commodification of Learning from Galton to the PISA
ERIC Educational Resources Information Center
Corbett, Michael
2008-01-01
This essay is a critical reflection on educational standardization, particularly the standardization of assessment flowing down from major international assessment projects like the OECD's Project for International Student Assessment to national and sub-national school systems. I argue that the establishment of an increasingly uniform metric to…
Promises from Afar: A Model of International Student Psychological Contract in Business Education
ERIC Educational Resources Information Center
Bordia, Sarbari; Bordia, Prashant; Restubog, Simon Lloyd D.
2015-01-01
Despite their significant presence in western business schools, the needs and experiences of international students have not been adequately reflected in the business education literature. We draw upon psychological contract theory--used to understand employer-employee relationships--to develop a novel theoretical model on the international…
Cautions about Inferences from International Assessments: The Case of PISA 2009
ERIC Educational Resources Information Center
Ercikan, Kadriye; Roth, Wolff-Michael; Asil, Mustafa
2015-01-01
Background/Context: Two key uses of international assessments of achievement have been (a) comparing country performances for identifying the countries with the best education systems and (2) generating insights about effective policy and practice strategies that are associated with higher learning outcomes. Do country rankings really reflect the…
Accompanying Partners of International Students: Reflections on Three Issues
ERIC Educational Resources Information Center
Cui, Dan; Arthur, Nancy; Domene, José F.
2017-01-01
This paper reviews and critiques the existing literature on accompanying partners of international students (APIS), who are often an ignored population in programs and services for the internationalization of Canadian higher education. Particularly, we identify three issues. First, we argue that current research on this group overwhelmingly…
Factors Relating to Student Grade Obsession: A Quantitative Correlational Study
ERIC Educational Resources Information Center
Thomas, Jacqueline
2011-01-01
The intense focus of students on grades attained in academia reflects a contemporary issue in higher education. The internal pressure that students experience regarding academics fuels student grade obsession. Researchers suggest that in addition to internal pressure, financial anxiety, the need to receive academic recognition and parental…
Examining Human Rights in a Global Context.
ERIC Educational Resources Information Center
Francis, Greg; Inoue, Keiko; Orrick, Stefanie
The United Nations' founding in 1945 and the 1948 adoption of the Universal Declaration of Human Rights reflected the international community's growing commitment to the protection and recognition of what is now referred to as human rights. Despite increased international attention, human rights violations continue to occur at the local, regional,…
Global Expansion among U.S. Universities: The Imperative to Examine Our Motives
ERIC Educational Resources Information Center
Birbeck, Gretchen L.
2014-01-01
The growing enthusiasm for international engagement among U.S. universities reflects the great potential gains that participation offers to both U.S. and international partners. To ensure that the benefits of such partnerships are achieved, potential participants must carefully examine and explicitly discuss their personal and institutional…
Design of an fMRI-compatible optical touch stripe based on frustrated total internal reflection.
Jarrahi, Behnaz; Wanek, Johann
2014-01-01
Previously we developed a low-cost, multi-configurable handheld response system, using a reflective-type intensity modulated fiber-optic sensor (FOS) to accurately gather participants' behavioral responses during functional magnetic resonance imaging (fMRI). Inspired by the popularity and omnipresence of the fingertip-based touch sensing user interface devices, in this paper we present the design of a prototype fMRI-compatible optical touch stripe (OTS) as an alternative configuration. The prototype device takes advantage of a proven frustrated total internal reflection (FTIR) technique. By using a custom-built wedge-shaped optically transparent acrylic prism as an optical waveguide, and a plano-concave lens to provide the required light beam profile, the position of a fingertip touching the surface of the wedge prism can be determined from the deflected light beams that become trapped within the prism by total internal reflection. To achieve maximum sensitivity, the optical design of the wedge prism and lens were optimized through a series of light beam simulations using WinLens 3D Basic software suite. Furthermore, OTS performance and MRI-compatibility were assessed on a 3.0 Tesla MRI scanner running echo planar imaging (EPI) sequences. The results show that the OTS can detect a touch signal at high spatial resolution (about 0.5 cm), and is well suited for use within the MRI environment with average time-variant signal-to-noise ratio (tSNR) loss < 3%.
Lagrangian flows within reflecting internal waves at a horizontal free-slip surface
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zhou, Qi, E-mail: q.zhou@damtp.cam.ac.uk; Diamessis, Peter J.
In this paper sequel to Zhou and Diamessis [“Reflection of an internal gravity wave beam off a horizontal free-slip surface,” Phys. Fluids 25, 036601 (2013)], we consider Lagrangian flows within nonlinear internal waves (IWs) reflecting off a horizontal free-slip rigid lid, the latter being a model of the ocean surface. The problem is approached both analytically using small-amplitude approximations and numerically by tracking Lagrangian fluid particles in direct numerical simulation (DNS) datasets of the Eulerian flow. Inviscid small-amplitude analyses for both plane IWs and IW beams (IWBs) show that Eulerian mean flow due to wave-wave interaction and wave-induced Stokes driftmore » cancels each other out completely at the second order in wave steepness A, i.e., O(A{sup 2}), implying zero Lagrangian mean flow up to that order. However, high-accuracy particle tracking in finite-Reynolds-number fully nonlinear DNS datasets from the work of Zhou and Diamessis suggests that the Euler-Stokes cancelation on O(A{sup 2}) is not complete. This partial cancelation significantly weakens the mean Lagrangian flows but does not entirely eliminate them. As a result, reflecting nonlinear IWBs produce mean Lagrangian drifts on O(A{sup 2}) and thus particle dispersion on O(A{sup 4}). The above findings can be relevant to predicting IW-driven mass transport in the oceanic surface and subsurface region which bears important observational and environmental implications, under circumstances where the effect of Earth rotation can be ignored.« less
ERIC Educational Resources Information Center
Smith, David G.
Delegates to the Canadian planning meeting of the International Year of the Child showed most interest in attending workshops on the topic of children in the international context (rather than in a single country context) and least interest in the topic of research. These choices may reflect a realization that (1) if we are to understand our own…
ERIC Educational Resources Information Center
Kim, Dongbin; Roh, Jin-young
2017-01-01
This study explores whether patterns of doctorate attainment among Chinese and Korean international students in the USA have changed over time, both in terms of quantity (i.e., number of international students) and quality (i.e., the selectivity of the undergraduate and doctoral institutions), and whether these changes reflect the improvements in…
A Summary of the Naval Postgraduate School Research Programs and Recent Publications.
1991-09-01
publication by the ASNE accepted on an international basis as an effective Naval Engineering Journal after final editing, means of reducing ship roll...Teleoperator," SUMMARY: A non-anthropomorphic, force International Journal of Robotics and Autlonomouts reflecting telemanipulator was procured in FY91...and From 22-24 May 1991. Centrifugal Instabilities," International Journal of Heat and Fluid Flow, forthcoming. THESES DIRECTED: Hughes, R.E., LT, USN
ERIC Educational Resources Information Center
Matthews, Blair
2017-01-01
International students at universities away from their home context experience a significant change to the way they engage with the world, as they think, reflect and act in response to the new context. Drawing on Archer's concept of reflexivity (2003; 2007; 2012), this paper demonstrates that international students are compelled into reflexive…
The Role of TLR2, TLR4, and TLR9 in the Pathogenesis of Atherosclerosis
2016-01-01
Toll-like receptors (TLRs) are key players in the pathogenesis of inflammatory conditions including coronary arterial disease (CAD). They are expressed by a variety of immune cells where they recognize pathogen-associated molecular patterns (PAMPs). TLRs recruit adaptor molecules, including myeloid differentiation primary response protein (MYD88) and TIRF-related adaptor protein (TRAM), to mediate activation of MAPKs and NF-kappa B pathways. They are associated with the development of CAD through various mechanisms. TLR4 is expressed in lipid-rich and atherosclerotic plaques. In TLR2−/− and TLR4−/− mice, atherosclerosis-associated inflammation was diminished. Moreover, TLR2 and TLR4 may induce expression of Wnt5a in advanced staged atheromatous plaque leading to activation of the inflammatory processes. TLR9 is activated by CpG motifs in nucleic acids and have been implicated in macrophage activation and the uptake of oxLDL from the circulation. Furthermore, TLR9 also stimulates interferon-α (INF-α) secretion and increases cytotoxic activity of CD4+ T-cells towards coronary artery tunica media smooth muscle cells. This review outlines the pathophysiological role of TLR2, TLR4, and TLR9 in atherosclerosis, focusing on evidence from animal models of the disease. PMID:27795867
Agonist-activated Ca2+ influx occurs at stable plasma membrane and endoplasmic reticulum junctions
Treves, Susan; Vukcevic, Mirko; Griesser, Johanna; Armstrong, Clara-Franzini; Zhu, Michael X.; Zorzato, Fancesco
2010-01-01
Junctate is a 33 kDa integral protein of sarco(endo)plasmic reticulum membranes that forms a macromolecular complex with inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] receptors and TRPC3 channels. TIRF microscopy shows that junctate enhances the number of fluorescent puncta on the plasma membrane. The size and distribution of these puncta are not affected by the addition of agonists that mobilize Ca2+ from Ins(1,4,5)P3-sensitive stores. Puncta are associated with a significantly larger number of peripheral junctions between endoplasmic reticulum and plasma membrane, which are further enhanced upon stable co-expression of junctate and TRPC3. The gap between the membranes of peripheral junctions is bridged by regularly spaced electron-dense structures of 10 nm. Ins(1,4,5)P3 inhibits the interaction of the cytoplasmic N-terminus of junctate with the ligand-binding domain of the Ins(1,4,5)P3 receptor. Furthermore, Ca2+ influx evoked by activation of Ins(1,4,5)P3 receptors is increased where puncta are located. We conclude that stable peripheral junctions between the plasma membrane and endoplasmic reticulum are the anatomical sites of agonist-activated Ca2+ entry. PMID:21062895
Using dSTORM to probe the molecular architecture of filopodia
NASA Astrophysics Data System (ADS)
Ahmed, Sohail; Chou, Amy; Sem, K. P.; Thankiah, Sudaharan; Wright, Graham; Lim, John; Hariharan, Srivats
2014-03-01
IRSp53 is a Cdc42 effector and a member of the Inverse-Bin-Amphiphysins-Rvs (I-BAR) domain family which can induce negative membrane curvature. IRSp53 generates filopodia by coupling membrane protrusion (I-BAR domain) with actin dynamics through its SH3 domain binding partners. Dynamin 1 (Dyn1), a large GTPase associated with endocytosis, is a novel interacting partner of IRSp53 that localises to filopodia. Using rapid time-lapse TIRF microscopy we have shown that Dyn1 localized to a subcellular region just behind Mena at the leading edge, or in filopodial tip complexes when co-expressed with IRSp53. Dyn1-GFP was strongly localized in the filopodial shaft during the early phase of elongation, after which it moved rearward, suggestive of a role in early filopodia assembly. Mena and Eps8, accumulate at the tip complex in sequence and are involved in filopodial extension and retraction, respectively (Chou at al, 2014 submitted). Here we describe the use of dSTORM to investigate the molecular architecture of filopodia and in particular the size of the F-actin bundle in these structures. The data suggest that direct Stochastic Optical Reconstruction Microscopy (dSTORM) in combination with other techniques will allow the molecular architecture of
A novel role for WAVE1 in controlling actin network growth rate and architecture
Sweeney, Meredith O.; Collins, Agnieszka; Padrick, Shae B.; Goode, Bruce L.
2015-01-01
Branched actin filament networks in cells are assembled through the combined activities of Arp2/3 complex and different WASP/WAVE proteins. Here we used TIRF and electron microscopy to directly compare for the first time the assembly kinetics and architectures of actin filament networks produced by Arp2/3 complex and dimerized VCA regions of WAVE1, WAVE2, or N-WASP. WAVE1 produced strikingly different networks from WAVE2 or N-WASP, which comprised unexpectedly short filaments. Further analysis showed that the WAVE1-specific activity stemmed from an inhibitory effect on filament elongation both in the presence and absence of Arp2/3 complex, which was observed even at low stoichiometries of WAVE1 to actin monomers, precluding an effect from monomer sequestration. Using a series of VCA chimeras, we mapped the elongation inhibitory effects of WAVE1 to its WH2 (“V”) domain. Further, mutating a single conserved lysine residue potently disrupted WAVE1's inhibitory effects. Taken together, our results show that WAVE1 has unique activities independent of Arp2/3 complex that can govern both the growth rates and architectures of actin filament networks. Such activities may underlie previously observed differences between the cellular functions of WAVE1 and WAVE2. PMID:25473116
ERIC Educational Resources Information Center
Smith, Jan
2016-01-01
Research ethics in education is a challenging topic to teach and to learn. As the staff and student body in UK higher education and elsewhere diversifies, the challenges increase as shared reference points diminish. My teaching reflections focus on a key tension explored in this article: how the imperative of internationalising the curriculum…
"Distance Education: An International Journal"-- Reflections on How It All Began
ERIC Educational Resources Information Center
Mitchell, Ian McD.
2009-01-01
In this article, the author reflects on the historical context of the Australian distance education environment that led to the establishment of the journal "Distance Education." He deals with the dynamics of and the decisions made about the journal, in the light of that environment. He also asks readers to share their stance on the…
26 CFR 1.312-1 - Adjustment to earnings and profits reflecting distributions by corporations.
Code of Federal Regulations, 2011 CFR
2011-04-01
... earnings and profits. The reduction in earnings and profits by reason of such distribution is $5,000. Such...,000. The reduction in earnings and profits is $15,000. Such is the reduction even though only the... 26 Internal Revenue 4 2011-04-01 2011-04-01 false Adjustment to earnings and profits reflecting...
Reflections--Dell C. Kjer: A Pioneer Teacher Educator and Association Leader.
ERIC Educational Resources Information Center
Gilstrap, Robert L.
2000-01-01
Reflects upon the role of Dell C. Kjer, a leader in teacher education and president of the Association for Childhood Education International (ACEI) from 1965-1967. Describes his early years in Wisconsin, military duties and student life at George Peabody, his contributions to teacher education, and his contributions to ACEI and other professional…
Reflections on the Laws of Life
ERIC Educational Resources Information Center
Elias, Maurice J.; Klein, Dena A.; DeLuca, Amy; Smith, David R.; Fattal, Laura F.; Bento, Angela; Leverett, Larry
2006-01-01
The Laws of Life is an international essay contest and social and emotional learning (SEL) initiative that many school districts use to build students' SEL and related abilities. In the contest, students reflect on their identities--who they are, who they want to become, and what they want their lives to stand for--by writing about the main values…
1979-04-25
Airport (Bedford, MA ) and Ft. Devens, MA. (2) validation of the models for building reflections based on elevation field measurements at JFK airport and...angles. 2-60 III. BUILDING REFLECTIONS A. Van Measurements at John F. Kennedy (JFK) International Airport, New York Figure 3-1 shows a map of JFK airport with
From Content to Form: Judy Chicago's Pedagogy with Reflections by Judy Chicago
ERIC Educational Resources Information Center
Keifer-Boyd, Karen
2007-01-01
In this article, internationally renowned artist and educator Judy Chicago reflects on her teaching and on my own interpretation of her pedagogy in three projects: "Womanhouse" (1971-1972), "At Home" (2001-2002), and "Envisioning the Future" (2003-2004). From a comparison of pre- and post-open-ended questionnaire responses given by 62 participants…
Audit Cultures, Labour, and Conservative Movements in the Global University
ERIC Educational Resources Information Center
Apple, Michael W.
2013-01-01
I want to use this essay -- basically a commentary -- as a context for some political reflections on what is happening to the governance and the labour processes at universities internationally. In the process, in addition to my critical reflections on the neoliberal impulses affecting universities, I want to do two other things. First, I shall…
From "Borrowing" to "Learning" in International Comparative Study: A Critical Reflection
ERIC Educational Resources Information Center
de Wet, Corene; Wolhuter, Charl
2007-01-01
The state of public education debate, which to a great extent only reflects policies and practices ab road, is examined. The process of learning from others should replace the process of borrowing (of usually inappropriate policies/practices) from others. Two examples of the issues involved in the debate on public education, namely, Outcomes-Based…
ERIC Educational Resources Information Center
Greene, Catie A.
2016-01-01
The purpose of this research was to investigate the sociocultural model of eating disorder development among male and female college student-athletes as moderated by students' level of Reflective Judgment, a stage theory of adult epistemology marked by increasing cognitive complexity. A review of literature on the established relationships between…
Introducing E-Portfolios to Pre-Service Teachers as Tools for Reflection and Growth: Lessons Learnt
ERIC Educational Resources Information Center
Oakley, Grace; Pegrum, Mark; Johnston, Shannon
2014-01-01
E-portfolios are being introduced into initial teacher education programmes internationally to help pre-service teachers build records of their learning journeys and develop into reflective practitioners; to allow them to assemble collections of evidence of their achievements against graduate standards, which can function as digital CVs; and to…
The Value of Reflection in Writing Courses in ELT Preservice Teacher Education Programs
ERIC Educational Resources Information Center
Arikan, Arda
2006-01-01
Constructivist theory has brought significant momentum to all aspects of teacher education. Currently, personal growth of the individual in educational domains is so important that teacher candidates are asked to develop their reflective skills in many courses so that they grow by having internalized and problematized the issues under study by…
Cyclic Colour Change in the Bearded Dragon Pogona vitticeps under Different Photoperiods
Fan, Marie; Stuart-Fox, Devi; Cadena, Viviana
2014-01-01
The ability to change colour rapidly is widespread among ectotherms and has various functions including camouflage, communication and thermoregulation. The process of colour change can occur as an aperiodic event or be rhythmic, induced by cyclic environmental factors or regulated by internal oscillators. Despite the importance of colour change in reptile ecology, few studies have investigated the occurrence of a circadian rhythm in lizard pigmentation. Additionally, although colour change also entails changes in near-infrared reflectance, which may affect thermoregulation, little research has examined this part of the spectrum. We tested whether the bearded dragon lizard, Pogona vitticeps, displays an endogenous circadian rhythm in pigmentation changes that could be entrained by light/dark (LD) cycles and how light affected the relative change in reflectance in both ultraviolet-visible and near-infrared spectra. We subjected 11 lizards to four photoperiodic regimens: LD 12∶12; LD 6∶18; LD 18∶6 and DD; and measured their dorsal skin reflectance at 3-hour intervals for 72 hours after a habituation period. A proportion of lizards displayed a significant rhythm under constant darkness, with maximum reflectance occurring in the subjective night. This endogenous rhythm synchronised to the different artificial LD cycles, with maximum reflectance occurring during dark phases, but did not vary in amplitude. In addition, the total ultraviolet-visible reflectance in relation to the total near-infrared reflectance was significantly higher during dark phases than during light phases. We conclude that P. vitticeps exhibits a circadian pigmentation rhythm of constant amplitude, regulated by internal oscillators and that can be entrained by light/dark cycles. PMID:25354192
Cyclic colour change in the bearded dragon Pogona vitticeps under different photoperiods.
Fan, Marie; Stuart-Fox, Devi; Cadena, Viviana
2014-01-01
The ability to change colour rapidly is widespread among ectotherms and has various functions including camouflage, communication and thermoregulation. The process of colour change can occur as an aperiodic event or be rhythmic, induced by cyclic environmental factors or regulated by internal oscillators. Despite the importance of colour change in reptile ecology, few studies have investigated the occurrence of a circadian rhythm in lizard pigmentation. Additionally, although colour change also entails changes in near-infrared reflectance, which may affect thermoregulation, little research has examined this part of the spectrum. We tested whether the bearded dragon lizard, Pogona vitticeps, displays an endogenous circadian rhythm in pigmentation changes that could be entrained by light/dark (LD) cycles and how light affected the relative change in reflectance in both ultraviolet-visible and near-infrared spectra. We subjected 11 lizards to four photoperiodic regimens: LD 12:12; LD 6:18; LD 18:6 and DD; and measured their dorsal skin reflectance at 3-hour intervals for 72 hours after a habituation period. A proportion of lizards displayed a significant rhythm under constant darkness, with maximum reflectance occurring in the subjective night. This endogenous rhythm synchronised to the different artificial LD cycles, with maximum reflectance occurring during dark phases, but did not vary in amplitude. In addition, the total ultraviolet-visible reflectance in relation to the total near-infrared reflectance was significantly higher during dark phases than during light phases. We conclude that P. vitticeps exhibits a circadian pigmentation rhythm of constant amplitude, regulated by internal oscillators and that can be entrained by light/dark cycles.
Roberts, Kasey; Park, Thomas; Elder, Nancy C; Regan, Saundra; Theodore, Sarah N; Mitchell, Monica J; Johnson, Yolanda N
2015-11-01
Urban Health Project (UHP) is a mission and vision-driven summer internship at the University of Cincinnati College of Medicine that places first-year medical students at local community agencies that work with underserved populations. At the completion of their internship, students write Final Intern Reflections (FIRs). Final Intern Reflections written from 1987 to 2012 were read and coded to both predetermined categories derived from the UHP mission and vision statements and new categories created from the data themselves. Comments relating to UHP's mission and vision were found in 47% and 36% of FIRs, respectively. Positive experiences outweighed negative by a factor of eight. Interns reported the following benefits: educational (53%), valuable (25%), rewarding (25%), new (10%), unique (6%), and life-changing (5%). Urban Health Project is successful in providing medical students with enriching experiences with underserved populations that have the potential to change their understanding of vulnerable populations.
Local in Practice: Professional Distinctions in Angolan Development Work
Peters, Rebecca Warne
2017-01-01
Development workers employed by international nongovernmental organizations (NGOs) are commonly classified as national (local) or international (expatriate) staff members. The distinction is presumed to reflect the varieties of expertise required for the work and the workers’ different biographies. I examine the experiences of Angolans working in an international democratization program to demonstrate how some professionals at the lowest tiers of international development NGOs engage in social practices that strategically emphasize or conceal certain skills, kinds of knowledge, or family circumstances to fulfill industry expectations of “local staff.” Doing so allows them access to employment with international organizations and pursuit of a variety of personal and professional goals. These practices reinforce hierarchical inequalities within the development industry, however, limiting these workers’ influence over programmatic action. I argue that professional distinctions among development workers are social achievements and instruments of strategic manipulation by individuals and NGOs rather than accurate reflections of work or workers. The case study provides insight into the institutional reproduction of hierarchical inequalities and the complexly social reasons why those who suffer their limitations may act in ways that reinforce, rather than resist, unequal social structures. PMID:29430019
Ulvund, Ingeborg; Mordal, Elin
2017-08-01
Offering nursing students' international clinical placement during the educational program is one response to meet the need of cultural competence among nurses. This paper provides insight into the impact of clinical placement, in a developing country, on third year nursing students. In the study we investigated how short term international clinical placement impacted Norwegian nursing students' development of cultural competency. In this study we utilised a qualitative descriptive design and used individual interviews with eighteen Norwegian nursing students who had all participated in an international clinical placement. The data were analysed using the principles of systematic text condensation. In spite the international clinical placement only was four weeks, the findings suggested that real life experience culturally awakened the students and forced an ongoing process developing cultural competence. However, it is important to give students time to reflection. Although increased cultural awareness and a growing cultural competence was identified by the students undertaking international clinical placement, further research is required. It is important to investigate the best methods to support the students' reflection such that the experiences lead to learning. Copyright © 2017 Elsevier Ltd. All rights reserved.
Personal Growth During Internship: A Qualitative Analysis of Interns' Responses to Key Questions
Levine, Rachel B; Haidet, Paul; Kern, David E; Beasley, Brent W; Bensinger, Lisa; Brady, Donald W; Gress, Todd; Hughes, Jennifer; Marwaha, Ajay; Nelson, Jennifer; Wright, Scott M
2006-01-01
BACKGROUND During clinical training, house officers frequently encounter intense experiences that may affect their personal growth. The purpose of this study was to explore processes related to personal growth during internship. DESIGN Prospective qualitative study conducted over the course of internship. PARTICIPANTS Thirty-two postgraduate year (PGY)-1 residents from 9 U.S. internal medicine training programs. APPROACH Every 8 weeks, interns responded by e-mail to an open-ended question related to personal growth. Content analysis methods were used to analyze the interns' writings to identify triggers, facilitators, and barriers related to personal growth. RESULTS Triggers for personal growth included caring for critically ill or dying patients, receiving feedback, witnessing unprofessional behavior, experiencing personal problems, and dealing with the increased responsibility of internship. Facilitators of personal growth included supportive relationships, reflection, and commitment to core values. Fatigue, lack of personal time, and overwhelming work were barriers to personal growth. The balance between facilitators and barriers may dictate the extent to which personal growth occurs. CONCLUSIONS Efforts to support personal growth during residency training include fostering supportive relationships, encouraging reflection, and recognizing interns' core values especially in association with powerful triggers. PMID:16808737
Nishigori, Hiroshi; Otani, Takashi; Plint, Simon; Uchino, Minako; Ban, Nobutaro
2009-05-01
Although medical students have increasingly more opportunities to participate in international electives, their experiences are usually unstructured and the literature referring to their learning outcomes, educational environment, and assessment is scanty. This study was undertaken to clarify qualitatively what students learn from their international electives. We carried out semi-structured individual interviews with 15 Japanese students studying clinical medicine in British medical schools and six British students studying in Japanese medical schools. The thematic synthesis method was used in analysing the transcribed data and triangulation by multiple researchers was used to achieve higher reliability. The main learning outcomes identified were skills in history taking and physical examination with clinical reasoning and in management of diseases rarely seen in the students' own countries; awareness of clinical ethics and merits and demerits of different systems of healthcare and medical education; sensitivity to issues in doctor-patient relationships and work ethics; enhancement of cultural competence; and personal development. Most learning outcomes of international electives are culture- or system-dependent. Students achieved outcomes related closely to medical professionalism, mainly through reflection. International electives may give students opportunities to learn both professionalism and cultural competence.
Memory: Enduring Traces of Perceptual and Reflective Attention
Chun, Marvin M.; Johnson, Marcia K.
2011-01-01
Attention and memory are typically studied as separate topics, but they are highly intertwined. Here we discuss the relation between memory and two fundamental types of attention: perceptual and reflective. Memory is the persisting consequence of cognitive activities initiated by and/or focused on external information from the environment (perceptual attention) and initiated by and/or focused on internal mental representations (reflective attention). We consider three key questions for advancing a cognitive neuroscience of attention and memory: To what extent do perception and reflection share representational areas? To what extent are the control processes that select, maintain, and manipulate perceptual and reflective information subserved by common areas and networks? During perception and reflection, to what extent are common areas responsible for binding features together to create complex, episodic memories and for reviving them later? Considering similarities and differences in perceptual and reflective attention helps integrate a broad range of findings and raises important unresolved issues. PMID:22099456
Minter, Rebecca M; Amos, Keith D; Bentz, Michael L; Blair, Patrice Gabler; Brandt, Christopher; D'Cunha, Jonathan; Davis, Elisabeth; Delman, Keith A; Deutsch, Ellen S; Divino, Celia; Kingsley, Darra; Klingensmith, Mary; Meterissian, Sarkis; Sachdeva, Ajit K; Terhune, Kyla; Termuhlen, Paula M; Mullan, Patricia B
2015-08-01
To evaluate interns' perceived preparedness for defined surgical residency responsibilities and to determine whether fourth-year medical school (M4) preparatory courses ("bootcamps") facilitate transition to internship. The authors conducted a multi-institutional, mixed-methods study (June 2009) evaluating interns from 11 U.S. and Canadian surgery residency programs. Interns completed structured surveys and answered open-ended reflective questions about their preparedness for their surgery internship. Analyses include t tests comparing ratings of interns who had and had not participated in formal internship preparation programs. The authors calculated Cohen d for effect size and used grounded theory to identify themes in the interns' reflections. Of 221 eligible interns, 158 (71.5%) participated. Interns self-reported only moderate preparation for most defined care responsibilities in the medical knowledge and patient care domains but, overall, felt well prepared in the professionalism, interpersonal communication, practice-based learning, and systems-based practice domains. Interns who participated in M4 preparatory curricula had higher self-assessed ratings of surgical technical skills, professionalism, interpersonal communication skills, and overall preparation, at statistically significant levels (P < .05) with medium effect sizes. Themes identified in interns' characterizations of their greatest internship challenges included anxiety or lack of preparation related to performance of technical skills or procedures, managing simultaneous demands, being first responders for critically ill patients, clinical management of predictable postoperative conditions, and difficult communications. Entering surgical residency, interns report not feeling prepared to fulfill common clinical and professional responsibilities. As M4 curricula may enhance preparation, programs facilitating transition to residency should be developed and evaluated.
A comprehensive ray tracing study on the impact of solar reflections from glass curtain walls.
Wong, Justin S J
2016-01-01
To facilitate the investigation of the impact of solar reflection from the façades of skyscrapers to surrounding environment, a comprehensive ray tracing model has been developed using the International Commerce Centre (ICC) in Hong Kong as an example. Taking into account the actual physical dimensions of buildings and meteorological data, the model simulates and traces the paths of solar reflections from ICC to the surrounding buildings, assessing the impact in terms of hit locations, light intensity and the hit time on each day throughout the year. Our analyses show that various design and architectural features of ICC have amplified the intensity of reflected solar rays and increased the hit rates of surrounding buildings. These factors include the high reflectivity of glass panels, their upward tilting angles, the concave profile of the 'Dragon Tail' (glass panels near the base), the particular location and orientation of ICC, as well as the immense height of ICC with its large reflective surfaces. The simulation results allow us to accurately map the date and time when the ray projections occur on each of the target buildings, rendering important information such as the number of converging (overlapping) projections, and the actual light intensity hitting each of the buildings at any given time. Comparisons with other skyscrapers such as Taipei 101 in Taiwan and 2-IFC (International Finance Centre) Hong Kong are made. Remedial actions for ICC and preventive measures are also discussed.
Developing a Value Framework: The Need to Reflect the Opportunity Costs of Funding Decisions.
Sculpher, Mark; Claxton, Karl; Pearson, Steven D
2017-02-01
A growing number of health care systems internationally use formal economic evaluation methods to support health care funding decisions. Recently, a range of organizations have been advocating forms of analysis that have been termed "value frameworks." There has also been a push for analytical methods to reflect a fuller range of benefits of interventions through multicriteria decision analysis. A key principle that is invariably neglected in current and proposed frameworks is the need to reflect evidence on the opportunity costs that health systems face when making funding decisions. The mechanisms by which opportunity costs are realized vary depending on the system's financial arrangements, but they always mean that a decision to fund a specific intervention for a particular patient group has the potential to impose costs on others in terms of forgone benefits. These opportunity costs are rarely explicitly reflected in analysis to support decisions, but recent developments to quantify benefits forgone make more appropriate analyses feasible. Opportunity costs also need to be reflected in decisions if a broader range of attributes of benefit is considered, and opportunity costs are a key consideration in determining the appropriate level of total expenditure in a system. The principles by which opportunity costs can be reflected in analysis are illustrated in this article by using the example of the proposed methods for value-based pricing in the United Kingdom. Copyright © 2017 International Society for Pharmacoeconomics and Outcomes Research (ISPOR). Published by Elsevier Inc. All rights reserved.
Behrens, Johann
2010-01-01
Evidence-based Medicine (EbM) is the ongoing self-reflection of an individualised approach to medicine in terms of a science that originates from and focuses on clinical decision-making (pragmatic science="Handlungswissenschaft"). EbM is particularly suitable for self-reflecting individualised medicine on the basis of decision-oriented pragmatic science because it consistently distinguishes between external evidence (i.e., other subjects' experience gained through "qualitative" and "quantitative" scientific methods) and internal evidence, i.e., the individual user's, or patient's, own experience manifesting and developing in the individual contact between therapist and patient. Therefore, internal evidence is completely different from the individual clinical experience, expertise, and conviction which therapists contribute to the encounter with clients. A deeper understanding of internal evidence as a result of this encounter has emerged only in the past 15 years. However, it is an integral part of the logic of evidence-based professional decision-making. Scientifically justified beneficial and effective treatment in the individual case cannot be deduced from external evidence but can only be gathered from internal evidence for which the best external evidence available has been utilised. In the past 15 years nursing science has not only carved out the decision-oriented scientific core of evidence-based practice but has also tried to increase the validity of studies on external evidence by employing a combination of 'qualitative' social science studies and clinical epidemiological methods. Copyright © 2010. Published by Elsevier GmbH.
Research advances in reflectance spectra of plant leafs
NASA Astrophysics Data System (ADS)
Zhu, Taotao; Yang, Ting; Guo, Yanxin; Xu, Jingqi; Chang, Wandong; Fang, Siyi; Zhu, Kangkang; Xu, Tingyan
2018-02-01
Leaves are an important factor when we study plants because their water content, pigment content and nutrient content of leaves can reflect the current growth status of the whole plant. The methods of spectral diagnosis technology or image technology mainly are the pre-detection technique which can be used to invert the color, texture and spectral reflectance of the leaves. From this we can obtain the changes of the internal components and the external morphological characteristics of the plant leaves in different states changes. In this paper, the reflection spectral response mechanism of plant water content, pigment and nutrient elements at domestic and overseas are reviewed and compared.
Micro ring cavity resonator incorporating total internal reflection mirrors
NASA Astrophysics Data System (ADS)
Kim, Doo Gun; Choi, Woon Kyung; Choi, Young Wan; Yi, Jong Chang; Chung, Youngchul; Dagli, Nadir
2007-02-01
We investigate the properties of a multimode-interference (MMI) coupled micro ring cavity resonator with total-internal-reflection (TIR) mirrors and a semiconductor optical amplifier (SOA). The TIR mirrors were fabricated by the self-aligned process with a loss of 0.7 dB per mirror. The length and width of an MMI are 142 μm and 10 μm, respectively. The resulting free spectral range (FSR) of the resonator was approximately 1.698 nm near 1571 nm and the extinction ratio was about 17 dB. These devices might be useful as optical switching and add-drop filters in a photonic integrated circuit or as small and fast resonator devices.
Chapter 7: Total internal reflection fluorescence microscopy.
Axelrod, Daniel
2008-01-01
Total internal reflection fluorescence microscopy (TIRFM), also known as evanescent wave microscopy, is used in a wide range of applications, particularly to view single molecules attached to planar surfaces and to study the position and dynamics of molecules and organelles in living culture cells near the contact regions with the glass coverslip. TIRFM selectively illuminates fluorophores only in a very thin (less than 100 nm deep) layer near the substrate, thereby avoiding excitation of fluorophores outside this subresolution optical section. This chapter reviews the history, current applications in cell biology and biochemistry, basic optical theory, combinations with numerous other optical and spectroscopic approaches, and a range of setup methods, both commercial and custom.
NASA Astrophysics Data System (ADS)
Sung, Young Hoon; Jung, Pil-Hoon; Han, Kyung-Hoon; Kim, Yang Doo; Kim, Jang-Joo; Lee, Heon
2017-10-01
In order to increase the out-coupling efficiency of organic light emitting diodes, conical Si oxide nanostructures were formed on a glass substrate using nanoimprint lithography with hydrogen silsesquioxane. Then, the substrate was planarized with TiO2 nanoparticles. Since TiO2 nanoparticles have a higher refractive index than Si oxide, the surface of substrate is physically flat, but optically undulated in a manner that enables optical scattering and suppression of total internal reflection. Subsequently, OLEDs formed on a substrate with nanostructured Si oxide and a TiO2 planarization layer exhibit a 25% increase in out-coupling efficiency by suppressing total internal reflection.
Road to Ahmedabad: Embedding Environmental Wisdom in Our Cultural DNA
ERIC Educational Resources Information Center
Hopkins, Charles
2009-01-01
In his speech at the inaugural session of the 4th International Conference on Environmental Education, Ahmedabad, India, in November 2007, Charles Hopkins, one of the "founding fathers" of environmental education (EE) at the 1st International Conference on Environmental Education in Tbilisi, Georgia, in 1977, reflected on how EE has…
The Challenges to and the Need for International Research in Educational Leadership
ERIC Educational Resources Information Center
Brooks, Melanie Carol; Jean-Marie, Gaetane
2015-01-01
Purpose: The purpose of this paper is twofold: to discuss methodological challenges facing US scholars when conducting international research; and to present personal reflections as educational leadership faculty in the USA conducting and publishing on research undertaken in Haiti and Thailand. Design/Methodology/Approach: This study drew from…
International Reports on Literacy Research: Chile, Argentina, Brazil, and Colombia
ERIC Educational Resources Information Center
Malloy, Jacquelynn A., Comp.; Botzakis, Stergios, Comp.
2005-01-01
This is a compilation of reports on international literacy research. The report includes 4 separate reports on Chile, Argentina, Brazil, and Colombia. In the first report, research correspondent Marta Infante reports on two studies that reflect the growing interest of Chilean professionals in studying reading-related factors such as phonemic…
ERIC Educational Resources Information Center
Rahatzad, Jubin; Sasser, Hannah L.; Phillion, JoAnn; Karimi, Nastaran; Deng, Yuwen; Akiyama, Reiko; Sharma, Suniti
2013-01-01
Preservice teachers' international cross-cultural experiences can provide opportunities for the exploration of epistemic frontiers. In this article we suggest that postglobal teacher preparation take a critically reflective approach that engages preservice teachers in border thinking, which allows for other ways of knowing while studying abroad.…
International Children's Trade Books: Building Blocks for Character Education
ERIC Educational Resources Information Center
Young, Terrell A.; Hadaway, Nancy L.; Ward, Barbara A.
2013-01-01
The importance of character education has been emphasized since ancient times across cultures and religions as a way to develop morals and values of the younger generations. In this article, the authors highlight several award-winning international children's trade books that reflect the core values recurring in literature and scriptures…
Reflections on Dead Theory in International Relations
ERIC Educational Resources Information Center
Thakur, Vineet
2016-01-01
In this short autobiographical essay, I trace my journey in the discipline of International Relations. While entering the discipline, I, along with a host of my classmates, were enamoured by the exciting possibilities of thinking theoretically. Almost a decade later, those promises look bleak. From the perspective of a student in the discipline, I…
ERIC Educational Resources Information Center
Savvides, Nicola; Al-Youssef, Joanna; Colin, Mindy; Garrido, Cecilia
2014-01-01
This article highlights key theoretical and methodological issues and implications of being an insider/outsider when undertaking qualitative research in international educational settings. It first addresses discourses of "self" and "other," noting that identity and belonging emerge from fluid engagement between researchers and…
Pestalozzi: Foster Father of Early Childhood Education.
ERIC Educational Resources Information Center
Hewes, Dorothy W.
In tracing the spread of the educational philosophy of Johann Heinrich Pestalozzi, it is useful to understand educators' emphasis on an internal or external locus of control. Pestalozzi was an individual with an internal locus of control, and this trait was reflected in his educational philosophy of self-learning and free investigation. However,…
Student Selection for Higher Education: The Relationship Between Internal and External Marks.
ERIC Educational Resources Information Center
Papas, George; Psacharopoulos, George
1993-01-01
Longitudinal data on 366 Greek higher education candidates are used to explore relationship between internal (school average) and external (entrance examinations) marks. Marks of last three grades of secondary school seem to reflect fairly well how student will perform in external examinations. Questions of access to education and tests as…
iiESI Wraps Up First International Conference | News | NREL
. Over the course of the event, presentations touched on sub-topics of the conference theme, " topics and research interests resulted in a truly useful and productive exercise," said O'Malley . Such a range in topics was reflective of the institutions that compose iiESI ---- an international
Highlights from the First World Congress of Education International.
ERIC Educational Resources Information Center
Education International, Brussels (Belgium).
The papers in this volume reflect the general theme of the congress, "Educators United, Ready for Change." A theme that emerged during the conference concerned the need to fight structural adjustment programs, imposed by the World Bank or the International Monetary Fund, which were destroying the educational and social infrastructure of…
Reflecting on the Role of Competence and Culture in Consultation at the International Level
ERIC Educational Resources Information Center
Rosenfield, Sylvia
2014-01-01
International educational consultation is challenging work that requires not only attention to best practices in consultation but also additional focus on cultural norms and regularities. In the three articles of this special issue, the consultation competencies of consultants play a critical role, as exemplified by entry issues, problem-solving…
International Videoconferencing for Public Health Education: Linking the U.S. and Germany
ERIC Educational Resources Information Center
Burke, Sloane C.; Chaney, Beth H.; Kirsten, Wolf
2010-01-01
Background: Video-linking with other countries provides a low-cost vehicle for students to learn more about health issues of other cultures and reflectively discuss the public health promotion and education activities of those countries in real time. Purpose: This article presents a case study of international videoconferencing in the higher…
Participation and Service Access Rights for People with Intellectual Disability: A Role for Law?
ERIC Educational Resources Information Center
Carney, Terry
2013-01-01
Background: Supported decision-making and personal budgets for services are the new paradigms. Method: Supported decision-making proposals from the Australian State of Victoria are analysed against international trends to determine the viability of laws reflecting new international norms of the United Nations Convention on the Rights of Persons…
ERIC Educational Resources Information Center
Sun, Qi; Roumell, Elizabeth A. L.
2017-01-01
Applying international and comparative education and cultural perspectives, through critical comparative reflection, this paper examines issues and challenges of adult learning, teaching, and research endeavors between the East and the West. First, a brief literature review illuminating the obstacles and barriers for a two-way exchange between…
ERIC Educational Resources Information Center
Shah, Ritesh; McCormick, Alexander; Thomas, Matthew A. M.
2017-01-01
In this paper, we critically interrogate the way in which comparative and international education coursework at two large institutions in Australia and New Zealand embody or challenge teleological, colonial, and Western/Northern-centric perspectives on education and development. Embedded within a broader and introspective examination of our roles…
ERIC Educational Resources Information Center
Benzie, Helen Joy
2010-01-01
The current concern about low levels of English proficiency among international students who graduate from degree courses--that students' English language skills are not being developed during their higher education experience--reflects negatively on the quality of Australian higher education and its graduates. More careful selection of students…
ASBO at 100: A Century of School Business Management
ERIC Educational Resources Information Center
School Business Affairs, 2010
2010-01-01
This article talks about Association of School Business Officials International (ASBO) at a century of school business management. As ASBO International celebrates its centennial, one should reflect on the truth of this often-heard quote and how it applies to the field of school business management in general and the association in particular.…
Promising Practices to Meet Global Challenges in Science and Mathematics Education
ERIC Educational Resources Information Center
Berlin, Donna F., Ed.; White, Arthur L., Ed.
2010-01-01
The chapters in this book reflect the work of science and mathematics educators who have worked for many years at the international level. As members of the International Consortium for Research in Science and Mathematics Education, their work provides readers with issues, models, practices, and research results that have applicability and…
Decision Making for Democratic Leadership in a Guided Internship
ERIC Educational Resources Information Center
Klinker, JoAnn Franklin; Hoover, J. Duane; Valle, Fernando; Hardin, Fred
2014-01-01
Experience in problem-based learning, authentic experiences, on-the-job decision making, and critical reflection on decisions made formed the conceptual framework of an internship to develop democratic leadership as a professional ethic in interns. Interns in an on-the-job guided internship examined decisions over a 13-week period as they…
ERIC Educational Resources Information Center
Wright, J.W., Jr.
2010-01-01
This publication is intended for three primary audiences: international development professionals who want to define "workforce" for program design purposes, those who seek information about field activities, and those who want to reflect on the implementation of successful, high impact programs--whether they were broad-based or…
Examining External and Internal Poverty as Antecedents of Teen Pregnancy
ERIC Educational Resources Information Center
Young, Tamera; Turner, Jean; Denny, George; Young, Michael
2004-01-01
Objectives: To identify antecedents of teen pregnancy. Methods: Data from the National Education Longitudinal Study were analyzed. This data set allowed us to identify eighth-grade antecedents of teen pregnancy/childbearing. Results: The variables that were found to be most predictive of later pregnancy were reflective of internal poverty (locus…
ERIC Educational Resources Information Center
Gribble, Nigel; Dender, Alma; Lawrence, Emma; Manning, Kirrily; Falkmer, Torbjorn
2014-01-01
In the increasingly global world, skills in cultural competence now form part of the minimum standards of practice required for allied health professionals. During an international work-integrated learning (WIL) placement, allied health students' cultural competence is expected to be enhanced. The present study scrutinized reflective journals of…
International Education in Japan: Response of the Grass-Eaters
ERIC Educational Resources Information Center
Carpenter, Walter
2012-01-01
This paper reviews some of the reasons why a majority of Japanese university students are not interested in study-abroad, international-education type programs. A general consensus in Japan is that disinterest in studying abroad is reflective of the values held by many Japanese young people, often referred to as the "grass-eating"…
ERIC Educational Resources Information Center
Patterson, Timothy
2015-01-01
Common sense thinking on international professional development suggests that the rewards for teachers are automatic. One of the most frequently advertised gains teachers are expected to see from participation includes the likelihood that they will have a transformative experience, whereby aspects of their personal or professional attributes are…
ERIC Educational Resources Information Center
Li, Jiaqi; Wang, Yanlin; Liu, Xun; Xu, Yusu; Cui, Tingting
2018-01-01
This study used a consensual qualitative research method to explore academic adaptation experiences of international students (N = 13) from East Asian countries at a U.S. university. The analysis yielded five domains from the data (challenges, feelings, strategies, suggestions, and self-reflections). Implications for college counselors, university…
Books and National Development. Seminar Report April 27-29, 1968, Academy House, Seoul.
ERIC Educational Resources Information Center
Korean Publishers Association, Seoul.
Representatives from Korea, China, Indonesia, Japan, Thailand, the United States and Southeast Asian Ministers of Education (SEAMES) attended an international seminar to reaffirm the importance of books as national development tools, to seek measures for having it reflected in the national policies and to promote international cooperation in book…
Participant Perspectives and Critical Reflections on Language Teacher Education by Distance
ERIC Educational Resources Information Center
Knox, John S.
2017-01-01
Language teaching is a profession which is international in character. Language teachers often work and study in foreign countries, and distance education has become very important in the education of language teachers. Drawing on two international surveys, this paper explores language teacher education by distance from the perspective of students…
ERIC Educational Resources Information Center
Stoit, Astrid M. B.; van Schie, Hein T.; Riem, Madelon; Meulenbroek, Ruud G. J.; Newman-Norlund, Roger D.; Slaats-Willemse, Dorine I. E.; Bekkering, Harold; Buitelaar, Jan K.
2011-01-01
Qualitative differences in social interaction and communication are diagnostic hallmarks in autism spectrum disorders (ASD). The present study investigated the hypothesis that impaired social interaction in ASD reflects a deficit to internally model the behavior of a co-actor. Children and adolescents with ASD and matched controls performed a…
Blended Learning from Design to Evaluation: International Case Studies of Evidence-Based Practice
ERIC Educational Resources Information Center
Vaughan, Norman; Reali, Aline; Stenbom, Stefan; Van Vuuren, Marieta Jansen; MacDonald, David
2017-01-01
This study compares and contrasts four international faculty development programs for blended learning in order to understand the benefits, challenges, lessons learned, and recommendations from such initiatives. The benefits identified for faculty members, who participated in these programs, were that they became more reflective of their teaching…
ERIC Educational Resources Information Center
Motley, Phillip; Sturgill, Amanda
2013-01-01
This project assessed how an international service-learning course affected mass communication students' knowledge of professionalism. Using written reflections and focus group transcripts from four courses that took place in Central America, we observed that placing students in immersive environments, where they are able to work on authentic…
Strait of Gibraltar as seen from STS-58
1993-10-20
STS058-73-009 (18 Oct-1 Nov 1993) --- Atlantic water flowing with the tide through the Strait of Gibraltar into the Mediterranean generates internal waves as depicted in this photo. The incoming cool, less dense Atlantic water flows over the warm, more saline Mediterranean water. As the tide moves into the Strait of Gibraltar it encounters the Camarinal Sill, which is like a cliff under water, south of Camarinal Point, Spain. Internal waves are generated at the Sill and travel along the density boundary between the Atlantic water and the Mediterranean water. Internal waves have very little effect on the sea surface, except for gentle slopes and slight differences in roughness. We can see them in the Space Shuttle photos because of sunglint which reflects off the water. Internal waves smooth out some of the capillary waves at the surface in bands. The sun reflects more brightly from these smooth areas showing us the pattern of the underwater waves. The Bay of Cadiz on the southwest coast of Spain, the Rock of Gibraltar, and the Moroccan coast are also visible in this photo.
Borelli, Jessica L; Palmer, Alexandra; Vanwoerden, Salome; Sharp, Carla
2017-12-13
Although convergence in parent-youth reports of adolescent psychopathology is critical for treatment planning, research documents a pervasive lack of agreement in ratings of adolescents' symptoms. Attachment insecurity (particularly disorganized attachment) and impoverished reflective functioning (RF) are 2 theoretically implicated predictors of low convergence that have not been examined in the literature. In a cross-sectional investigation of adolescents receiving inpatient psychiatric treatment, we examined whether disorganized attachment and low (adolescent and parent) RF were associated with patterns of convergence in adolescent internalizing and externalizing symptoms. Compared with organized adolescents, disorganized adolescents had lower parent-youth convergence in reports of their internalizing symptoms and higher convergence in reports of their externalizing symptoms; low adolescent self-focused RF was associated with low convergence in parent-adolescent reports of internalizing symptoms, whereas low adolescent global RF was associated with high convergence in parent-adolescent reports of externalizing symptoms. Among adolescents receiving inpatient psychiatric treatment, disorganized attachment and lower RF were associated with weaker internalizing symptom convergence and greater externalizing symptom convergence, which if replicated, could inform assessment strategies and treatment planning in this setting.
Doing Peer Review: Reflections from an International Group of Postdoctoral Fellows
ERIC Educational Resources Information Center
Merry, Lisa; Jarvis, Kimberly; Kupoluyi, Joseph; Lual, Jomama One Jomama
2017-01-01
There is very little written regarding developing the skills of doing peer reviews. In this piece we use our own experience as postdoctoral fellows to offer our reflections on how to get the most out of doing peer reviews as a trainee researcher. We touch upon the variety and complexity of peer reviews, the debates concerning the nature and…
ERIC Educational Resources Information Center
Herrera, Tony Isaac
2010-01-01
This qualitative case study was designed to explore whether and how a sample of domestic and international managers use two key adult education concepts--critical reflection and experiential learning--to influence changes in individual employees whom they coach. The study is based on the primary assumption that although managers do not…
ERIC Educational Resources Information Center
Gogolin, Ingrid
2011-01-01
The review of TLRP's research that is presented in this volume offers manifold aspects that are worth reflecting on. In my contribution, I concentrate on two aspects that are especially interesting from an international perspective or, more precisely, from a German point of view. The first is a general reflection on the education and science…
ERIC Educational Resources Information Center
Gowindasamy, Maniyarasi
2017-01-01
This study was conducted to evaluate the implementation of reflective development model in improving the intercultural competence among business student in Stamford College. This study will be focus on the local and international students in terms of their cultural competencies through the globalization subjects. An embedded design of mixed…
ERIC Educational Resources Information Center
Fierke, Kerry K.; Lepp, Gardner A.
2015-01-01
The article shares the outcomes of a practice called Intention/Reflection (I/R) when applied to a group of ten students in a five-week course involving an international advanced pharmacy practice experience. Developed by the authors and founded on a combination of theoretical principles, this practice is unique because of the blend of formative…
Reflecting on Talk: A Mentor Teacher's Gradual Release in Co-Planning
ERIC Educational Resources Information Center
Pylman, Stacey
2016-01-01
The goal of this case study was to explore how a mentor teacher used video-recorded co-planning sessions to reflect on and improve one's mentoring practice. Findings reveal ways in which the mentor used talk in co-planning sessions to model one's thinking process and to gradually release planning responsibility to engage the intern in learning to…
New light-shielding technique for shortening the baffle length of a star sensor
NASA Astrophysics Data System (ADS)
Kawano, Hiroyuki; Sato, Yukio; Mitani, Kenji; Kanai, Hiroshi; Hama, Kazumori
2002-10-01
We have developed a star sensor with a short baffle of 140 mm. Our baffle provides a Sun rejection angle of 35 degrees with stray light attenuation less than the intensity level of a visual magnitude of Mv = +5 for a wide field of view lens of 13x13 degrees. The application of a new light shielding technique taking advantage of total internal reflection phenomena enables us to reduce the baffle length to about three fourths that of the conventional two-stage baffle. We have introduced two ideas to make the baffle length shorter. The one is the application of a nearly half sphere convex lens as the first focusing lens. The bottom surface reflects the scattering rays with high incident angles of over 50 degrees by using the total internal reflection phenomena. The other is the painting of the surface of the baffle with not frosted but gloss black paint. The gloss black paint enables most of the specular reflection rays to go back to outer space without scattering. We confirm the baffle performance mentioned above by scattering ray tracing simulation and a light attenuation experiment in a darkroom on the ground.
Carter, Amanda G; Creedy, Debra K; Sidebotham, Mary
2017-11-01
develop and test a tool designed for use by academics to evaluate pre-registration midwifery students' critical thinking skills in reflective writing. a descriptive cohort design was used. a random sample (n = 100) of archived student reflective writings based on a clinical event or experience during 2014 and 2015. a staged model for tool development was used to develop a fifteen item scale involving item generation; mapping of draft items to critical thinking concepts and expert review to test content validity; inter-rater reliability testing; pilot testing of the tool on 100 reflective writings; and psychometric testing. Item scores were analysed for mean, range and standard deviation. Internal reliability, content and construct validity were assessed. expert review of the tool revealed a high content validity index score of 0.98. Using two independent raters to establish inter-rater reliability, good absolute agreement of 72% was achieved with a Kappa coefficient K = 0.43 (p<0.0001). Construct validity via exploratory factor analysis revealed three factors: analyses context, reasoned inquiry, and self-evaluation. The mean total score for the tool was 50.48 (SD = 12.86). Total and subscale scores correlated significantly. The scale achieved good internal reliability with a Cronbach's alpha coefficient of .93. this study establishedthe reliability and validity of the CACTiM (reflection) for use by academics to evaluate midwifery students' critical thinking in reflective writing. Validation with large diverse samples is warranted. reflective practice is a key learning and teaching strategy in undergraduate Bachelor of Midwifery programmes and essential for safe, competent practice. There is the potential to enhance critical thinking development by assessingreflective writing with the CACTiM (reflection) tool to provide formative and summative feedback to students and inform teaching strategies. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.
Health Monitoring of Thermal Barrier Coatings by Mid-Infrared Reflectance
NASA Technical Reports Server (NTRS)
Eldridge, J. I.; Spuckler, C. M.; Nesbitt, J. A.; Street, K. W.
2002-01-01
Mid-infrared (MIR) reflectance is shown to be a powerful tool for monitoring the integrity of 8wt% yttria-stabilized zirconia (8YSZ) thermal barrier coatings (TBCs). Because of the translucent nature of plasma-sprayed 8YSZ TBCs, particularly at MIR wavelengths (3 to 5 microns), measured reflectance does not only originate from the TBC surface, but contains strong contributions from internal scattering within the coating as well as reflectance from the underlying TBC/substrate interface. Therefore, changes in MIR reflectance measurements can be used to monitor the progression of TBC delamination. In particular, MIR reflectance is shown to reproducibly track the progression of TBC delamination produced by repeated thermal cycling (to 1163 C) of plasma-sprayed 8YSZ TBCs on Rene N5 superalloy substrates. To understand the changes in MIR reflectance with the progression of a delamination crack network, a four-flux scattering model is used to predict the increase in MIR reflectance produced by the introduction of these cracks.
Lindblad, Åsa Kettis; Gustavsson, Maria; Ring, Lena
2009-01-01
Objective To identify individual and social factors associated with pharmacy students' level of reflection in an advanced pharmacy practice experience (APPE). Methods A postal questionnaire, including a reflective assignment, was sent to all pharmacy interns (n=262) at Uppsala University, Sweden, for 4 semesters in 2005-2007. Results In a univariate analysis, 7 factors were found to be associated with students' level of reflection. After controlling for covariates, 3 social factors were found to be independently associated with reflection: having a formal preceptor (OR=5.3), being at a small pharmacy (OR=19.8), and students' perception of the importance of discussing critical thinking with the preceptor (OR=1.2). No correlation could be observed between level of reflection and critical thinking, nor learning style. Conclusion Social components seem to be of higher importance than individual components in students' reflective levels after pharmacy internship experience. Trained preceptors are important to foster reflection skills. PMID:19885076
NASA Astrophysics Data System (ADS)
Fujinami, Taku; Kigami, Hiroshi; Unno, Noriyuki; Taniguchi, Jun; Satake, Shin-ichi
2018-06-01
Total internal reflection fluorescence microscopy (TIRFM) is a promising method for measuring fluid flow close to a wall with nanoscale resolution in a process that is termed "multilayer nanoparticle image velocimetry" (MnPIV). TIRFM uses evanescent light that is generated on a substrate (typically a glass slide) by total internal reflection of light. Many researchers have previously studied x- y- z (3D) flows of water close to flat glass slides using MnPIV. On the other hand, a fluid flow close to a structured surface is also important. To measure flows of water near micro-patterns, we previously developed an MnPIV technique that uses a refractive-index-matching method. In previous study, the micropattern is made of a thermoplastic material with a refractive index that closely matches that of water. In this study, ultraviolet nanoimprint lithography was used for fabricating the appropriate micro-patterns because this technique can fabricate a pattern with a high resolution. As a result, we succeeded in performing MnPIV in water with a circular hole array pattern made by ultraviolet nanoimprint using a refractive-index-matching method. We believe that this technique will be helpful in elucidating fluid flows around microstructures.
NASA Technical Reports Server (NTRS)
Curley, Michael J. (Inventor); Sarkisov, Sergey S. (Inventor)
2008-01-01
A refractometer computer controls the rotation of a rotary plate upon which are mounted a prism optically coupled via an optical window to a spectroscopic cell holding a resin exhibiting a dynamic refractive index during photocuring. The computer system positions the prism and spectroscopic cell relative to a visible light laser which illuminates the prism-resin interface at selected incidence angles. A photodetector mounted on the plate generates a signal to the computer proportional to intensity of an internally reflected light beam. A curing light is selectively transmitted through the prism and into the photocurable resin. The refractometer determines the intensity of the internally reflected beam a selected incidence angles and determines the effective refractive index curve of the resin at an uncured state and, optionally, at a completely cured state. Next, an amount of uncured resin and selected optical components to be joined by the resin is placed in the spectroscopic cell and irradiated with the UV light. The refractometer is fixed at a selected incidence angle and measures the intensity of an internally reflected light beam of light throughout the cure cycle. The refractometer determines the resin's refractive index of the polymeric mixture by means of extrapolation of a horizontal shift in the effective refractive index curve of the resin from an uncured state to a selected point in the cure cycle.
Berendt, Mette; Farquhar, Robyn G; Mandigers, Paul J J; Pakozdy, Akos; Bhatti, Sofie F M; De Risio, Luisa; Fischer, Andrea; Long, Sam; Matiasek, Kaspar; Muñana, Karen; Patterson, Edward E; Penderis, Jacques; Platt, Simon; Podell, Michael; Potschka, Heidrun; Pumarola, Martí Batlle; Rusbridge, Clare; Stein, Veronika M; Tipold, Andrea; Volk, Holger A
2015-08-28
Dogs with epilepsy are among the commonest neurological patients in veterinary practice and therefore have historically attracted much attention with regard to definitions, clinical approach and management. A number of classification proposals for canine epilepsy have been published during the years reflecting always in parts the current proposals coming from the human epilepsy organisation the International League Against Epilepsy (ILAE). It has however not been possible to gain agreed consensus, "a common language", for the classification and terminology used between veterinary and human neurologists and neuroscientists, practitioners, neuropharmacologists and neuropathologists. This has led to an unfortunate situation where different veterinary publications and textbook chapters on epilepsy merely reflect individual author preferences with respect to terminology, which can be confusing to the readers and influence the definition and diagnosis of epilepsy in first line practice and research studies.In this document the International Veterinary Epilepsy Task Force (IVETF) discusses current understanding of canine epilepsy and presents our 2015 proposal for terminology and classification of epilepsy and epileptic seizures. We propose a classification system which reflects new thoughts from the human ILAE but also roots in former well accepted terminology. We think that this classification system can be used by all stakeholders.
Radio-echo sounding at Dome C, East Antarctica: A comparison of measured and modeled data
NASA Astrophysics Data System (ADS)
Winter, Anna; Eisen, Olaf; Steinhage, Daniel; Zirizzotti, Achille; Urbini, Stefano; Cavitte, Marie; Blankenship, Donald D.; Wolff, Eric
2016-04-01
The internal layering architecture of ice sheets, detected with radio-echo sounding (RES), contains clues to past ice-flow dynamics and mass balance. A common way of relating the recorded travel time of RES reflections to depth is by integrating a wave-speed distribution. This results in an increasing absolute error with depth. We present a synchronization of RES-internal layers of different radar systems (Alfred Wegener Institute, Center for Remote Sensing of Ice Sheets, Istituto Nazionale di Geofisica e Vulcanologia, British Antarctic Survey and University of Texas Institute for Geophysics) with ice-core records from the Antarctic deep drill site Dome C. Synthetic radar traces are obtained from measurements of ice-core density and conductivity with a 1D model of Maxwell's equations. The reflection peaks of the different radar systems' measurements are shifted by a wiggle-matching algorithm, so they match the synthetic trace. In this way, we matched pronounced internal reflections in the RES data to conductivity peaks with considerably smaller depth uncertainties, and assigned them with the ice-core age. We examine the differences in shifts and resolution of the different RES data to address the question of their comparability and combined analysis for an extensive age-depth distribution.
Collection of Light From an Optical Fiber With a Numerical Aperture Greater Than One
NASA Technical Reports Server (NTRS)
Egalon, Claudio O. (Inventor); Rogowski, Robert S. (Inventor)
1996-01-01
In an optical fiber having NA greater than 1, light may be internally reflected when it strikes the fiber end at a fiber-air interface. This problem may be overcome by modification of the fiber by reverse tapering the core. Light is redirected by the taper to strike the interface at an angle closer to normal. This allows light to exit the fiber end that would by internally reflected in an untapered fiber of NA greater than 1. The novelty of the present invention lies in the tapering of the fiber core for increased through transmission of light. Prior art devices have made use of fiber tapers to achieve mode control or fiber coupling. The problem of internal reflection has not been addressed as it is one that is not as important in fibers having NA less than 1, which are more common. In chemical sensing it is advantageous to make use of fibers having higher NA due to an increased sensitivity. However the advantages in sensitivity are diminished due to the loss of signal at the fiber-air interface. The present invention overcomes the problem of loss at the interface, thus facilitating the use of high NA fibers for chemical sensing.
NASA Astrophysics Data System (ADS)
Fujinami, Taku; Kigami, Hiroshi; Unno, Noriyuki; Taniguchi, Jun; Satake, Shin-ichi
2018-03-01
Total internal reflection fluorescence microscopy (TIRFM) is a promising method for measuring fluid flow close to a wall with nanoscale resolution in a process that is termed "multilayer nanoparticle image velocimetry" (MnPIV). TIRFM uses evanescent light that is generated on a substrate (typically a glass slide) by total internal reflection of light. Many researchers have previously studied x-y-z (3D) flows of water close to flat glass slides using MnPIV. On the other hand, a fluid flow close to a structured surface is also important. To measure flows of water near micro-patterns, we previously developed an MnPIV technique that uses a refractive-index-matching method. In previous study, the micropattern is made of a thermoplastic material with a refractive index that closely matches that of water. In this study, ultraviolet nanoimprint lithography was used for fabricating the appropriate micro-patterns because this technique can fabricate a pattern with a high resolution. As a result, we succeeded in performing MnPIV in water with a circular hole array pattern made by ultraviolet nanoimprint using a refractive-index-matching method. We believe that this technique will be helpful in elucidating fluid flows around microstructures.
NASA Astrophysics Data System (ADS)
Korenev, V. V.; Savelyev, A. V.; Zhukov, A. E.; Maximov, M. V.
2015-11-01
The ways to optimize key parameters of active region and edge reflectivity of edge- emitting semiconductor quantum dot laser are provided. It is shown that in the case of optimal cavity length and sufficiently large dispersion lasing spectrum of a given width can be obtained at injection current up to an order of magnitude lower in comparison to non-optimized sample. The influence of internal loss and edge reflection is also studied in details.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Steiner, Myles A.; Perl, E. E.; Geisz, J. F.
Here, we demonstrate that in solar cells with highly reflective back mirrors, the measured internal quantum efficiency exhibits a shift in bandgap relative to the measured external quantum efficiency. The shift arises from the fact that the measured reflectance at the front surface includes a superposition of waves reflecting from the front and back surfaces. We quantify the magnitude of the apparent shift and discuss the errors that can result in determination of quantities such as the photocurrent. Because of this apparent shift, it is important that the bandgap be determined from the external quantum efficiency.
The IAB Congress 2016: Is there justice in the bioethics world?
Gopichandran, Vijayaprasad
2017-01-01
This comment contains the reflections of a first-time participant in the 13th International Association of Bioethics Congress (IAB 2016), held from June 14 to 17, 2016, in Edinburgh. At the outset, I would like to make a couple of clarifications. First, the opinions expressed here are my personal reflections and second, I am a physician and public health practitioner by profession and my interest is bioethics. I reflect on the justice implications of the IAB 2016 from the perspective of the challenge of maintaining inclusivity in a multidisciplinary bioethics world.
[Sources of leader's confidence in organizations].
Ikeda, Hiroshi; Furukawa, Hisataka
2006-04-01
The purpose of this study was to examine the sources of confidence that organization leaders had. As potential sources of the confidence, we focused on fulfillment of expectations made by self and others, reflection on good as well as bad job experiences, and awareness of job experiences in terms of commonality, differentiation, and multiple viewpoints. A questionnaire was administered to 170 managers of Japanese companies. Results were as follows: First, confidence in leaders was more strongly related to fulfillment of expectations made by self and others than reflection on and awareness of job experiences. Second, the confidence was weakly related to internal processing of job experiences, in the form of commonality awareness and reflection on good job experiences. And finally, years of managerial experiences had almost no relation to the confidence. These findings suggested that confidence in leaders was directly acquired from fulfillment of expectations made by self and others, rather than indirectly through internal processing of job experiences. Implications of the findings for leadership training were also discussed.
Development of a high efficiency thin silicon solar cell. [fabrication and stability tests
NASA Technical Reports Server (NTRS)
Lindmayer, J.
1976-01-01
One hundred thin (120 microns to 260 microns) silicon-aluminum solar cells were fabricated and tested. Silicon slices were prepared, into which an aluminum alloy was evaporated over a range of temperatures and times. Antireflection coatings of tantalum oxide were applied to the cells. Reflectance of the silicon-aluminum interfaces was correlated to alloy temperature (graphs are shown). Optical measurements of the rear surface-internal reflectance of the cells were performed using a Beckman spectrophotometer. An improved gridline pattern was evaluated and stability tests (thermal cycling tests) were performed. Results show that: (1) a high-index, high-transmittance antireflection coating was obtained; (2) the improved metallization of the cells gave a 60 percent rear surface-internal reflectance, and the cells displayed excellent fill factors and blue response of the spectrum; (3) an improved gridline pattern (5 micron linewidths compared to 13 micron linewidths) resulted in a 1.3 percent improvement in short circuit currents; and (4) the stability tests showed no change in cell properties.
Pârvan, Alexandra
2016-01-01
Attachment research shows that the formation of unconscious, insecure representations of the self, the other, and the self-other relations is linked to perpetration and receipt of violence. Attachment-focused therapy aims to change these internal schemata to more secure, adaptive representations by therapeutic work addressed to senses, emotions, and behavior. The paper proposes a new approach to altering the self and other representations in offenders and victims: it involves intellectual reflection on self, will, action and responsibility informed by Augustine’s views, facilitated by actual relational experience, and translated into a distinct self-soothing strategy. The reflective-experiential approach can complement existing methods of working with violent or traumatized individuals both within and outside an attachment theory framework. It consists in: identifying that a non-reflective nondistinction between self and behavior supports damaging self- and other- representations and interactions; proposing ways for clients to comprehend and consciously operate with the distinction between self and action. PMID:28936108
NASA Astrophysics Data System (ADS)
Liu, Jiafu; McInnes, Colin R.
2018-03-01
This paper considers utilizing solar radiation pressure (SRP) to actively control the surface shape of a reflector consisting of a rigid hoop and slack membrane with embedded reflectivity control devices. The full nonlinear static partial differential governing equations for a reflector with negligible elastic deformations are established for the circumferential, radial and transverse directions respectively, in which the SRP force with ideal/non-perfect models, the centripetal force caused by the rotation of the reflector and the internal stresses are considered. The inverse problem is then formulated by assuming that the required surface shape is known, and then the governing algebraic-differential equations used to determine the required surface reflectivity, together with the internal stresses where are presented accordingly. The validity of the approach is verified by comparing the results in this paper with corresponding published results as benchmarks. The feasible regions of the angular velocity and Sun angle for a paraboloidal reflector with an invariant radius and focal length (case 1), and the achievable focal lengths with a specific angular velocity and Sun angle (case 2) are presented for two SRP models respectively, both by considering the constraints on the reflectivity and internal stresses. It is then found that the feasible region is toward a larger angular velocity and Sun angle when using the non-perfect SRP model, compared with the ideal one in case 1. The angular velocity of the spinning reflector should be within a certain range to make the required reflectivity profiles within a practical range, i.e., [0, 0.88], as indicated from prior NASA solar sail studies. In case 2, it is found that the smallest achievable focal length of the reflector with the non-perfect SRP model is smaller than that with the ideal SRP model. It is also found that the stress level is extremely low for all cases considered and that the typical real material strength available for the reflector is sufficient to withstand these internal stresses.
Development of internal models and predictive abilities for visual tracking during childhood
Ego, Caroline; Yüksel, Demet
2015-01-01
The prediction of the consequences of our own actions through internal models is an essential component of motor control. Previous studies showed improvement of anticipatory behaviors with age for grasping, drawing, and postural control. Since these actions require visual and proprioceptive feedback, these improvements might reflect both the development of internal models and the feedback control. In contrast, visual tracking of a temporarily invisible target gives specific markers of prediction and internal models for eye movements. Therefore, we recorded eye movements in 50 children (aged 5–19 yr) and in 10 adults, who were asked to pursue a visual target that is temporarily blanked. Results show that the youngest children (5–7 yr) have a general oculomotor behavior in this task, qualitatively similar to the one observed in adults. However, the overall performance of older subjects in terms of accuracy at target reappearance and variability in their behavior was much better than the youngest children. This late maturation of predictive mechanisms with age was reflected into the development of the accuracy of the internal models governing the synergy between the saccadic and pursuit systems with age. Altogether, we hypothesize that the maturation of the interaction between smooth pursuit and saccades that relies on internal models of the eye and target displacement is related to the continuous maturation of the cerebellum. PMID:26510757
Development of internal models and predictive abilities for visual tracking during childhood.
Ego, Caroline; Yüksel, Demet; Orban de Xivry, Jean-Jacques; Lefèvre, Philippe
2016-01-01
The prediction of the consequences of our own actions through internal models is an essential component of motor control. Previous studies showed improvement of anticipatory behaviors with age for grasping, drawing, and postural control. Since these actions require visual and proprioceptive feedback, these improvements might reflect both the development of internal models and the feedback control. In contrast, visual tracking of a temporarily invisible target gives specific markers of prediction and internal models for eye movements. Therefore, we recorded eye movements in 50 children (aged 5-19 yr) and in 10 adults, who were asked to pursue a visual target that is temporarily blanked. Results show that the youngest children (5-7 yr) have a general oculomotor behavior in this task, qualitatively similar to the one observed in adults. However, the overall performance of older subjects in terms of accuracy at target reappearance and variability in their behavior was much better than the youngest children. This late maturation of predictive mechanisms with age was reflected into the development of the accuracy of the internal models governing the synergy between the saccadic and pursuit systems with age. Altogether, we hypothesize that the maturation of the interaction between smooth pursuit and saccades that relies on internal models of the eye and target displacement is related to the continuous maturation of the cerebellum. Copyright © 2016 the American Physiological Society.
Nie, Aiqing; Jiang, Jingguo; Fu, Qiao
2014-08-20
Previous research has found that conjunction faces (whose internal features, e.g. eyes, nose, and mouth, and external features, e.g. hairstyle and ears, are from separate studied faces) and feature faces (partial features of these are studied) can produce higher false alarms than both old and new faces (i.e. those that are exactly the same as the studied faces and those that have not been previously presented) in recognition. The event-related potentials (ERPs) that relate to conjunction and feature faces at recognition, however, have not been described as yet; in addition, the contributions of different facial features toward ERPs have not been differentiated. To address these issues, the present study compared the ERPs elicited by old faces, conjunction faces (the internal and the external features were from two studied faces), old internal feature faces (whose internal features were studied), and old external feature faces (whose external features were studied) with those of new faces separately. The results showed that old faces not only elicited an early familiarity-related FN400, but a more anterior distributed late old/new effect that reflected recollection. Conjunction faces evoked similar late brain waveforms as old internal feature faces, but not to old external feature faces. These results suggest that, at recognition, old faces hold higher familiarity than compound faces in the profiles of ERPs and internal facial features are more crucial than external ones in triggering the brain waveforms that are characterized as reflecting the result of familiarity.
Integrating sphere based reflectance measurements for small-area semiconductor samples
NASA Astrophysics Data System (ADS)
Saylan, S.; Howells, C. T.; Dahlem, M. S.
2018-05-01
This article describes a method that enables reflectance spectroscopy of small semiconductor samples using an integrating sphere, without the use of additional optical elements. We employed an inexpensive sample holder to measure the reflectance of different samples through 2-, 3-, and 4.5-mm-diameter apertures and applied a mathematical formulation to remove the bias from the measured spectra caused by illumination of the holder. Using the proposed method, the reflectance of samples fabricated using expensive or rare materials and/or low-throughput processes can be measured. It can also be incorporated to infer the internal quantum efficiency of small-area, research-level solar cells. Moreover, small samples that reflect light at large angles and develop scattering may also be measured reliably, by virtue of an integrating sphere insensitive to directionalities.
ERIC Educational Resources Information Center
Campbell, Coral J. L.; Walta, Caroline
2015-01-01
Short-term international practicum experience is now a feature of many university education programs in Australia in an attempt to engage students with the growing multi-cultural aspects of Australian life. The stated purposes of such practicum experiences generally highlight intercultural learning, which is associated with the development of…
ERIC Educational Resources Information Center
Patterson, Timothy
2013-01-01
This study is an examination of the stories four social studies teachers told after participating in one international professional development program. Drawing on theories of postcolonialism, this narrative inquiry uses interviews, observations, and artifacts to investigate if and to what degree travel to and study in China influences the…
ERIC Educational Resources Information Center
Peters, Michael A.
2006-01-01
This article charts the rise of global science and a global science infrastructure as part of the emerging international knowledge system exemplifying a geography of knowledge and the importance of new info-communications networks. The article theorises the rise of global science, which still strongly reflects a Western bias and is highly…
ERIC Educational Resources Information Center
Batey, Jacqueline J.; Lupi, Marsha H.
2012-01-01
The study-abroad internship option for students is one example of a transformational learning opportunity (TLO) that is becoming increasingly popular in programs offered by colleges and universities in the United States (Alfaro, 2008; Cushner & Mahon, 2002). These TLOs often have the potential to broaden, enrich, or augment student learning and…
ERIC Educational Resources Information Center
Olson, Elizabeth A.
This report provides brief reflections on the major themes of this conference at which approximately 100 academicians, practitioners, students, and administrators were present. In the opening session, Wilbert J. LeMelle stressed the importance of thinking about a developing world when planning for international educational exchange at the adult…
Children's Perspective on Learning: An International Study in Denmark, Estonia, Germany and Sweden
ERIC Educational Resources Information Center
Sandberg, Anette; Broström, Stig; Johansson, Inge; Frøkjaer, Thorleif; Kieferle, Christa; Seifert, Anja; Roth, Angela; Tuul, Maire; Ugaste, Aino; Laan, Meeli
2017-01-01
This article explores how some children in Denmark, Estonia, Germany and Sweden describe their perspective on learning. The aim of the international study is to gain knowledge of how preschool children in Sweden, Denmark, Estonia and Germany reflect and perceive their learning in preschool and other surrounding social contexts. The results are…
ERIC Educational Resources Information Center
Irish, Tomás
2016-01-01
In 1924 the Carnegie Endowment for International Peace published a volume investigating the teaching of school history in former belligerent states in Europe. The project sought to reconcile former enemies through mutual understanding and educational exchange and reflected a widely held belief that although the military conflict had finished, its…