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Sample records for cell tracking velocimetry

  1. 3D scanning particle tracking velocimetry

    Hoyer, Klaus; Holzner, Markus; Guala, Michele; Liberzon, Alexander; Kinzelbach, Wolfgang [Swiss Federal Institut of Technology Zurich, Institut fuer Hydromechanik und Wasserwirtschaft, Zuerich (Switzerland); Luethi, Beat [Risoe National Laboratory, Roskilde (Denmark)

    2005-11-01

    In this article, we present an experimental setup and data processing schemes for 3D scanning particle tracking velocimetry (SPTV), which expands on the classical 3D particle tracking velocimetry (PTV) through changes in the illumination, image acquisition and analysis. 3D PTV is a flexible flow measurement technique based on the processing of stereoscopic images of flow tracer particles. The technique allows obtaining Lagrangian flow information directly from measured 3D trajectories of individual particles. While for a classical PTV the entire region of interest is simultaneously illuminated and recorded, in SPTV the flow field is recorded by sequential tomographic high-speed imaging of the region of interest. The advantage of the presented method is a considerable increase in maximum feasible seeding density. Results are shown for an experiment in homogenous turbulence and compared with PTV. SPTV yielded an average 3,500 tracked particles per time step, which implies a significant enhancement of the spatial resolution for Lagrangian flow measurements. (orig.)

  2. Streamflow Observations From Cameras: Large-Scale Particle Image Velocimetry or Particle Tracking Velocimetry?

    Tauro, F.; Piscopia, R.; Grimaldi, S.

    2017-12-01

    Image-based methodologies, such as large scale particle image velocimetry (LSPIV) and particle tracking velocimetry (PTV), have increased our ability to noninvasively conduct streamflow measurements by affording spatially distributed observations at high temporal resolution. However, progress in optical methodologies has not been paralleled by the implementation of image-based approaches in environmental monitoring practice. We attribute this fact to the sensitivity of LSPIV, by far the most frequently adopted algorithm, to visibility conditions and to the occurrence of visible surface features. In this work, we test both LSPIV and PTV on a data set of 12 videos captured in a natural stream wherein artificial floaters are homogeneously and continuously deployed. Further, we apply both algorithms to a video of a high flow event on the Tiber River, Rome, Italy. In our application, we propose a modified PTV approach that only takes into account realistic trajectories. Based on our findings, LSPIV largely underestimates surface velocities with respect to PTV in both favorable (12 videos in a natural stream) and adverse (high flow event in the Tiber River) conditions. On the other hand, PTV is in closer agreement than LSPIV with benchmark velocities in both experimental settings. In addition, the accuracy of PTV estimations can be directly related to the transit of physical objects in the field of view, thus providing tangible data for uncertainty evaluation.

  3. Microparticle tracking velocimetry as a tool for microfluidic flow measurements

    Salipante, Paul; Hudson, Steven D.; Schmidt, James W.; Wright, John D.

    2017-07-01

    The accurate measurement of flows in microfluidic channels is important for commercial and research applications. We compare the accuracy of flow measurement techniques over a wide range flows. Flow measurements made using holographic microparticle tracking velocimetry (µPTV) and a gravimetric flow standard over the range of 0.5-100 nL/s agree within 0.25%, well within the uncertainty of the two flow systems. Two commercial thermal flow sensors were used as the intermediaries (transfer standards) between the two flow measurement systems. The gravimetric flow standard was used to calibrate the thermal flow sensors by measuring the rate of change of the mass of liquid in a beaker on a micro-balance as it fills. The holographic µPTV flow measurements were made in a rectangular channel and the flow was seeded with 1 µm diameter polystyrene spheres. The volumetric flow was calculated using the Hagen-Pouiseille solution for a rectangular channel. The uncertainty of both flow measurement systems is given. For the gravimetric standard, relative uncertainty increased for decreasing flows due to surface tension forces between the pipette carrying the flow and the free surface of the liquid in the beaker. The uncertainty of the holographic µPTV measurements did not vary significantly over the measured flow range, and thus comparatively are especially useful at low flow velocities.

  4. Comparative assessment of pressure field reconstructions from particle image velocimetry measurements and Lagrangian particle tracking

    van Gent, P.L.; Michaelis, D; van Oudheusden, B.W.; Weiss, P.E.; de Kat, R.; Laskari, A.; Jeon, Y.J.; David, L; Schanz, D; Huhn, F.; Gesemann, S; Novara, M.; McPhaden, C.; Neeteson, N. J.; Rival, David E.; Schneiders, J.F.G.; Schrijer, F.F.J.

    2017-01-01

    A test case for pressure field reconstruction from particle image velocimetry (PIV) and Lagrangian particle tracking (LPT) has been developed by constructing a simulated experiment from a zonal detached eddy simulation for an axisymmetric base flow at Mach 0.7. The test case comprises sequences

  5. Track benchmarking method for uncertainty quantification of particle tracking velocimetry interpolations

    Schneiders, Jan F G; Sciacchitano, Andrea

    2017-01-01

    The track benchmarking method (TBM) is proposed for uncertainty quantification of particle tracking velocimetry (PTV) data mapped onto a regular grid. The method provides statistical uncertainty for a velocity time-series and can in addition be used to obtain instantaneous uncertainty at increased computational cost. Interpolation techniques are typically used to map velocity data from scattered PTV (e.g. tomographic PTV and Shake-the-Box) measurements onto a Cartesian grid. Recent examples of these techniques are the FlowFit and VIC+  methods. The TBM approach estimates the random uncertainty in dense velocity fields by performing the velocity interpolation using a subset of typically 95% of the particle tracks and by considering the remaining tracks as an independent benchmarking reference. In addition, also a bias introduced by the interpolation technique is identified. The numerical assessment shows that the approach is accurate when particle trajectories are measured over an extended number of snapshots, typically on the order of 10. When only short particle tracks are available, the TBM estimate overestimates the measurement error. A correction to TBM is proposed and assessed to compensate for this overestimation. The experimental assessment considers the case of a jet flow, processed both by tomographic PIV and by VIC+. The uncertainty obtained by TBM provides a quantitative evaluation of the measurement accuracy and precision and highlights the regions of high error by means of bias and random uncertainty maps. In this way, it is possible to quantify the uncertainty reduction achieved by advanced interpolation algorithms with respect to standard correlation-based tomographic PIV. The use of TBM for uncertainty quantification and comparison of different processing techniques is demonstrated. (paper)

  6. Three-dimensional particle tracking velocimetry using dynamic vision sensors

    Borer, D.; Delbruck, T.; Rösgen, T.

    2017-12-01

    A fast-flow visualization method is presented based on tracking neutrally buoyant soap bubbles with a set of neuromorphic cameras. The "dynamic vision sensors" register only the changes in brightness with very low latency, capturing fast processes at a low data rate. The data consist of a stream of asynchronous events, each encoding the corresponding pixel position, the time instant of the event and the sign of the change in logarithmic intensity. The work uses three such synchronized cameras to perform 3D particle tracking in a medium sized wind tunnel. The data analysis relies on Kalman filters to associate the asynchronous events with individual tracers and to reconstruct the three-dimensional path and velocity based on calibrated sensor information.

  7. A new paradigm for particle tracking velocimetry, based on graph-theory and pulsed neural network

    Derou, D.; Herault, L.

    1994-01-01

    The Particle Tracking Velocimetry (PTV) technique works by recording, at different instances in time, positions of small tracers particles following a flow and illuminated by a sheet, or pseudo sheet, of light. It aims to recognize each particle trajectory, constituted of n different spots and determine thus each particle velocity vector. In this paper, we devise a new method, taking into account a global consistency of the trajectories to be extracted, in terms of visual perception and physical properties. It is based on a graph-theoretic formulation of the particle tracking problem and the use of an original neural network, called pulsed neural network. (authors). 4 figs

  8. Development of two-dimensional velocity field measurement using particle tracking velocimetry on neutron radiography

    Saito, Y.; Mishima, K.; Suzuki, T.; Matsubayashi, M.

    2003-01-01

    The structures of liquid metal two-phase flow are investigated for analyzing the core meltdown accident of fast reactor. The experiments of high-density ratio two-phase flow for lead-bismuth molten metal and nitrogen gases are conducted to understand in detail. The liquid phase velocity distributions of lead-bismuth molten metal are measured by neutron radiography using Au-Cd tracer particles. The liquid phase velocity distributions are obtained usually by using particle image velocimetry (PIV) on the neutron radiography. The PIV, however is difficult to get the velocity vector distribution quantitatively. An image of neutron radiography is divided into two images of the bubbles and the tracer particles each in particle tracking velocimetry (PTV), which distinguishes tracer contents in the bubble from them in the liquid phase. The locations of tracer particles in the liquid phase are possible to determine by particle mask correlation method, in which the bubble images are separated from the tracer images by Σ-scaling method. The particle tracking velocimetry give a full detail of the velocity vector distributions of the liquid phase in two-phase flow, in comparison with the PIV method. (M. Suetake)

  9. Improvement in the independence of relaxation method-based particle tracking velocimetry

    Jia, P; Wang, Y; Zhang, Y

    2013-01-01

    New techniques are developed to improve the independence of relaxation method-based particle tracking velocimetry (RM-PTV). Firstly, Delaunay tessellation (DT) is employed to form clusters of neighboring particles with similar motion in the same frame; and then a bidirectional calculation concept is adopted to improve the way of particle pairing. These new techniques are tested with both self-defined particle images and the particle image velocimetry standard synthetic particle images. The results indicate that the DT method performs well and efficiently in determining the particle clusters, and the particle pairing process is well optimized by the bidirectional calculation concept. With these methods, three computation parameters are eliminated, which makes RM-PTV more autonomous in applications. (paper)

  10. Measuring the 3D motion of particles in microchannel acoustophoresis using astigmatism particle tracking velocimetry

    Augustsson, P.; Barnkob, Rune; Bruus, Henrik

    2012-01-01

    We introduce full three-dimensional tracking of particles in an acoustophoresis microchannel using Astigmatism Particle Tracking Velocimetry (APTV) [1]. For the first time the interaction between acoustic streaming and the primary acoustic radiation force in microchannel acoustophoresis are exami...... relative to the influence from the acoustic radiation force. The current study opens the route to optimized acoustophoretic system design and operation to enable manipulation of small biological components such as spores, bacteria and viruses.......We introduce full three-dimensional tracking of particles in an acoustophoresis microchannel using Astigmatism Particle Tracking Velocimetry (APTV) [1]. For the first time the interaction between acoustic streaming and the primary acoustic radiation force in microchannel acoustophoresis...... are examined in three dimensions. We have quantified the velocity of particles driven by the primary acoustic radiation force and acoustic streaming, respectively, using 0.5-μm and 5-μm particles. Increased ultrasound frequency and lowered viscosity of the medium reduced the influence of acoustic streaming...

  11. Laboratory observations of sediment transport using combined particle image and tracking velocimetry (Conference Presentation)

    Frank, Donya; Calantoni, Joseph

    2017-05-01

    Improved understanding of coastal hydrodynamics and morphology will lead to more effective mitigation measures that reduce fatalities and property damage caused by natural disasters such as hurricanes. We investigated sediment transport under oscillatory flow over flat and rippled beds with phase-separated stereoscopic Particle Image Velocimetry (PIV). Standard PIV techniques severely limit measurements at the fluid-sediment interface and do not allow for the observation of separate phases in multi-phase flow (e.g. sand grains in water). We have implemented phase-separated Particle Image Velocimetry by adding fluorescent tracer particles to the fluid in order to observe fluid flow and sediment transport simultaneously. While sand grains scatter 532 nm wavelength laser light, the fluorescent particles absorb 532 nm laser light and re-emit light at a wavelength of 584 nm. Optical long-pass filters with a cut-on wavelength of 550 nm were installed on two cameras configured to perform stereoscopic PIV to capture only the light emitted by the fluorescent tracer particles. A third high-speed camera was used to capture the light scattered by the sand grains allowing for sediment particle tracking via particle tracking velocimetry (PTV). Together, these overlapping, simultaneously recorded images provided sediment particle and fluid velocities at high temporal and spatial resolution (100 Hz sampling with 0.8 mm vector spacing for the 2D-3C fluid velocity field). Measurements were made under a wide range of oscillatory flows over flat and rippled sand beds. The set of observations allow for the investigation of the relative importance of pressure gradients and shear stresses on sediment transport.

  12. Particle Data Management Software for 3DParticle Tracking Velocimetry and Related Applications – The Flowtracks Package

    Yosef Meller

    2016-06-01

    Full Text Available The Particle Tracking Velocimetry (PTV community employs several formats of particle information such as position and velocity as function of time, i.e. trajectory data, as a result of diverging needs unmet by existing formats, and a number of different, mostly home-grown, codes for handling the data. Flowtracks is a Python package that provides a single code base for accessing different formats as a database, i.e. storing data and programmatically manipulating them using format-agnostic data structures. Furthermore, it offers an HDF5-based format that is fast and extensible, obviating the need for other formats. The package may be obtained from https://github.com/OpenPTV/postptv and used as-is by many fluid-dynamics labs, or with minor extensions adhering to a common interface, by researchers from other fields, such as biology and population tracking.

  13. Droplet morphometry and velocimetry (DMV): a video processing software for time-resolved, label-free tracking of droplet parameters.

    Basu, Amar S

    2013-05-21

    Emerging assays in droplet microfluidics require the measurement of parameters such as drop size, velocity, trajectory, shape deformation, fluorescence intensity, and others. While micro particle image velocimetry (μPIV) and related techniques are suitable for measuring flow using tracer particles, no tool exists for tracking droplets at the granularity of a single entity. This paper presents droplet morphometry and velocimetry (DMV), a digital video processing software for time-resolved droplet analysis. Droplets are identified through a series of image processing steps which operate on transparent, translucent, fluorescent, or opaque droplets. The steps include background image generation, background subtraction, edge detection, small object removal, morphological close and fill, and shape discrimination. A frame correlation step then links droplets spanning multiple frames via a nearest neighbor search with user-defined matching criteria. Each step can be individually tuned for maximum compatibility. For each droplet found, DMV provides a time-history of 20 different parameters, including trajectory, velocity, area, dimensions, shape deformation, orientation, nearest neighbour spacing, and pixel statistics. The data can be reported via scatter plots, histograms, and tables at the granularity of individual droplets or by statistics accrued over the population. We present several case studies from industry and academic labs, including the measurement of 1) size distributions and flow perturbations in a drop generator, 2) size distributions and mixing rates in drop splitting/merging devices, 3) efficiency of single cell encapsulation devices, 4) position tracking in electrowetting operations, 5) chemical concentrations in a serial drop dilutor, 6) drop sorting efficiency of a tensiophoresis device, 7) plug length and orientation of nonspherical plugs in a serpentine channel, and 8) high throughput tracking of >250 drops in a reinjection system. Performance metrics

  14. A multi-time-step noise reduction method for measuring velocity statistics from particle tracking velocimetry

    Machicoane, Nathanaël; López-Caballero, Miguel; Bourgoin, Mickael; Aliseda, Alberto; Volk, Romain

    2017-10-01

    We present a method to improve the accuracy of velocity measurements for fluid flow or particles immersed in it, based on a multi-time-step approach that allows for cancellation of noise in the velocity measurements. Improved velocity statistics, a critical element in turbulent flow measurements, can be computed from the combination of the velocity moments computed using standard particle tracking velocimetry (PTV) or particle image velocimetry (PIV) techniques for data sets that have been collected over different values of time intervals between images. This method produces Eulerian velocity fields and Lagrangian velocity statistics with much lower noise levels compared to standard PIV or PTV measurements, without the need of filtering and/or windowing. Particle displacement between two frames is computed for multiple different time-step values between frames in a canonical experiment of homogeneous isotropic turbulence. The second order velocity structure function of the flow is computed with the new method and compared to results from traditional measurement techniques in the literature. Increased accuracy is also demonstrated by comparing the dissipation rate of turbulent kinetic energy measured from this function against previously validated measurements.

  15. Particle Image Velocimetry and Computational Fluid Dynamics Analysis of Fuel Cell Manifold

    Lebæk, Jesper; Blazniak Andreasen, Marcin; Andresen, Henrik Assenholm

    2010-01-01

    The inlet effect on the manifold flow in a fuel cell stack was investigated by means of numerical methods (computational fluid dynamics) and experimental methods (particle image velocimetry). At a simulated high current density situation the flow field was mapped on a 70 cell simulated cathode...

  16. Radial basis function interpolation of unstructured, three-dimensional, volumetric particle tracking velocimetry data

    Casa, L D C; Krueger, P S

    2013-01-01

    Unstructured three-dimensional fluid velocity data were interpolated using Gaussian radial basis function (RBF) interpolation. Data were generated to imitate the spatial resolution and experimental uncertainty of a typical implementation of defocusing digital particle image velocimetry. The velocity field associated with a steadily rotating infinite plate was simulated to provide a bounded, fully three-dimensional analytical solution of the Navier–Stokes equations, allowing for robust analysis of the interpolation accuracy. The spatial resolution of the data (i.e. particle density) and the number of RBFs were varied in order to assess the requirements for accurate interpolation. Interpolation constraints, including boundary conditions and continuity, were included in the error metric used for the least-squares minimization that determines the interpolation parameters to explore methods for improving RBF interpolation results. Even spacing and logarithmic spacing of RBF locations were also investigated. Interpolation accuracy was assessed using the velocity field, divergence of the velocity field, and viscous torque on the rotating boundary. The results suggest that for the present implementation, RBF spacing of 0.28 times the boundary layer thickness is sufficient for accurate interpolation, though theoretical error analysis suggests that improved RBF positioning may yield more accurate results. All RBF interpolation results were compared to standard Gaussian weighting and Taylor expansion interpolation methods. Results showed that RBF interpolation improves interpolation results compared to the Taylor expansion method by 60% to 90% based on the average squared velocity error and provides comparable velocity results to Gaussian weighted interpolation in terms of velocity error. RMS accuracy of the flow field divergence was one to two orders of magnitude better for the RBF interpolation compared to the other two methods. RBF interpolation that was applied to

  17. 3D particle tracking velocimetry using dynamic discrete tomography for plasma physics applications

    Moseev, Dmitry; Alpers, Andreas; Gritzmann, Peter

    2013-01-01

    tomography algorithm is efficient for data from two projection directions and exact. The non-uniqueness can be detected and tracked individually. The algorithm performance is proportional to N3 on average where N is the number of particles in the reconstruction. There is a room for further improvement...

  18. Parallel computing of a digital hologram and particle searching for microdigital-holographic particle-tracking velocimetry

    Satake, Shin-ichi; Kanamori, Hiroyuki; Kunugi, Tomoaki; Sato, Kazuho; Ito, Tomoyoshi; Yamamoto, Keisuke

    2007-01-01

    We have developed a parallel algorithm for microdigital-holographic particle-tracking velocimetry. The algorithm is used in (1) numerical reconstruction of a particle image computer using a digital hologram, and (2) searching for particles. The numerical reconstruction from the digital hologram makes use of the Fresnel diffraction equation and the FFT (fast Fourier transform),whereas the particle search algorithm looks for local maximum graduation in a reconstruction field represented by a 3D matrix. To achieve high performance computing for both calculations (reconstruction and particle search), two memory partitions are allocated to the 3D matrix. In this matrix, the reconstruction part consists of horizontally placed 2D memory partitions on the x-y plane for the FFT, whereas, the particle search part consists of vertically placed 2D memory partitions set along the z axes.Consequently, the scalability can be obtained for the proportion of processor elements,where the benchmarks are carried out for parallel computation by a SGI Altix machine

  19. Rectification of Image Velocity Results (RIVeR): A simple and user-friendly toolbox for large scale water surface Particle Image Velocimetry (PIV) and Particle Tracking Velocimetry (PTV)

    Patalano, Antoine; García, Carlos Marcelo; Rodríguez, Andrés

    2017-12-01

    LSPIV (Large Scale Particle Image Velocimetry) and LSPTV (Large Scale Particle Tracking Velocimetry) are used as relatively low-cost and non-intrusive techniques for water-surface velocity analysis and flow discharge measurements in rivers or large-scale hydraulic models. This paper describes a methodology based on state-of-the-art tools (for example, that apply classical PIV/PTV analysis) resulting in large-scale surface-flow characterization according to the first operational version of the RIVeR (Rectification of Image Velocity Results). RIVeR is developed in Matlab and is designed to be user-friendly. RIVeR processes large-scale water-surface characterization such as velocity fields or individual trajectories of floating tracers. This work describes the wide range of application of the techniques for comparing measured surface flows in hydraulic physical models to flow discharge estimates for a wide range of flow events in rivers (for example, low and high flows).

  20. Capillary red blood cell velocimetry by phase-resolved optical coherence tomography.

    Tang, Jianbo; Erdener, Sefik Evren; Fu, Buyin; Boas, David A

    2017-10-01

    We present a phase-resolved optical coherence tomography (OCT) method to extend Doppler OCT for the accurate measurement of the red blood cell (RBC) velocity in cerebral capillaries. OCT data were acquired with an M-mode scanning strategy (repeated A-scans) to account for the single-file passage of RBCs in a capillary, which were then high-pass filtered to remove the stationary component of the signal to ensure an accurate measurement of phase shift of flowing RBCs. The angular frequency of the signal from flowing RBCs was then quantified from the dynamic component of the signal and used to calculate the axial speed of flowing RBCs in capillaries. We validated our measurement by RBC passage velocimetry using the signal magnitude of the same OCT time series data.

  1. Particle image velocimetry (PIV) study of rotating cylindrical filters for animal cell perfusion processes.

    Figueredo-Cardero, Alvio; Chico, Ernesto; Castilho, Leda; de Andrade Medronho, Ricardo

    2012-01-01

    In the present work, the main fluid flow features inside a rotating cylindrical filtration (RCF) system used as external cell retention device for animal cell perfusion processes were investigated using particle image velocimetry (PIV). The motivation behind this work was to provide experimental fluid dynamic data for such turbulent flow using a high-permeability filter, given the lack of information about this system in the literature. The results shown herein gave evidence that, at the boundary between the filter mesh and the fluid, a slip velocity condition in the tangential direction does exist, which had not been reported in the literature so far. In the RCF system tested, this accounted for a fluid velocity 10% lower than that of the filter tip, which could be important for the cake formation kinetics during filtration. Evidence confirming the existence of Taylor vortices under conditions of turbulent flow and high permeability, typical of animal cell perfusion RCF systems, was obtained. Second-order turbulence statistics were successfully calculated. The radial behavior of the second-order turbulent moments revealed that turbulence in this system is highly anisotropic, which is relevant for performing numerical simulations of this system. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

  2. Cell tracking. Principles and applications

    Grimm, Jan; Kircher, Moritz F.; Weissleder, Ralph

    2007-01-01

    Cell based therapies such as stem cell therapies or adoptive immunotherapies are currently being explored as a potential treatment for a variety of diseases such as Parkinson's disease, diabetes or cancer. However, quantitative and qualitative evaluation of adoptively transferred cells is indispensable for monitoring the efficiency of the treatment. Current approaches mostly analyze transferred cells from peripheral blood, which cannot assess whether transferred cells actuallyhome to and stay in the targeted tissue. Using cell-labeling methods such as direct labeling or transfection with a marker gene in conjunction with various imaging modalities (MRI, optical or nuclear imaging), labeled cells can be followed in vivo in real-time, and their accumulation as well as function in vivo can be monitored and quantified accurately. This method is usually referred to as ''cell tracking'' or ''cell trafficking'' and is also being applied in basic biological sciences, exemplified in the evaluation of genes contributing to metastasis. This review focuses on principles of this promising methodology and explains various approaches by highlighting recent examples. (orig.) [de

  3. Development of a Vision-Based Particle Tracking Velocimetry Method and Post-Processing of Scattered Velocity Data

    2012-01-01

    the best performance. The values in these cells are in a bold underlined font . Table 7.RMS error of strain rate from various interpolation schemes on...Computer Vision & Pattern Recognition Conference, 1997, (pp. 261-266). San Juan , Puerto Rico. 62 46. Ponchaut N, Mouton C. (2005). 3D

  4. Further development of microparticle image velocimetry analysis for characterisation of gas streams as a novel method of fuel cell development. Final report; Weiterentwicklung des Mikro-Particle Image Velocimetry Analyseverfahrens zur Charakterisierung von Gasstroemungen als neuartige Entwicklungsmethodik fuer Brennstoffzellen. Schlussbericht

    NONE

    2012-07-01

    The project aimed at a better understanding of the complex fluid-mechanical processes in the small ducts of bipolar plates. So far, an appropriate technology for in-situ measurement was lacking. The project therefore focused on the further development of microparticle image velocimetry in order to enable analyses of the local velocity distribution of a gas stream in a microduct. Further, measurements were carried out in the microducts of a fuel cell in the more difficult conditions of actual operation. (orig./AKB) [German] Anlass des Forschungsvorhabens war die komplizierten stroemungsmechanischen Zusammenhaenge in den kleinen Kanaelen der Bipolarplatten zu verstehen. Bisher stand keine Messtechnik zur Verfuegung, dies es erlaubt, die stroemungsmechanischen Prozesse in den Mikrokanaelen unter Realbedingungen in situ zu vermessen und mit der instantanen Zellleistung zu korrelieren, Ziel des Projektes war es daher, die Methode der Mikro-Partikel-Image-Velocimetry in der Art weiterzuentwickeln, dass eine Analyse der lokalen Geschwindigkeitsverteilung einer Gasstroemung in einem Mikrokanal ermoeglicht wird. Darueber hinaus wird als zweites Ziel des Projekts eine solche Messung unter den erschwerten Bedingungen einer betriebenen Brennstoffzelle in Mikrokanaelen einer Zelle durchgefuehrt.

  5. Micro-PIV (micro particle image velocimetry) visualization of red blood cells (RBCs) sucked by a female mosquito

    Kikuchi, K; Mochizuki, O

    2011-01-01

    A mosquito's pump is a highly effective system in the small suction domain. To understand a mosquito's blood suction mechanism, we analysed the characteristics of red blood cells (RBCs) in human blood during and after suction by a female mosquito. Focussing on the flow patterns of the RBCs in human blood being sucked by a mosquito, we visualized blood flow by using a micro-particle image velocimetry (μ-PIV) system, which combines an optical microscope and a PIV method. In an ex vivo experiment, a female mosquito was supplied diluted blood at the tip of the proboscis. We examined the blood flow around the tip of the proboscis and observed that RBCs were periodically sucked towards a hole around the tip. The sucked RBCs then homogeneously flowed parallel to the inner surface of the proboscis without adhering to the wall. Furthermore, using a bioelectric recording system, we directly measured electrical signals generated during suction by the pump muscles located in the mosquito's head. We found that the electrical signal power was synchronized with the acceleration of the RBCs in the sucking phase. A histological stain method was adapted for the observation of the form and internal structure of RBCs in the mosquito. Although the blood flow analysis revealed that the RBCs underwent shear stress during suction, RBCs in the mosquito's stomach maintained their original shape

  6. Rainbow Particle Imaging Velocimetry

    Xiong, Jinhui

    2017-04-27

    Despite significant recent progress, dense, time-resolved imaging of complex, non-stationary 3D flow velocities remains an elusive goal. This work tackles this problem by extending an established 2D method, Particle Imaging Velocimetry, to three dimensions by encoding depth into color. The encoding is achieved by illuminating the flow volume with a continuum of light planes (a “rainbow”), such that each depth corresponds to a specific wavelength of light. A diffractive component in the camera optics ensures that all planes are in focus simultaneously. With this setup, a single color camera is sufficient to track 3D trajectories of particles by combining 2D spatial and 1D color information. For reconstruction, this thesis derives an image formation model for recovering stationary 3D particle positions. 3D velocity estimation is achieved with a variant of 3D optical flow that accounts for both physical constraints as well as the rainbow image formation model. The proposed method is evaluated by both simulations and an experimental prototype setup.

  7. In vivo cell tracking with bioluminescence imaging

    Kim, Jung Eun; Kalimuthu, Senthilkumar; Ahn, Byeong Cheol [Dept. of Nuclear Medicine, Kyungpook National University School of Medicine and Hospital, Daegu (Korea, Republic of)

    2015-03-15

    Molecular imaging is a fast growing biomedical research that allows the visual representation, characterization and quantification of biological processes at the cellular and subcellular levels within intact living organisms. In vivo tracking of cells is an indispensable technology for development and optimization of cell therapy for replacement or renewal of damaged or diseased tissue using transplanted cells, often autologous cells. With outstanding advantages of bioluminescence imaging, the imaging approach is most commonly applied for in vivo monitoring of transplanted stem cells or immune cells in order to assess viability of administered cells with therapeutic efficacy in preclinical small animal models. In this review, a general overview of bioluminescence is provided and recent updates of in vivo cell tracking using the bioluminescence signal are discussed.

  8. Pipeline for Tracking Neural Progenitor Cells

    Vestergaard, Jacob Schack; Dahl, Anders Lindbjerg; Holm, Peter

    2012-01-01

    Automated methods for neural stem cell lineage construction become increasingly important due to the large amount of data produced from time lapse imagery of in vitro cell growth experiments. Segmentation algorithms with the ability to adapt to the problem at hand and robust tracking methods play...... a key role in constructing these lineages. We present here a tracking pipeline based on learning a dictionary of discriminative image patches for segmentation and a graph formulation of the cell matching problem incorporating topology changes and acknowledging the fact that segmentation errors do occur...

  9. Software for precise tracking of cell proliferation

    Kurokawa, Hiroshi; Noda, Hisayori; Sugiyama, Mayu; Sakaue-Sawano, Asako; Fukami, Kiyoko; Miyawaki, Atsushi

    2012-01-01

    Highlights: ► We developed software for analyzing cultured cells that divide as well as migrate. ► The active contour model (Snakes) was used as the core algorithm. ► The time backward analysis was also used for efficient detection of cell division. ► With user-interactive correction functions, the software enables precise tracking. ► The software was successfully applied to cells with fluorescently-labeled nuclei. -- Abstract: We have developed a multi-target cell tracking program TADOR, which we applied to a series of fluorescence images. TADOR is based on an active contour model that is modified in order to be free of the problem of locally optimal solutions, and thus is resistant to signal fluctuation and morphological changes. Due to adoption of backward tracing and addition of user-interactive correction functions, TADOR is used in an off-line and semi-automated mode, but enables precise tracking of cell division. By applying TADOR to the analysis of cultured cells whose nuclei had been fluorescently labeled, we tracked cell division and cell-cycle progression on coverslips over an extended period of time.

  10. Stem cell tracking using iron oxide nanoparticles

    Bull E

    2014-03-01

    Full Text Available Elizabeth Bull,1 Seyed Yazdan Madani,1 Roosey Sheth,1 Amelia Seifalian,1 Mark Green,2 Alexander M Seifalian1,31UCL Centre for Nanotechnology and Regenerative Medicine, Division of Surgery and Interventional Science, University College London, London, 2Department of Physics, King’s College London, Strand Campus, London, UK; 3Royal Free London National Health Service Foundation Trust Hospital, London, UKAbstract: Superparamagnetic iron oxide nanoparticles (SPIONs are an exciting advancement in the field of nanotechnology. They expand the possibilities of noninvasive analysis and have many useful properties, making them potential candidates for numerous novel applications. Notably, they have been shown that they can be tracked by magnetic resonance imaging (MRI and are capable of conjugation with various cell types, including stem cells. In-depth research has been undertaken to establish these benefits, so that a deeper level of understanding of stem cell migratory pathways and differentiation, tumor migration, and improved drug delivery can be achieved. Stem cells have the ability to treat and cure many debilitating diseases with limited side effects, but a main problem that arises is in the noninvasive tracking and analysis of these stem cells. Recently, researchers have acknowledged the use of SPIONs for this purpose and have set out to establish suitable protocols for coating and attachment, so as to bring MRI tracking of SPION-labeled stem cells into common practice. This review paper explains the manner in which SPIONs are produced, conjugated, and tracked using MRI, as well as a discussion on their limitations. A concise summary of recently researched magnetic particle coatings is provided, and the effects of SPIONs on stem cells are evaluated, while animal and human studies investigating the role of SPIONs in stem cell tracking will be explored.Keywords: stem cells, nanoparticle, magnetic

  11. Radiopharmaceutical Stem Cell Tracking for Neurological Diseases

    Paulo Henrique Rosado-de-Castro

    2014-01-01

    Full Text Available Although neurological ailments continue to be some of the main causes of disease burden in the world, current therapies such as pharmacological agents have limited potential in the restoration of neural functions. Cell therapies, firstly applied to treat different hematological diseases, are now being investigated in preclinical and clinical studies for neurological illnesses. However, the potential applications and mechanisms for such treatments are still poorly comprehended and are the focus of permanent research. In this setting, noninvasive in vivo imaging allows better understanding of several aspects of stem cell therapies. Amongst the various methods available, radioisotope cell labeling has become one of the most promising since it permits tracking of cells after injection by different routes to investigate their biodistribution. A significant increase in the number of studies utilizing this method has occurred in the last years. Here, we review the different radiopharmaceuticals, imaging techniques, and findings of the preclinical and clinical reports published up to now. Moreover, we discuss the limitations and future applications of radioisotope cell labeling in the field of cell transplantation for neurological diseases.

  12. Particle Image Velocimetry

    Zhang, Chen; Vasilevskis, Sandijs; Kozlowski, Bartosz

    Particle image velocimetry (PIV) is a non-intrusive, whole filed optical method providing instantaneous velocity information in fluids. The flow is seeded with tracer particles. The particles are illuminated in the target area with a light sheet at least twice within a short time interval....... The camera images the target area and captures each light pulse in separate image frames. The displacement of the particle between the light pulses can be used to determine the velocity vectors. This guideline introduces the principle of the PIV system and the system configuration. The measurement procedure...

  13. Particle-pair relative velocity measurement in high-Reynolds-number homogeneous and isotropic turbulence using 4-frame particle tracking velocimetry

    Dou, Zhongwang; Ireland, Peter J.; Bragg, Andrew D.; Liang, Zach; Collins, Lance R.; Meng, Hui

    2018-02-01

    The radial relative velocity (RV) between particles suspended in turbulent flow plays a critical role in droplet collision and growth. We present a simple and accurate approach to RV measurement in isotropic turbulence—planar 4-frame particle tracking velocimetry—using routine PIV hardware. It improves particle positioning and pairing accuracy over the 2-frame holographic approach by de Jong et al. (Int J Multiphas Flow 36:324-332; de Jong et al., Int J Multiphas Flow 36:324-332, 2010) without using high-speed cameras and lasers as in Saw et al. (Phys Fluids 26:111702, 2014). Homogeneous and isotropic turbulent flow ({R_λ }=357) in a new, fan-driven, truncated iscosahedron chamber was laden with either low-Stokes (mean St=0.09, standard deviation 0.05) or high-Stokes aerosols (mean St=3.46, standard deviation 0.57). For comparison, DNS was conducted under similar conditions ({R_λ }=398; St=0.10 and 3.00, respectively). Experimental RV probability density functions (PDF) and mean inward RV agree well with DNS. Mean inward RV increases with St at small particle separations, r, and decreases with St at large r, indicating the dominance of "path-history" and "inertial filtering" effects, respectively. However, at small r, the experimental mean inward RV trends higher than DNS, possibly due to the slight polydispersity of particles and finite light sheet thickness in experiments. To confirm this interpretation, we performed numerical experiments and found that particle polydispersity increases mean inward RV at small r, while finite laser thickness also overestimates mean inward RV at small r, This study demonstrates the feasibility of accurately measuring RV using routine hardware, and verifies, for the first time, the path-history and inertial filtering effects on particle-pair RV at large particle separations experimentally.

  14. A benchmark for comparison of cell tracking algorithms

    M. Maška (Martin); V. Ulman (Vladimír); K. Svoboda; P. Matula (Pavel); P. Matula (Petr); C. Ederra (Cristina); A. Urbiola (Ainhoa); T. España (Tomás); R. Venkatesan (Rajkumar); D.M.W. Balak (Deepak); P. Karas (Pavel); T. Bolcková (Tereza); M. Štreitová (Markéta); C. Carthel (Craig); S. Coraluppi (Stefano); N. Harder (Nathalie); K. Rohr (Karl); K.E.G. Magnusson (Klas E.); J. Jaldén (Joakim); H.M. Blau (Helen); O.M. Dzyubachyk (Oleh); P. Křížek (Pavel); G.M. Hagen (Guy); D. Pastor-Escuredo (David); D. Jimenez-Carretero (Daniel); M.J. Ledesma-Carbayo (Maria); A. Muñoz-Barrutia (Arrate); E. Meijering (Erik); M. Kozubek (Michal); C. Ortiz-De-Solorzano (Carlos)

    2014-01-01

    textabstractMotivation: Automatic tracking of cells in multidimensional time-lapse fluorescence microscopy is an important task in many biomedical applications. A novel framework for objective evaluation of cell tracking algorithms has been established under the auspices of the IEEE International

  15. 3D Rainbow Particle Tracking Velocimetry

    Aguirre-Pablo, Andres A.; Xiong, Jinhui; Idoughi, Ramzi; Aljedaani, Abdulrahman B.; Dun, Xiong; Fu, Qiang; Thoroddsen, Sigurdur T.; Heidrich, Wolfgang

    2017-11-01

    A single color camera is used to reconstruct a 3D-3C velocity flow field. The camera is used to record the 2D (X,Y) position and colored scattered light intensity (Z) from white polyethylene tracer particles in a flow. The main advantage of using a color camera is the capability of combining different intensity levels for each color channel to obtain more depth levels. The illumination system consists of an LCD projector placed perpendicularly to the camera. Different intensity colored level gradients are projected onto the particles to encode the depth position (Z) information of each particle, benefiting from the possibility of varying the color profiles and projected frequencies up to 60 Hz. Chromatic aberrations and distortions are estimated and corrected using a 3D laser engraved calibration target. The camera-projector system characterization is presented considering size and depth position of the particles. The use of these components reduces dramatically the cost and complexity of traditional 3D-PTV systems.

  16. Track structure model of cell damage in space flight

    Katz, Robert; Cucinotta, Francis A.; Wilson, John W.; Shinn, Judy L.; Ngo, Duc M.

    1992-01-01

    The phenomenological track-structure model of cell damage is discussed. A description of the application of the track-structure model with the NASA Langley transport code for laboratory and space radiation is given. Comparisons to experimental results for cell survival during exposure to monoenergetic, heavy-ion beams are made. The model is also applied to predict cell damage rates and relative biological effectiveness for deep-space exposures.

  17. Automated Tracking of Cell Migration with Rapid Data Analysis.

    DuChez, Brian J

    2017-09-01

    Cell migration is essential for many biological processes including development, wound healing, and metastasis. However, studying cell migration often requires the time-consuming and labor-intensive task of manually tracking cells. To accelerate the task of obtaining coordinate positions of migrating cells, we have developed a graphical user interface (GUI) capable of automating the tracking of fluorescently labeled nuclei. This GUI provides an intuitive user interface that makes automated tracking accessible to researchers with no image-processing experience or familiarity with particle-tracking approaches. Using this GUI, users can interactively determine a minimum of four parameters to identify fluorescently labeled cells and automate acquisition of cell trajectories. Additional features allow for batch processing of numerous time-lapse images, curation of unwanted tracks, and subsequent statistical analysis of tracked cells. Statistical outputs allow users to evaluate migratory phenotypes, including cell speed, distance, displacement, and persistence, as well as measures of directional movement, such as forward migration index (FMI) and angular displacement. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  18. Tracking of stem cells for treatment in cardiovascular disease

    Kang, Won Jun

    2005-01-01

    Various stem cells or progenitor cells are being used to treat cardiovascular disease. In ischemic heart disease, stem cell therapy is expected to regenerate damaged myocardium. To evaluate effects of stem cell treatment, the method to image stem cell location, distribution and differentiation is necessary. Optical imaging, MRI, nuclear imaging methods have been used for tracking stem cells. The methods and problems of each imaging technique are reviewed

  19. Iron Oxide as an MRI Contrast Agent for Cell Tracking

    Korchinski, Daniel J.; Taha, May; Yang, Runze; Nathoo, Nabeela; Dunn, Jeff F.

    2015-01-01

    Iron oxide contrast agents have been combined with magnetic resonance imaging for cell tracking. In this review, we discuss coating properties and provide an overview of ex vivo and in vivo labeling of different cell types, including stem cells, red blood cells, and monocytes/macrophages. Furthermore, we provide examples of applications of cell tracking with iron contrast agents in stroke, multiple sclerosis, cancer, arteriovenous malformations, and aortic and cerebral aneurysms. Attempts at quantifying iron oxide concentrations and other vascular properties are examined. We advise on designing studies using iron contrast agents including methods for validation. PMID:26483609

  20. Stem Cell Tracking Technologies for Neurological Regenerative Medicine Purposes

    Yongtao Zheng

    2017-01-01

    Full Text Available The growing field of stem cell therapy is moving toward clinical trials in a variety of applications, particularly for neurological diseases. However, this translation of cell therapies into humans has prompted a need to create innovative and breakthrough methods for stem cell tracing, to explore the migration routes and its reciprocity with microenvironment targets in the body, to monitor and track the outcome after stem cell transplantation therapy, and to track the distribution and cell viability of transplanted cells noninvasively and longitudinally. Recently, a larger number of cell tracking methods in vivo were developed and applied in animals and humans, including magnetic resonance imaging, nuclear medicine imaging, and optical imaging. This review has been intended to summarize the current use of those imaging tools in tracking stem cells, detailing their main features and drawbacks, including image resolution, tissue penetrating depth, and biosafety aspects. Finally, we address that multimodality imaging method will be a more potential tracking tool in the future clinical application.

  1. Validation and application of Acoustic Mapping Velocimetry

    Baranya, Sandor; Muste, Marian

    2016-04-01

    The goal of this paper is to introduce a novel methodology to estimate bedload transport in rivers based on an improved bedform tracking procedure. The measurement technique combines components and processing protocols from two contemporary nonintrusive instruments: acoustic and image-based. The bedform mapping is conducted with acoustic surveys while the estimation of the velocity of the bedforms is obtained with processing techniques pertaining to image-based velocimetry. The technique is therefore called Acoustic Mapping Velocimetry (AMV). The implementation of this technique produces a whole-field velocity map associated with the multi-directional bedform movement. Based on the calculated two-dimensional bedform migration velocity field, the bedload transport estimation is done using the Exner equation. A proof-of-concept experiment was performed to validate the AMV based bedload estimation in a laboratory flume at IIHR-Hydroscience & Engineering (IIHR). The bedform migration was analysed at three different flow discharges. Repeated bed geometry mapping, using a multiple transducer array (MTA), provided acoustic maps, which were post-processed with a particle image velocimetry (PIV) method. Bedload transport rates were calculated along longitudinal sections using the streamwise components of the bedform velocity vectors and the measured bedform heights. The bulk transport rates were compared with the results from concurrent direct physical samplings and acceptable agreement was found. As a first field implementation of the AMV an attempt was made to estimate bedload transport for a section of the Ohio river in the United States, where bed geometry maps, resulted by repeated multibeam echo sounder (MBES) surveys, served as input data. Cross-sectional distributions of bedload transport rates from the AMV based method were compared with the ones obtained from another non-intrusive technique (due to the lack of direct samplings), ISSDOTv2, developed by the US Army

  2. Detecting and Tracking Nonfluorescent Nanoparticles Probes in Live Cells

    Wang, Gufeng; Fang, Ning

    2012-01-17

    Precisely imaging and tracking dynamic biological processes in live cells are crucial for both fundamental research in life sciences and biomedical applications. Nonfluorescent nanoparticles are emerging as important optical probes in live-cell imaging because of their excellent photostability, large optical cross sections, and low cytotoxicity. Here, we provide a review of recent development in optical imaging of nonfluorescent nanoparticle probes and their applications in dynamic tracking and biosensing in live cells. A brief discussion on cytotoxicity of nanoparticle probes is also provided.

  3. Dynamically constrained pipeline for tracking neural progenitor cells

    Vestergaard, Jacob Schack; Dahl, Anders; Holm, Peter

    2013-01-01

    . A mitosis detector constructed from empirical observations of cells in a pre-mitotic state interacts with the graph formulation to dynamically allow for cell mitosis when appropriate. Track consistency is ensured by introducing pragmatic constraints and the notion of blob states. We validate the proposed...

  4. Tracking plasma cell differentiation and survival.

    Roth, Katrin; Oehme, Laura; Zehentmeier, Sandra; Zhang, Yang; Niesner, Raluca; Hauser, Anja E

    2014-01-01

    Plasma cells play a crucial role for the humoral immune response as they represent the body's factories for antibody production. The differentiation from a B cell into a plasma cell is controlled by a complex transcriptional network and happens within secondary lymphoid organs. Based on their lifetime, two types of antibody secreting cells can be distinguished: Short-lived plasma cells are located in extrafollicular sites of secondary lymphoid organs such as lymph node medullary cords and the splenic red pulp. A fraction of plasmablasts migrate from secondary lymphoid organs to the bone marrow where they can become long-lived plasma cells. Bone marrow plasma cells reside in special microanatomical environments termed survival niches, which provide factors promoting their longevity. Reticular stromal cells producing the chemokine CXCL12, which is known to attract plasmablasts to the bone marrow but also to promote plasma cell survival, play a crucial role in the maintenance of these niches. In addition, hematopoietic cells are contributing to the niches by providing other soluble survival factors. Here, we review the current knowledge on the factors involved in plasma cell differentiation, their localization and migration. We also give an overview on what is known regarding the maintenance of long lived plasma cells in survival niches of the bone marrow. © 2013 International Society for Advancement of Cytometry.

  5. Full-field particle velocimetry with a photorefractive optical novelty filter

    Woerdemann, Mike; Holtmann, Frank; Denz, Cornelia

    2008-01-01

    We utilize the finite time constant of a photorefractive optical novelty filter microscope to access full-field velocity information of fluid flows on microscopic scales. In contrast to conventional methods such as particle image velocimetry and particle tracking velocimetry, not only image acquisition of the tracer particle field but also evaluation of tracer particle velocities is done all-optically by the novelty filter. We investigate the velocity dependent parameters of two-beam coupling based optical novelty filters and demonstrate calibration and application of a photorefractive velocimetry system. Theoretical and practical limits to the range of accessible velocities are discussed

  6. Track segment studies with Chinese hamster cells

    Bird, R.P.

    1984-01-01

    Survival curves of near-diploid and near-tetraploid Chinese hamster cell cultures following irradiation by an 241 Am α source indicate different growth rates for the two clones. Possible reasons for the difference are discussed

  7. TRACKING STEM CELLS IN AN INHERENTLY REGENRATIVE ENVIRONMENT

    Lauridsen, Henrik; Foldager, Casper Bindzus; Hagensen, Mette

    2012-01-01

    of such therapies. The objective of this study was to non-invasively evaluate regeneration over time in a truly regenerative process, the regeneration of an axolotl limb, employing superparamagnetic iron oxide particles (SPIO) contrast agents for stem cell tracking in MRI. Materials and Methods: Amputation of one...... in conjugation with the transfection agent poly-L-lysin (PLL) was tested on cultures of axolotl blastema cells from 7 animals in vitro. PicoGreen-DNA quantification following 3 weeks of culturing was performed to quantify cell viability. MRI-tracking of SPIO labelled blastema cells in the regenerating limb of 5....... Results: SPIO labelling with neither VSOP-C200, Resovist nor Resovist/PLL had any significant effect on blastema cell viability in vitro. Labelled tissue was clearly detectable in vivo 49 days after amputation using MRI (Fig. 1) and a significant decline in signal intensity of labelled limbs versus sham...

  8. Advective isotope transport by mixing cell and particle tracking algorithms

    Tezcan, L.; Meric, T.

    1999-01-01

    The 'mixing cell' algorithm of the environmental isotope data evaluation is integrated with the three dimensional finite difference ground water flow model (MODFLOW) to simulate the advective isotope transport and the approach is compared with the 'particle tracking' algorithm of the MOC3D, that simulates three-dimensional solute transport with the method of characteristics technique

  9. Tracking hypoxic signaling within encapsulated cell aggregates.

    Skiles, Matthew L; Sahai, Suchit; Blanchette, James O

    2011-12-16

    In Diabetes mellitus type 1, autoimmune destruction of the pancreatic β-cells results in loss of insulin production and potentially lethal hyperglycemia. As an alternative treatment option to exogenous insulin injection, transplantation of functional pancreatic tissue has been explored. This approach offers the promise of a more natural, long-term restoration of normoglycemia. Protection of the donor tissue from the host's immune system is required to prevent rejection and encapsulation is a method used to help achieve this aim. Biologically-derived materials, such as alginate and agarose, have been the traditional choice for capsule construction but may induce inflammation or fibrotic overgrowth which can impede nutrient and oxygen transport. Alternatively, synthetic poly(ethylene glycol) (PEG)-based hydrogels are non-degrading, easily functionalized, available at high purity, have controllable pore size, and are extremely biocompatible. As an additional benefit, PEG hydrogels may be formed rapidly in a simple photo-crosslinking reaction that does not require application of non-physiological temperatures. Such a procedure is described here. In the crosslinking reaction, UV degradation of the photoinitiator, 1-[4-(2-Hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propane-1-one (Irgacure 2959), produces free radicals which attack the vinyl carbon-carbon double bonds of dimethacrylated PEG (PEGDM) inducing crosslinking at the chain ends. Crosslinking can be achieved within 10 minutes. PEG hydrogels constructed in such a manner have been shown to favorably support cells, and the low photoinitiator concentration and brief exposure to UV irradiation is not detrimental to viability and function of the encapsulated tissue. While we methacrylate our PEG with the method described below, PEGDM can also be directly purchased from vendors such as Sigma. An inherent consequence of encapsulation is isolation of the cells from a vascular network. Supply of nutrients, notably oxygen

  10. Coaxial volumetric velocimetry

    Schneiders, Jan F. G.; Scarano, Fulvio; Jux, Constantin; Sciacchitano, Andrea

    2018-06-01

    This study describes the working principles of the coaxial volumetric velocimeter (CVV) for wind tunnel measurements. The measurement system is derived from the concept of tomographic PIV in combination with recent developments of Lagrangian particle tracking. The main characteristic of the CVV is its small tomographic aperture and the coaxial arrangement between the illumination and imaging directions. The system consists of a multi-camera arrangement subtending only few degrees solid angle and a long focal depth. Contrary to established PIV practice, laser illumination is provided along the same direction as that of the camera views, reducing the optical access requirements to a single viewing direction. The laser light is expanded to illuminate the full field of view of the cameras. Such illumination and imaging conditions along a deep measurement volume dictate the use of tracer particles with a large scattering area. In the present work, helium-filled soap bubbles are used. The fundamental principles of the CVV in terms of dynamic velocity and spatial range are discussed. Maximum particle image density is shown to limit tracer particle seeding concentration and instantaneous spatial resolution. Time-averaged flow fields can be obtained at high spatial resolution by ensemble averaging. The use of the CVV for time-averaged measurements is demonstrated in two wind tunnel experiments. After comparing the CVV measurements with the potential flow in front of a sphere, the near-surface flow around a complex wind tunnel model of a cyclist is measured. The measurements yield the volumetric time-averaged velocity and vorticity field. The measurements of the streamlines in proximity of the surface give an indication of the skin-friction lines pattern, which is of use in the interpretation of the surface flow topology.

  11. Schlieren image velocimetry measurements in a rocket engine exhaust plume

    Morales, Rudy; Peguero, Julio; Hargather, Michael

    2017-11-01

    Schlieren image velocimetry (SIV) measures velocity fields by tracking the motion of naturally-occurring turbulent flow features in a compressible flow. Here the technique is applied to measuring the exhaust velocity profile of a liquid rocket engine. The SIV measurements presented include discussion of visibility of structures, image pre-processing for structure visibility, and ability to process resulting images using commercial particle image velocimetry (PIV) codes. The small-scale liquid bipropellant rocket engine operates on nitrous oxide and ethanol as propellants. Predictions of the exhaust velocity are obtained through NASA CEA calculations and simple compressible flow relationships, which are compared against the measured SIV profiles. Analysis of shear layer turbulence along the exhaust plume edge is also presented.

  12. Optical fibre laser velocimetry: a review

    Charrett, Thomas O H; James, Stephen W; Tatam, Ralph P

    2012-01-01

    The applications of optical fibre technology to laser velocimetry are diverse and often critical to their successful implementation, particularly in harsh environments. Applications range from the use of optical fibres for beam delivery and scattered light collection, aiding the miniaturization of instrument probes, to the use of imaging fibre bundles for imaging the flow field in planar velocimetry systems. Optical fibre techniques have also been used in signal processing, for example fibre frequency shifters, and optical fibre devices such as amplifiers and lasers have been exploited. This paper will review the use of optical fibres in point-wise laser velocimetry techniques such as laser Doppler velocimetry and laser transit anemometry, as well as in planar measurement techniques such as particle imaging velocimetry and planar Doppler velocimetry. (topical review)

  13. Three-dimensional particle image velocimetry measurement technique

    Hassan, Y.A.; Seeley, C.H.; Henderson, J.A.; Schmidl, W.D.

    2004-01-01

    The experimental flow visualization tool, Particle Image Velocimetry (PIV), is being used to determine the velocity field in two-dimensional fluid flows. In the past few years, the technique has been improved to allow the capture of flow fields in three dimensions. This paper describes changes which were made to two existing two-dimensional tracking algorithms to enable them to track three-dimensional PIV data. Results of the tests performed on these three-dimensional routines with synthetic data are presented. Experimental data was also used to test the tracking algorithms. The test setup which was used to acquire the three-dimensional experimental data is described, along with the results from both of the tracking routines which were used to analyze the experimental data. (author)

  14. A track-event theory of cell survival

    Besserer, Juergen; Schneider, Uwe

    2015-01-01

    When fractionation schemes for hypofractionation and stereotactic body radiotherapy are considered, a reliable cell survival model at high dose is needed for calculating doses of similar biological effectiveness. In this work a simple model for cell survival which is valid also at high dose is developed from Poisson statistics. An event is defined by two double strand breaks (DSB) on the same or different chromosomes. An event is always lethal due to direct lethal damage or lethal binary misrepair by the formation of chromosome aberrations. Two different mechanisms can produce events: one-track events (OTE) or two-track-events (TTE). The target for an OTE is always a lethal event, the target for an TTE is one DSB. At least two TTEs on the same or different chromosomes are necessary to produce an event. Both, the OTE and the TTE are statistically independent. From the stochastic nature of cell kill which is described by the Poisson distribution the cell survival probability was derived. It was shown that a solution based on Poisson statistics exists for cell survival. It exhibits exponential cell survival at high dose and a finite gradient of cell survival at vanishing dose, which is in agreement with experimental cell studies. The model fits the experimental data nearly as well as the three-parameter formula of Hug-Kellerer and is only based on two free parameters. It is shown that the LQ formalism is an approximation of the model derived in this work. It could be also shown that the derived model predicts a fractionated cell survival experiment better than the LQ-model. It was shown that cell survival can be described with a simple analytical formula on the basis of Poisson statistics. This solution represents in the limit of large dose the typical exponential behavior and predicts cell survival after fractionated dose application better than the LQ-model.

  15. A track-event theory of cell survival

    Besserer, Juergen; Schneider, Uwe [Zuerich Univ. (Switzerland). Inst. of Physics; Radiotherapy Hirslanden, Zuerich (Switzerland)

    2015-09-01

    When fractionation schemes for hypofractionation and stereotactic body radiotherapy are considered, a reliable cell survival model at high dose is needed for calculating doses of similar biological effectiveness. In this work a simple model for cell survival which is valid also at high dose is developed from Poisson statistics. An event is defined by two double strand breaks (DSB) on the same or different chromosomes. An event is always lethal due to direct lethal damage or lethal binary misrepair by the formation of chromosome aberrations. Two different mechanisms can produce events: one-track events (OTE) or two-track-events (TTE). The target for an OTE is always a lethal event, the target for an TTE is one DSB. At least two TTEs on the same or different chromosomes are necessary to produce an event. Both, the OTE and the TTE are statistically independent. From the stochastic nature of cell kill which is described by the Poisson distribution the cell survival probability was derived. It was shown that a solution based on Poisson statistics exists for cell survival. It exhibits exponential cell survival at high dose and a finite gradient of cell survival at vanishing dose, which is in agreement with experimental cell studies. The model fits the experimental data nearly as well as the three-parameter formula of Hug-Kellerer and is only based on two free parameters. It is shown that the LQ formalism is an approximation of the model derived in this work. It could be also shown that the derived model predicts a fractionated cell survival experiment better than the LQ-model. It was shown that cell survival can be described with a simple analytical formula on the basis of Poisson statistics. This solution represents in the limit of large dose the typical exponential behavior and predicts cell survival after fractionated dose application better than the LQ-model.

  16. Untangling cell tracks: Quantifying cell migration by time lapse image data analysis.

    Svensson, Carl-Magnus; Medyukhina, Anna; Belyaev, Ivan; Al-Zaben, Naim; Figge, Marc Thilo

    2018-03-01

    Automated microscopy has given researchers access to great amounts of live cell imaging data from in vitro and in vivo experiments. Much focus has been put on extracting cell tracks from such data using a plethora of segmentation and tracking algorithms, but further analysis is normally required to draw biologically relevant conclusions. Such relevant conclusions may be whether the migration is directed or not, whether the population has homogeneous or heterogeneous migration patterns. This review focuses on the analysis of cell migration data that are extracted from time lapse images. We discuss a range of measures and models used to analyze cell tracks independent of the biological system or the way the tracks were obtained. For single-cell migration, we focus on measures and models giving examples of biological systems where they have been applied, for example, migration of bacteria, fibroblasts, and immune cells. For collective migration, we describe the model systems wound healing, neural crest migration, and Drosophila gastrulation and discuss methods for cell migration within these systems. We also discuss the role of the extracellular matrix and subsequent differences between track analysis in vitro and in vivo. Besides methods and measures, we are putting special focus on the need for openly available data and code, as well as a lack of common vocabulary in cell track analysis. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  17. Accurately tracking single-cell movement trajectories in microfluidic cell sorting devices.

    Jeong, Jenny; Frohberg, Nicholas J; Zhou, Enlu; Sulchek, Todd; Qiu, Peng

    2018-01-01

    Microfluidics are routinely used to study cellular properties, including the efficient quantification of single-cell biomechanics and label-free cell sorting based on the biomechanical properties, such as elasticity, viscosity, stiffness, and adhesion. Both quantification and sorting applications require optimal design of the microfluidic devices and mathematical modeling of the interactions between cells, fluid, and the channel of the device. As a first step toward building such a mathematical model, we collected video recordings of cells moving through a ridged microfluidic channel designed to compress and redirect cells according to cell biomechanics. We developed an efficient algorithm that automatically and accurately tracked the cell trajectories in the recordings. We tested the algorithm on recordings of cells with different stiffness, and showed the correlation between cell stiffness and the tracked trajectories. Moreover, the tracking algorithm successfully picked up subtle differences of cell motion when passing through consecutive ridges. The algorithm for accurately tracking cell trajectories paves the way for future efforts of modeling the flow, forces, and dynamics of cell properties in microfluidics applications.

  18. Accurately tracking single-cell movement trajectories in microfluidic cell sorting devices.

    Jenny Jeong

    Full Text Available Microfluidics are routinely used to study cellular properties, including the efficient quantification of single-cell biomechanics and label-free cell sorting based on the biomechanical properties, such as elasticity, viscosity, stiffness, and adhesion. Both quantification and sorting applications require optimal design of the microfluidic devices and mathematical modeling of the interactions between cells, fluid, and the channel of the device. As a first step toward building such a mathematical model, we collected video recordings of cells moving through a ridged microfluidic channel designed to compress and redirect cells according to cell biomechanics. We developed an efficient algorithm that automatically and accurately tracked the cell trajectories in the recordings. We tested the algorithm on recordings of cells with different stiffness, and showed the correlation between cell stiffness and the tracked trajectories. Moreover, the tracking algorithm successfully picked up subtle differences of cell motion when passing through consecutive ridges. The algorithm for accurately tracking cell trajectories paves the way for future efforts of modeling the flow, forces, and dynamics of cell properties in microfluidics applications.

  19. Tracking the mechanical dynamics of human embryonic stem cell chromatin

    Hinde Elizabeth

    2012-12-01

    Full Text Available Abstract Background A plastic chromatin structure has emerged as fundamental to the self-renewal and pluripotent capacity of embryonic stem (ES cells. Direct measurement of chromatin dynamics in vivo is, however, challenging as high spatiotemporal resolution is required. Here, we present a new tracking-based method which can detect high frequency chromatin movement and quantify the mechanical dynamics of chromatin in live cells. Results We use this method to study how the mechanical properties of chromatin movement in human embryonic stem cells (hESCs are modulated spatiotemporally during differentiation into cardiomyocytes (CM. Notably, we find that pluripotency is associated with a highly discrete, energy-dependent frequency of chromatin movement that we refer to as a ‘breathing’ state. We find that this ‘breathing’ state is strictly dependent on the metabolic state of the cell and is progressively silenced during differentiation. Conclusions We thus propose that the measured chromatin high frequency movements in hESCs may represent a hallmark of pluripotency and serve as a mechanism to maintain the genome in a transcriptionally accessible state. This is a result that could not have been observed without the high spatial and temporal resolution provided by this novel tracking method.

  20. Large scale tracking of stem cells using sparse coding and coupled graphs

    Vestergaard, Jacob Schack; Dahl, Anders Lindbjerg; Holm, Peter

    Stem cell tracking is an inherently large scale problem. The challenge is to identify and track hundreds or thousands of cells over a time period of several weeks. This requires robust methods that can leverage the knowledge of specialists on the field. The tracking pipeline presented here consists...

  1. Uptake of magnetic nanoparticles into cells for cell tracking

    Becker, Christiane; Hodenius, Michael; Blendinger, Gitta; Sechi, Antonio; Hieronymus, Thomas; Mueller-Schulte, Detlef; Schmitz-Rode, Thomas; Zenke, Martin

    2007-01-01

    A challenge for future applications in nanotechnology is the functional integration of nano-sized materials into cellular structures. Here we investigated superparamagnetic Fe 3 O 4 iron oxide nanoparticles coated with a lipid bilayer for uptake into cells and for targeting subcellular compartments. It was found that magnetic nanoparticles (MNPs) are effectively taken up into cells and make cells acquire magnetic activity. Biotin-conjugated MNPs were further functionalized by binding of the fluorescent tag streptavidin-fluorescein isothiocyanate (FITC) and, following uptake into cells, shown to confer magnetic activity and fluorescence labeling. Such FITC-MNPs were localized in the lysosomal compartment of cells which suggests a receptor-mediated uptake mechanism

  2. Volumetric velocimetry for fluid flows

    Discetti, Stefano; Coletti, Filippo

    2018-04-01

    In recent years, several techniques have been introduced that are capable of extracting 3D three-component velocity fields in fluid flows. Fast-paced developments in both hardware and processing algorithms have generated a diverse set of methods, with a growing range of applications in flow diagnostics. This has been further enriched by the increasingly marked trend of hybridization, in which the differences between techniques are fading. In this review, we carry out a survey of the prominent methods, including optical techniques and approaches based on medical imaging. An overview of each is given with an example of an application from the literature, while focusing on their respective strengths and challenges. A framework for the evaluation of velocimetry performance in terms of dynamic spatial range is discussed, along with technological trends and emerging strategies to exploit 3D data. While critical challenges still exist, these observations highlight how volumetric techniques are transforming experimental fluid mechanics, and that the possibilities they offer have just begun to be explored.

  3. Continuous and simultaneous measurement of the tank-treading motion of red blood cells and the surrounding flow using translational confocal micro-particle image velocimetry (micro-PIV) with sub-micron resolution

    Oishi, M; Utsubo, K; Kinoshita, H; Fujii, T; Oshima, M

    2012-01-01

    In this study, a translational confocal micro-particle image velocimetry (PIV) system is introduced to measure the microscopic interaction between red blood cells (RBCs) and the surrounding flow. Since the macroscopic behavior of RBCs, such as the tank-treading motion, is closely related to the axial migration and other flow characteristics in arterioles, the measurement method must answer the conflicting demands of sub-micron resolution, continuous measurement and applicability for high-speed flow. In order to avoid loss of the measurement target, i.e. RBCs, from the narrow field of view during high-magnification measurement, the translation stage with the flow device moves in the direction opposite the direction of flow. The proposed system achieves the measurement of higher absolute velocities compared with a conventional confocal micro-PIV system without the drawbacks derived from stage vibration. In addition, we have applied a multicolor separation unit, which can measure different phases simultaneously using different fluorescent particles, in order to clarify the interaction between RBCs and the surrounding flow. Based on our measurements, the tank-treading motion of RBCs depends on the shear stress gradient of the surrounding flow. Although, the relationship between the tank-treading frequency and the shear rate of the surrounding flow is of the same order as in the previous uniform shear rate experiments, our results reveal the remarkable behavior of the non-uniform membrane velocities and lateral velocity component of flow around the RBCs. (paper)

  4. Robust cell tracking in epithelial tissues through identification of maximum common subgraphs.

    Kursawe, Jochen; Bardenet, Rémi; Zartman, Jeremiah J; Baker, Ruth E; Fletcher, Alexander G

    2016-11-01

    Tracking of cells in live-imaging microscopy videos of epithelial sheets is a powerful tool for investigating fundamental processes in embryonic development. Characterizing cell growth, proliferation, intercalation and apoptosis in epithelia helps us to understand how morphogenetic processes such as tissue invagination and extension are locally regulated and controlled. Accurate cell tracking requires correctly resolving cells entering or leaving the field of view between frames, cell neighbour exchanges, cell removals and cell divisions. However, current tracking methods for epithelial sheets are not robust to large morphogenetic deformations and require significant manual interventions. Here, we present a novel algorithm for epithelial cell tracking, exploiting the graph-theoretic concept of a 'maximum common subgraph' to track cells between frames of a video. Our algorithm does not require the adjustment of tissue-specific parameters, and scales in sub-quadratic time with tissue size. It does not rely on precise positional information, permitting large cell movements between frames and enabling tracking in datasets acquired at low temporal resolution due to experimental constraints such as phototoxicity. To demonstrate the method, we perform tracking on the Drosophila embryonic epidermis and compare cell-cell rearrangements to previous studies in other tissues. Our implementation is open source and generally applicable to epithelial tissues. © 2016 The Authors.

  5. Cross-correlation Doppler global velocimetry (CC-DGV)

    Cadel, Daniel R.; Lowe, K. Todd

    2015-08-01

    A flow velocimetry method, cross-correlation Doppler global velocimetry (CC-DGV), is presented as a robust, simplified, and high dynamic range implementation of the Doppler global/planar Doppler velocimetry technique. A sweep of several gigahertz of the vapor absorption spectrum is used for each velocity sample, with signals acquired from both Doppler-shifted scattered light within the flow and a non-Doppler shifted reference beam. Cross-correlation of these signals yields the Doppler shift between them, averaged over the duration of the scan. With presently available equipment, velocities from 0 ms-1 to over 3000 ms-1 can notionally be measured simultaneously, making the technique ideal for high speed flows. The processing routine is shown to be robust against large changes in the vapor pressure of the iodine cell, benefiting performance of the system in facilities where ambient conditions cannot be easily regulated. Validation of the system was performed with measurements of a model wind turbine blade boundary layer made in a 1.83 m by 1.83 m subsonic wind tunnel for which laser Doppler velocimetry (LDV) measurements were acquired alongside the CC-DGV results. CC-DGV uncertainties of ±1.30 ms-1, ±0.64 ms-1, and ±1.11 ms-1 were determined for the orthogonal stream-wise, transverse-horizontal, and transverse-vertical velocity components, and root-mean-square deviations of 2.77 ms-1 and 1.34 ms-1 from the LDV validation results were observed for Reynolds numbers of 1.5 million and 2 million, respectively. Volumetric mean velocity measurements are also presented for a supersonic jet, with velocity uncertainties of ±4.48 ms-1, ±16.93 ms-1, and ±0.50 ms-1 for the orthogonal components, and self-validation done by collapsing the data with a physical scaling.

  6. A novel validation algorithm allows for automated cell tracking and the extraction of biologically meaningful parameters.

    Daniel H Rapoport

    Full Text Available Automated microscopy is currently the only method to non-invasively and label-free observe complex multi-cellular processes, such as cell migration, cell cycle, and cell differentiation. Extracting biological information from a time-series of micrographs requires each cell to be recognized and followed through sequential microscopic snapshots. Although recent attempts to automatize this process resulted in ever improving cell detection rates, manual identification of identical cells is still the most reliable technique. However, its tedious and subjective nature prevented tracking from becoming a standardized tool for the investigation of cell cultures. Here, we present a novel method to accomplish automated cell tracking with a reliability comparable to manual tracking. Previously, automated cell tracking could not rival the reliability of manual tracking because, in contrast to the human way of solving this task, none of the algorithms had an independent quality control mechanism; they missed validation. Thus, instead of trying to improve the cell detection or tracking rates, we proceeded from the idea to automatically inspect the tracking results and accept only those of high trustworthiness, while rejecting all other results. This validation algorithm works independently of the quality of cell detection and tracking through a systematic search for tracking errors. It is based only on very general assumptions about the spatiotemporal contiguity of cell paths. While traditional tracking often aims to yield genealogic information about single cells, the natural outcome of a validated cell tracking algorithm turns out to be a set of complete, but often unconnected cell paths, i.e. records of cells from mitosis to mitosis. This is a consequence of the fact that the validation algorithm takes complete paths as the unit of rejection/acceptance. The resulting set of complete paths can be used to automatically extract important biological parameters

  7. System for tracking transplanted limbal epithelial stem cells in the treatment of corneal stem cell deficiency

    Boadi, J.; Sangwal, V.; MacNeil, S.; Matcher, S. J.

    2015-03-01

    The prevailing hypothesis for the existence and healing of the avascular corneal epithelium is that this layer of cells is continually produced by stem cells in the limbus and transported onto the cornea to mature into corneal epithelium. Limbal Stem Cell Deficiency (LSCD), in which the stem cell population is depleted, can lead to blindness. LSCD can be caused by chemical and thermal burns to the eye. A popular treatment, especially in emerging economies such as India, is the transplantation of limbal stem cells onto damaged limbus with hope of repopulating the region. Hence regenerating the corneal epithelium. In order to gain insights into the success rates of this treatment, new imaging technologies are needed in order to track the transplanted cells. Optical Coherence Tomography (OCT) is well known for its high resolution in vivo images of the retina. A custom OCT system has been built to image the corneal surface, to investigate the fate of transplanted limbal stem cells. We evaluate two methods to label and track transplanted cells: melanin labelling and magneto-labelling. To evaluate melanin labelling, stem cells are loaded with melanin and then transplanted onto a rabbit cornea denuded of its epithelium. The melanin displays strongly enhanced backscatter relative to normal cells. To evaluate magneto-labelling the stem cells are loaded with magnetic nanoparticles (20-30nm in size) and then imaged with a custom-built, magneto-motive OCT system.

  8. Single-particle tracking of quantum dot-conjugated prion proteins inside yeast cells

    Tsuji, Toshikazu; Kawai-Noma, Shigeko [Department of Biomolecular Engineering, Graduate School of Biosciences and Biotechnology, Tokyo Institute of Technology, B56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501 (Japan); Pack, Chan-Gi [Cellular Informatics Laboratory, RIKEN Advanced Science Institute, Wako-shi, Saitama 351-0198 (Japan); Terajima, Hideki [Department of Biomolecular Engineering, Graduate School of Biosciences and Biotechnology, Tokyo Institute of Technology, B56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501 (Japan); Yajima, Junichiro; Nishizaka, Takayuki [Department of Physics, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588 (Japan); Kinjo, Masataka [Laboratory of Molecular Cell Dynamics, Graduate School of Life Sciences, Hokkaido University, Sapporo 001-0021 (Japan); Taguchi, Hideki, E-mail: taguchi@bio.titech.ac.jp [Department of Biomolecular Engineering, Graduate School of Biosciences and Biotechnology, Tokyo Institute of Technology, B56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501 (Japan)

    2011-02-25

    Research highlights: {yields} We develop a method to track a quantum dot-conjugated protein in yeast cells. {yields} We incorporate the conjugated quantum dot proteins into yeast spheroplasts. {yields} We track the motions by conventional or 3D tracking microscopy. -- Abstract: Yeast is a model eukaryote with a variety of biological resources. Here we developed a method to track a quantum dot (QD)-conjugated protein in the budding yeast Saccharomyces cerevisiae. We chemically conjugated QDs with the yeast prion Sup35, incorporated them into yeast spheroplasts, and tracked the motions by conventional two-dimensional or three-dimensional tracking microscopy. The method paves the way toward the individual tracking of proteins of interest inside living yeast cells.

  9. Single-particle tracking of quantum dot-conjugated prion proteins inside yeast cells

    Tsuji, Toshikazu; Kawai-Noma, Shigeko; Pack, Chan-Gi; Terajima, Hideki; Yajima, Junichiro; Nishizaka, Takayuki; Kinjo, Masataka; Taguchi, Hideki

    2011-01-01

    Research highlights: → We develop a method to track a quantum dot-conjugated protein in yeast cells. → We incorporate the conjugated quantum dot proteins into yeast spheroplasts. → We track the motions by conventional or 3D tracking microscopy. -- Abstract: Yeast is a model eukaryote with a variety of biological resources. Here we developed a method to track a quantum dot (QD)-conjugated protein in the budding yeast Saccharomyces cerevisiae. We chemically conjugated QDs with the yeast prion Sup35, incorporated them into yeast spheroplasts, and tracked the motions by conventional two-dimensional or three-dimensional tracking microscopy. The method paves the way toward the individual tracking of proteins of interest inside living yeast cells.

  10. Tracking single cells in live animals using a photoconvertible near-infrared cell membrane label.

    Carlson, Alicia L; Fujisaki, Joji; Wu, Juwell; Runnels, Judith M; Turcotte, Raphaël; Spencer, Joel A; Celso, Cristina Lo; Scadden, David T; Strom, Terry B; Lin, Charles P

    2013-01-01

    We describe a novel photoconversion technique to track individual cells in vivo using a commercial lipophilic membrane dye, DiR. We show that DiR exhibits a permanent fluorescence emission shift (photoconversion) after light exposure and does not reacquire the original color over time. Ratiometric imaging can be used to distinguish photoconverted from non-converted cells with high sensitivity. Combining the use of this photoconvertible dye with intravital microscopy, we tracked the division of individual hematopoietic stem/progenitor cells within the calvarium bone marrow of live mice. We also studied the peripheral differentiation of individual T cells by tracking the gain or loss of FoxP3-GFP expression, a marker of the immune suppressive function of CD4(+) T cells. With the near-infrared photoconvertible membrane dye, the entire visible spectral range is available for simultaneous use with other fluorescent proteins to monitor gene expression or to trace cell lineage commitment in vivo with high spatial and temporal resolution.

  11. Development of Hydroxyl Tagging Velocimetry for Low Velocity Flows

    Andre, Matthieu A.; Bardet, Philippe M.; Burns, Ross A.; Danehy, Paul M.

    2016-01-01

    Hydroxyl tagging velocimetry (HTV) is a molecular tagging technique that relies on the photo-dissociation of water vapor into OH radicals and their subsequent tracking using laser induced fluorescence. Velocities are then obtained from time-of-flight calculations. At ambient temperature in air, the OH species lifetime is relatively short (<50 µs), making it suited for high speed flows. Lifetime and radicals formation increases with temperature, which allows HTV to also probe low-velocity, high-temperature flows or reacting flows such as flames. The present work aims at extending the domain of applicability of HTV, particularly towards low-speed (<10 m/s) and moderate (<500 K) temperature flows. Results are compared to particle image velocimetry (PIV) measurements recorded in identical conditions. Single shot and averaged velocity profiles are obtained in an air jet at room temperature. By modestly raising the temperature (100-200 degC) the OH production increases, resulting in an improvement of the signal-to-noise ratio (SNR). Use of nitrogen - a non-reactive gas with minimal collisional quenching - extends the OH species lifetime (to over 500 µs), which allows probing of slower flows or, alternately, increases the measurement precision at the expense of spatial resolution. Instantaneous velocity profiles are resolved in a 100degC nitrogen jet (maximum jet-center velocity of 6.5 m/s) with an uncertainty down to 0.10 m/s (1.5%) at 68% confidence level. MTV measurements are compared with particle image velocimetry and show agreement within 2%.

  12. Advanced cell therapies: targeting, tracking and actuation of cells with magnetic particles.

    Connell, John J; Patrick, P Stephen; Yu, Yichao; Lythgoe, Mark F; Kalber, Tammy L

    2015-01-01

    Regenerative medicine would greatly benefit from a new platform technology that enabled measurable, controllable and targeting of stem cells to a site of disease or injury in the body. Superparamagnetic iron-oxide nanoparticles offer attractive possibilities in biomedicine and can be incorporated into cells, affording a safe and reliable means of tagging. This review describes three current and emerging methods to enhance regenerative medicine using magnetic particles to guide therapeutic cells to a target organ; track the cells using MRI and assess their spatial localization with high precision and influence the behavior of the cell using magnetic actuation. This approach is complementary to the systemic injection of cell therapies, thus expanding the horizon of stem cell therapeutics.

  13. [Study on method of tracking the active cells in image sequences based on EKF-PF].

    Tang, Chunming; Liu, Ying

    2013-02-01

    In cell image sequences, due to the nonlinear and nonGaussian motion characteristics of active cells, the accurate prediction and tracking is still an unsolved problem. We applied extended Kalman particle filter (EKF-PF) here in our study, attempting to solve the problem. Firstly we confirmed the existence and positions of the active cells. Then we established a motion model and improved it via adding motion angle estimation. Next we predicted motion parameters, such as displacement, velocity, accelerated velocity and motion angle, in region centers of the cells being tracked. Finally we obtained the motion traces of active cells. There were fourteen active cells in three image sequences which have been tracked. The errors were less than 2.5 pixels when the prediction values were compared with actual values. It showed that the presented algorithm may basically reach the solution of accurate predition and tracking of the active cells.

  14. Automated cell tracking and analysis in phase-contrast videos (iTrack4U): development of Java software based on combined mean-shift processes.

    Cordelières, Fabrice P; Petit, Valérie; Kumasaka, Mayuko; Debeir, Olivier; Letort, Véronique; Gallagher, Stuart J; Larue, Lionel

    2013-01-01

    Cell migration is a key biological process with a role in both physiological and pathological conditions. Locomotion of cells during embryonic development is essential for their correct positioning in the organism; immune cells have to migrate and circulate in response to injury. Failure of cells to migrate or an inappropriate acquisition of migratory capacities can result in severe defects such as altered pigmentation, skull and limb abnormalities during development, and defective wound repair, immunosuppression or tumor dissemination. The ability to accurately analyze and quantify cell migration is important for our understanding of development, homeostasis and disease. In vitro cell tracking experiments, using primary or established cell cultures, are often used to study migration as cells can quickly and easily be genetically or chemically manipulated. Images of the cells are acquired at regular time intervals over several hours using microscopes equipped with CCD camera. The locations (x,y,t) of each cell on the recorded sequence of frames then need to be tracked. Manual computer-assisted tracking is the traditional method for analyzing the migratory behavior of cells. However, this processing is extremely tedious and time-consuming. Most existing tracking algorithms require experience in programming languages that are unfamiliar to most biologists. We therefore developed an automated cell tracking program, written in Java, which uses a mean-shift algorithm and ImageJ as a library. iTrack4U is a user-friendly software. Compared to manual tracking, it saves considerable amount of time to generate and analyze the variables characterizing cell migration, since they are automatically computed with iTrack4U. Another major interest of iTrack4U is the standardization and the lack of inter-experimenter differences. Finally, iTrack4U is adapted for phase contrast and fluorescent cells.

  15. Automated cell tracking and analysis in phase-contrast videos (iTrack4U: development of Java software based on combined mean-shift processes.

    Fabrice P Cordelières

    Full Text Available Cell migration is a key biological process with a role in both physiological and pathological conditions. Locomotion of cells during embryonic development is essential for their correct positioning in the organism; immune cells have to migrate and circulate in response to injury. Failure of cells to migrate or an inappropriate acquisition of migratory capacities can result in severe defects such as altered pigmentation, skull and limb abnormalities during development, and defective wound repair, immunosuppression or tumor dissemination. The ability to accurately analyze and quantify cell migration is important for our understanding of development, homeostasis and disease. In vitro cell tracking experiments, using primary or established cell cultures, are often used to study migration as cells can quickly and easily be genetically or chemically manipulated. Images of the cells are acquired at regular time intervals over several hours using microscopes equipped with CCD camera. The locations (x,y,t of each cell on the recorded sequence of frames then need to be tracked. Manual computer-assisted tracking is the traditional method for analyzing the migratory behavior of cells. However, this processing is extremely tedious and time-consuming. Most existing tracking algorithms require experience in programming languages that are unfamiliar to most biologists. We therefore developed an automated cell tracking program, written in Java, which uses a mean-shift algorithm and ImageJ as a library. iTrack4U is a user-friendly software. Compared to manual tracking, it saves considerable amount of time to generate and analyze the variables characterizing cell migration, since they are automatically computed with iTrack4U. Another major interest of iTrack4U is the standardization and the lack of inter-experimenter differences. Finally, iTrack4U is adapted for phase contrast and fluorescent cells.

  16. The use of cell phone network data in traffic data collection and long-haul truckshed (geographic extent) tracking.

    2012-12-01

    This study analyzed the potential of cell phone positioning techniques in freight truck data collection and long-haul : truckshed (geographic extent) tracking. Freight truck identification and tracking algorithms were developed by means of : cell pho...

  17. Iron Administration before Stem Cell Harvest Enables MR Imaging Tracking after Transplantation

    Khurana, Aman; Chapelin, Fanny; Beck, Graham; Lenkov, Olga D.; Donig, Jessica; Nejadnik, Hossein; Messing, Solomon; Derugin, Nikita; Chan, Ray Chun-Fai; Gaur, Amitabh; Sennino, Barbara; McDonald, Donald M.; Kempen, Paul J.; Tikhomirov, Grigory A.; Rao, Jianghong

    2013-01-01

    Transplanted mesenchymal stem cells (MSCs) could be detected and tracked with MR imaging, if the donor is treated with an intravenous injection of the Food and Drug Administration–approved iron supplement ferumoxytol prior to MSC harvesting.

  18. Iron Oxide as an Mri Contrast Agent for Cell Tracking: Supplementary Issue

    Daniel J. Korchinski

    2015-01-01

    Full Text Available Iron oxide contrast agents have been combined with magnetic resonance imaging for cell tracking. In this review, we discuss coating properties and provide an overview of ex vivo and in vivo labeling of different cell types, including stem cells, red blood cells, and monocytes/macrophages. Furthermore, we provide examples of applications of cell tracking with iron contrast agents in stroke, multiple sclerosis, cancer, arteriovenous malformations, and aortic and cerebral aneurysms. Attempts at quantifying iron oxide concentrations and other vascular properties are examined. We advise on designing studies using iron contrast agents including methods for validation.

  19. In vitro detection of circulating tumor cells compared by the CytoTrack and CellSearch methods

    Hillig, T.; Horn, P.; Nygaard, Ann-Britt

    2015-01-01

    .23/p = 0.09). Overall, the recovery of CytoTrack and CellSearch was 68.8 +/- 3.9 %/71.1 +/- 2.9 %, respectively (p = 0.58). In spite of different methodologies, CytoTrack and CellSearch found similar number of CTCs, when spiking was performed with the EpCAM and pan cytokeratin-positive cell line MCF-7......Comparison of two methods to detect circulating tumor cells (CTC) CytoTrack and CellSearch through recovery of MCF-7 breast cancer cells, spiked into blood collected from healthy donors. Spiking of a fixed number of EpCAM and pan-cytokeratin positive MCF-7 cells into 7.5 mL donor blood...... was performed by FACSAria flow sorting. The samples were shipped to either CytoTrack or CellSearch research facilities within 48 h, where evaluation of MCF-7 recovery was performed. CytoTrack and CellSearch analyses were performed simultaneously. Recoveries of MCF-7 single cells, cells in clusters, and clusters...

  20. Simultaneous cell tracking and image alignment in 3D CLSM imagery of growing arabidopsis thaliana sepals

    Fick, R.H.J.; Fedorov, D.; Roeder, A.H.K.; Manjunath, B.S.

    2013-01-01

    In this research we propose a combined cell matching and image alignment method for tracking cells based on their nuclear locations in 3D fluorescent Confocal Laser Scanning Microscopy (CLSM) image sequences. We then apply it to study the cell division pattern in the developing sepal of the small

  1. Use of Nanoparticle Contrast Agents for Cell Tracking with Computed Tomography

    2017-01-01

    Efforts to develop novel cell-based therapies originated with the first bone marrow transplant on a leukemia patient in 1956. Preclinical and clinical examples of cell-based treatment strategies have shown promising results across many disciplines in medicine, with recent advances in immune cell therapies for cancer producing remarkable response rates, even in patients with multiple treatment failures. However, cell-based therapies suffer from inconsistent outcomes, motivating the search for tools that allow monitoring of cell delivery and behavior in vivo. Noninvasive cell imaging techniques, also known as cell tracking, have been developed to address this issue. These tools can allow real-time, quantitative, and long-term monitoring of transplanted cells in the recipient, providing insight on cell migration, distribution, viability, differentiation, and fate, all of which play crucial roles in treatment efficacy. Understanding these parameters allows the optimization of cell choice, delivery route, and dosage for therapy and advances cell-based therapy for specific clinical uses. To date, most cell tracking work has centered on imaging modalities such as MRI, radionuclide imaging, and optical imaging. However, X-ray computed tomography (CT) is an emerging method for cell tracking that has several strengths such as high spatial and temporal resolution, and excellent quantitative capabilities. The advantages of CT for cell tracking are enhanced by its wide availability and cost effectiveness, allowing CT to become one of the most popular clinical imaging modalities and a key asset in disease diagnosis. In this review, we will discuss recent advances in cell tracking methods using X-ray CT in various applications, in addition to predictions on how the field will progress. PMID:28485976

  2. Robust Individual-Cell/Object Tracking via PCANet Deep Network in Biomedicine and Computer Vision

    Bineng Zhong

    2016-01-01

    Full Text Available Tracking individual-cell/object over time is important in understanding drug treatment effects on cancer cells and video surveillance. A fundamental problem of individual-cell/object tracking is to simultaneously address the cell/object appearance variations caused by intrinsic and extrinsic factors. In this paper, inspired by the architecture of deep learning, we propose a robust feature learning method for constructing discriminative appearance models without large-scale pretraining. Specifically, in the initial frames, an unsupervised method is firstly used to learn the abstract feature of a target by exploiting both classic principal component analysis (PCA algorithms with recent deep learning representation architectures. We use learned PCA eigenvectors as filters and develop a novel algorithm to represent a target by composing of a PCA-based filter bank layer, a nonlinear layer, and a patch-based pooling layer, respectively. Then, based on the feature representation, a neural network with one hidden layer is trained in a supervised mode to construct a discriminative appearance model. Finally, to alleviate the tracker drifting problem, a sample update scheme is carefully designed to keep track of the most representative and diverse samples during tracking. We test the proposed tracking method on two standard individual cell/object tracking benchmarks to show our tracker's state-of-the-art performance.

  3. Application of particle imaging velocimetry in windtunnels

    Kompenhans, J.; Reichmuth, J.

    1987-01-01

    Recently the instantaneous and nonintrusive measurement of the flow velocity in a large area of the flow field (two-dimensional plane) became possible by means of particle imaging velocimetry (PIV). Up to now PIV has mainly been used for model experiments at low flow velocities in order to test and to improve the measuring technique. The present aim is the application of PIV in large wind tunnels at high flow velocities. 7 references

  4. Velocimetry Overview for visitors from the DOD

    Briggs, Matthew E. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Physics Division; Holtkamp, David Bruce [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). Physics Division

    2016-08-19

    We are in the midst of a transformative period in which technological advances are making fundamental changes in the measurement techniques that form the backbone of nuclear weapon certification. Optical velocimetry has replaced electrical shorting pins in “Hydrotests,” which measure the dynamic implosion process. This advance has revolutionized nuclear weapons certification during the last 5 years. We can now measure the implosion process that drives a nuclear detonation with many orders of magnitude more resolution in both space and time than was possible just 10 years ago. It has been compared to going from Morse Code to HDTV, resulting in a dozen or more improvements in models of these weapons. These Hydrotests are carried out at LANL, LLNL and the NNSS, with the later holding the important role of allowing us to test with nuclear materials, in sub-critical configurations (i.e., no yield.) Each of these institutions has largely replaced pins with hundreds of channels of optical velocimetry. Velocimetry is non-contact and is used simultaneously with the X-ray capability of these facilities. The U1-a facility at NNSS pioneered this approach in the Gemini series in 2012, and continues to lead, both in channel count and technological advances. Close cooperation among LANL, LLNL and NSTec in these advances serves the complex by leveraging capabilities across sites and accelerating the pace of technical improvements.

  5. Convolutional Deep Belief Networks for Single-Cell/Object Tracking in Computational Biology and Computer Vision

    Zhong, Bineng; Pan, Shengnan; Zhang, Hongbo; Wang, Tian; Du, Jixiang; Chen, Duansheng; Cao, Liujuan

    2016-01-01

    In this paper, we propose deep architecture to dynamically learn the most discriminative features from data for both single-cell and object tracking in computational biology and computer vision. Firstly, the discriminative features are automatically learned via a convolutional deep belief network (CDBN). Secondly, we design a simple yet effective method to transfer features learned from CDBNs on the source tasks for generic purpose to the object tracking tasks using only limited amount of tra...

  6. Cell survival in carbon beams - comparison of amorphous track model predictions

    Grzanka, L.; Greilich, S.; Korcyl, M.

    Introduction: Predictions of the radiobiological effectiveness (RBE) play an essential role in treatment planning with heavy charged particles. Amorphous track models ( [1] , [2] , also referred to as track structure models) provide currently the most suitable description of cell survival under i....... Amorphous track modelling of luminescence detector efficiency in proton and carbon beams. 4.Tsuruoka C, Suzuki M, Kanai T, et al. LET and ion species dependence for cell killing in normal human skin fibroblasts. Radiat Res. 2005;163:494-500.......Introduction: Predictions of the radiobiological effectiveness (RBE) play an essential role in treatment planning with heavy charged particles. Amorphous track models ( [1] , [2] , also referred to as track structure models) provide currently the most suitable description of cell survival under ion....... [2] . In addition, a new approach based on microdosimetric distributions is presented and investigated [3] . Material and methods: A suitable software library embrasing the mentioned amorphous track models including numerous submodels with respect to delta-electron range models, radial dose...

  7. Gold nanoparticles for non-invasive cell tracking with CT imaging

    Meir, Rinat; Betzer, Oshra; Barnoy, Eran; Motiei, Menachem; Popovtzer, Rachela

    2018-02-01

    Cell-based therapies use living cells with therapeutic traits to treat various diseases. This is a beneficial alternative for diseases that existing medicine cannot cure efficiently. However, inconsistent results in clinical trials are preventing the advancement and implementation of cell-based therapy. In order to explain such results, there is a need to discover the fate of the transplanted cells. To answer this need, we developed a technique for noninvasive in vivo cell tracking, which uses gold nanoparticles as contrast agents for CT imaging. Herein, we investigate the design principles of this technique for intramuscular transplantation of therapeutic cells. Longitudinal studies were performed, demonstrating the ability to track cells over long periods of time. As few as 500 cells could be detected and a way to quantify the number of cells visualized by CT was demonstrated. This cell-tracking technology has the potential to become an essential tool in pre-clinical studies as well as in clinical trials and advance cell therapy.

  8. The probabilities of one- and multi-track events for modeling radiation-induced cell kill

    Schneider, Uwe; Vasi, Fabiano; Besserer, Juergen [University of Zuerich, Department of Physics, Science Faculty, Zurich (Switzerland); Radiotherapy Hirslanden, Zurich (Switzerland)

    2017-08-15

    In view of the clinical importance of hypofractionated radiotherapy, track models which are based on multi-hit events are currently reinvestigated. These models are often criticized, because it is believed that the probability of multi-track hits is negligible. In this work, the probabilities for one- and multi-track events are determined for different biological targets. The obtained probabilities can be used with nano-dosimetric cluster size distributions to obtain the parameters of track models. We quantitatively determined the probabilities for one- and multi-track events for 100, 500 and 1000 keV electrons, respectively. It is assumed that the single tracks are statistically independent and follow a Poisson distribution. Three different biological targets were investigated: (1) a DNA strand (2 nm scale); (2) two adjacent chromatin fibers (60 nm); and (3) fiber loops (300 nm). It was shown that the probabilities for one- and multi-track events are increasing with energy, size of the sensitive target structure, and dose. For a 2 x 2 x 2 nm{sup 3} target, one-track events are around 10,000 times more frequent than multi-track events. If the size of the sensitive structure is increased to 100-300 nm, the probabilities for one- and multi-track events are of the same order of magnitude. It was shown that target theories can play a role for describing radiation-induced cell death if the targets are of the size of two adjacent chromatin fibers or fiber loops. The obtained probabilities can be used together with the nano-dosimetric cluster size distributions to determine model parameters for target theories. (orig.)

  9. Using Single-Protein Tracking to Study Cell Migration.

    Orré, Thomas; Mehidi, Amine; Massou, Sophie; Rossier, Olivier; Giannone, Grégory

    2018-01-01

    To get a complete understanding of cell migration, it is critical to study its orchestration at the molecular level. Since the recent developments in single-molecule imaging, it is now possible to study molecular phenomena at the single-molecule level inside living cells. In this chapter, we describe how such approaches have been and can be used to decipher molecular mechanisms involved in cell migration.

  10. Particle image velocimetry a practical guide

    Raffel, Marcus; Wereley, Steve T; Kompenhans, Jürgen

    2007-01-01

    The development of Particle Image Velocimetry (PIV), a measurement technique, which allows for capturing velocity information of whole ?ow ?elds in fractions of a second, has begun in the eighties of the last century. In 1998, when this book has been published ?rstly, the PIV technique emerged from laboratories to applications in fundamental and industrial research, in par- lel to the transition from photo-graphicalto video recording techniques. Thus this book, whose objective was and is to serve as a practical guide to the PIV technique, found strong interest within the increasing group of us

  11. Clinically viable magnetic poly(lactide-co-glycolide) (PLGA) particles for MRI-based cell tracking

    Granot, Dorit; Nkansah, Michael K.; Bennewitz, Margaret F.; Tang, Kevin S.; Markakis, Eleni A.; Shapiro, Erik M.

    2013-01-01

    Purpose To design, fabricate, characterize and in vivo assay clinically viable magnetic particles for MRI-based cell tracking. Methods PLGA encapsulated magnetic nano- and microparticles were fabricated. Multiple biologically relevant experiments were performed to assess cell viability, cellular performance and stem cell differentiation. In vivo MRI experiments were performed to separately test cell transplantation and cell migration paradigms, as well as in vivo biodegradation. Results Highly magnetic nano- (~100 nm) and microparticles (~1–2 μm) were fabricated. Magnetic cell labeling in culture occurred rapidly achieving 3–50 pg Fe/cell at 3 hrs for different particles types, and >100 pg Fe/cell after 10 hours, without the requirement of a transfection agent, and with no effect on cell viability. The capability of magnetically labeled mesenchymal or neural stem cells to differentiate down multiple lineages, or for magnetically labeled immune cells to release cytokines following stimulation, was uncompromised. An in vivo biodegradation study revealed that NPs degraded ~80% over the course of 12 weeks. MRI detected as few as 10 magnetically labeled cells, transplanted into the brains of rats. Also, these particles enabled the in vivo monitoring of endogenous neural progenitor cell migration in rat brains over 2 weeks. Conclusion The robust MRI properties and benign safety profile of these particles make them promising candidates for clinical translation for MRI-based cell tracking. PMID:23568825

  12. Tracking and Finding Slow-Proliferating/Quiescent Cancer Stem Cells with Fluorescent Nanodiamonds.

    Lin, Hsin-Hung; Lee, Hsiao-Wen; Lin, Ruey-Jen; Huang, Chih-Wei; Liao, Yi-Chun; Chen, Yit-Tsong; Fang, Jim-Min; Lee, Te-Chang; Yu, Alice L; Chang, Huan-Cheng

    2015-09-09

    Quiescent cancer stem cells (CSCs) have long been considered to be a source of tumor initiation. However, identification and isolation of these cells have been hampered by the fact that commonly used fluorescent markers are not sufficiently stable, both chemically and photophysically, to allow tracking over an extended period of time. Here, it is shown that fluorescent nanodiamonds (FNDs) are well suited for this application. Genotoxicity tests of FNDs with comet and micronucleus assays for human fibroblasts and breast cancer cells indicate that the nanoparticles neither cause DNA damage nor impair cell growth. Using AS-B145-1R breast cancer cells as the model cell line for CSC, it is found that the FND labeling outperforms 5-ethynyl-2'-deoxyuridine (EdU) and carboxyfluorescein diacetate succinimidyl ester (CFSE) in regards to its long-term tracking capability (>20 d). Moreover, through a quantification of their stem cell activity by measuring mammosphere-forming efficiencies (MFEs) and self-renewal rates, the FND-positive cells are identified to have an MFE twice as high as that of the FND-negative cells isolated from the same dissociated mammospheres. Thus, the nanoparticle-based labeling technique provides an effective new tool for tracking and finding slow-proliferating/quiescent CSCs in cancer research. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Krypton tagging velocimetry of an underexpanded jet.

    Parziale, N J; Smith, M S; Marineau, E C

    2015-06-01

    In this work, we present the excitation/emission strategy, experimental setup, and results of an implementation of krypton tagging velocimetry (KTV). KTV is performed as follows: (i) seed a base flow with krypton; (ii) photosynthesize metastable krypton atoms with a frequency-doubled dye laser to form the tagged tracer; (iii) record the translation of the tagged metastable krypton by imaging the laser-induced fluorescence (LIF) that is produced with an additional dye laser. The principle strength of KTV, relative to other tagging velocimetry techniques, is the use of a chemically inert tracer. KTV results are presented for an underexpanded jet of three mixtures of varying Kr/N2 concentration. It is demonstrated that KTV can be used in gas mixtures of relatively low krypton mole fraction (0.5% Kr/99.5% N2), and the KTV data from that experiment are found to be in good agreement with an empirical fit found in the literature. We find that KTV is useful to perform instantaneous velocity measurements with metastable krypton as a chemically inert, dilute, long-lifetime tracer in gas-phase flows.

  14. Laser doppler velocimetry and confined flows

    Ilić Jelena T.

    2017-01-01

    Full Text Available Finding the mode, in which two component laser Doppler velocimetry can be applied to flows confined in cylindrical tubes or vessels, was the aim of this study. We have identified principle issues that influence the propagation of laser beams in laser Doppler velocimetry system, applied to flow confined in cylindrical tube. Among them, the most important are influences of fluid and wall refractive indices, wall thickness and internal radius ratio and beam intersection angle. In analysis of the degrees of these influences, we have applied mathematical model, based on geometrical optics. The separation of measurement volumes, that measure different velocity components, has been recognized as the main drawback. To overcome this, we propose a lens with dual focal length – primary focal length for the measurement of one velocity component and secondary focal length for the measurement of the other velocity component. We present here the procedure for calculating the optimal value of secondary focal length, depending on experimental set-up parameters. The mathematical simulation of the application of the dual focal length lens, for chosen cases presented here, confirmed the accuracy of the proposed procedure.

  15. Cellular track model of biological damage to mammalian cell cultures from galactic cosmic rays

    Cucinotta, Francis A.; Katz, Robert; Wilson, John W.; Townsend, Lawrence W.; Nealy, John E.; Shinn, Judy L.

    1991-01-01

    The assessment of biological damage from the galactic cosmic rays (GCR) is a current interest for exploratory class space missions where the highly ionizing, high-energy, high-charge ions (HZE) particles are the major concern. The relative biological effectiveness (RBE) values determined by ground-based experiments with HZE particles are well described by a parametric track theory of cell inactivation. Using the track model and a deterministic GCR transport code, the biological damage to mammalian cell cultures is considered for 1 year in free space at solar minimum for typical spacecraft shielding. Included are the effects of projectile and target fragmentation. The RBE values for the GCR spectrum which are fluence-dependent in the track model are found to be more severe than the quality factors identified by the International Commission on Radiological Protection publication 26 and seem to obey a simple scaling law with the duration period in free space.

  16. Cellular track model of biological damage to mammalian cell cultures from galactic cosmic rays

    Cucinotta, F.A.; Katz, R.; Wilson, J.W.; Townsend, L.W.; Nealy, J.E.; Shinn, J.L.

    1991-02-01

    The assessment of biological damage from the galactic cosmic rays (GCR) is a current interest for exploratory class space missions where the highly ionizing, high-energy, high-charge ions (HZE) particles are the major concern. The relative biological effectiveness (RBE) values determined by ground-based experiments with HZE particles are well described by a parametric track theory of cell inactivation. Using the track model and a deterministic GCR transport code, the biological damage to mammalian cell cultures is considered for 1 year in free space at solar minimum for typical spacecraft shielding. Included are the effects of projectile and target fragmentation. The RBE values for the GCR spectrum which are fluence-dependent in the track model are found to be more severe than the quality factors identified by the International Commission on Radiological Protection publication 26 and seem to obey a simple scaling law with the duration period in free space

  17. Tracking the Evolution of Non-Small-Cell Lung Cancer

    Jamal-Hanjani, Mariam; Wilson, Gareth A.; McGranahan, Nicholas

    2017-01-01

    Background Among patients with non-small-cell lung cancer (NSCLC), data on intratumor heterogeneity and cancer genome evolution have been limited to small retrospective cohorts. We wanted to prospectively investigate intratumor heterogeneity in relation to clinical outcome and to determine...... as a prognostic predictor. (Funded by Cancer Research UK and others; TRACERx ClinicalTrials.gov number, NCT01888601 .)....

  18. Lightning Jump Algorithm and Relation to Thunderstorm Cell Tracking, GLM Proxy and Other Meteorological Measurements

    Schultz, Christopher J.; Carey, Lawrence D.; Cecil, Daniel J.; Bateman, Monte

    2012-01-01

    The lightning jump algorithm has a robust history in correlating upward trends in lightning to severe and hazardous weather occurrence. The algorithm uses the correlation between the physical principles that govern an updraft's ability to produce microphysical and kinematic conditions conducive for electrification and its role in the development of severe weather conditions. Recent work has demonstrated that the lightning jump algorithm concept holds significant promise in the operational realm, aiding in the identification of thunderstorms that have potential to produce severe or hazardous weather. However, a large amount of work still needs to be completed in spite of these positive results. The total lightning jump algorithm is not a stand-alone concept that can be used independent of other meteorological measurements, parameters, and techniques. For example, the algorithm is highly dependent upon thunderstorm tracking to build lightning histories on convective cells. Current tracking methods show that thunderstorm cell tracking is most reliable and cell histories are most accurate when radar information is incorporated with lightning data. In the absence of radar data, the cell tracking is a bit less reliable but the value added by the lightning information is much greater. For optimal application, the algorithm should be integrated with other measurements that assess storm scale properties (e.g., satellite, radar). Therefore, the recent focus of this research effort has been assessing the lightning jump's relation to thunderstorm tracking, meteorological parameters, and its potential uses in operational meteorology. Furthermore, the algorithm must be tailored for the optically-based GOES-R Geostationary Lightning Mapper (GLM), as what has been observed using Very High Frequency Lightning Mapping Array (VHF LMA) measurements will not exactly translate to what will be observed by GLM due to resolution and other instrument differences. Herein, we present some of

  19. Automatic tracking of red blood cells in micro channels using OpenCV

    Rodrigues, Vânia; Rodrigues, Pedro J.; Pereira, Ana I.; Lima, Rui

    2013-10-01

    The present study aims to developan automatic method able to track red blood cells (RBCs) trajectories flowing through a microchannel using the Open Source Computer Vision (OpenCV). The developed method is based on optical flux calculation assisted by the maximization of the template-matching product. The experimental results show a good functional performance of this method.

  20. Hydroxyl tagging velocimetry in a supersonic flow over a cavity

    Pitz, Robert W.; Lahr, Michael D.; Douglas, Zachary W.; Wehrmeyer, Joseph A.; Hu Shengteng; Carter, Campbell D.; Hsu, Kuang-Yu; Lum, Chee; Koochesfahani, Manoochehr M.

    2005-01-01

    Hydroxyl tagging velocimetry (HTV) measurements of velocity were made in a Mach 2 (M 2) flow with a wall cavity. In the HTV method, ArF excimer laser (193 nm) beams pass through a humid gas and dissociate H2O into H + OH to form a tagging grid of OH molecules. In this study, a 7x7 grid of hydroxyl (OH) molecules is tracked by planar laser-induced fluorescence. The grid motion over a fixed time delay yields about 50 velocity vectors of the two-dimensional flow in the plane of the laser sheets. Velocity precision is limited by the error in finding the crossing location of the OH lines written by the excimer tag laser. With a signal-to-noise ratio of about 10 for the OH lines, the determination of the crossing location is expected to be accurate within ±0.1 pixels. Velocity precision within the freestream, where the turbulence is low, is consistent with this error. Instantaneous, single-shot measurements of two-dimensional flow patterns were made in the nonreacting M 2 flow with a wall cavity under low- and high-pressure conditions. The single-shot profiles were analyzed to yield mean and rms velocity profiles in the M 2 nonreacting flow

  1. Cancer cell imaging by stable wet near-field scanning optical microscope with resonance tracking method

    Park, Kyoung-Duck; Park, Doo-Jae; Jeong, Mun-Seok; Choi, Geun-Chang; Lee, Seung-Gol; Byeon, Clare-Chisu; Choi, Soo-Bong

    2014-01-01

    We report on a successful topographical and optical imaging of various cancer cells in liquid and in air by using a stable wet near-field scanning optical microscope that utilizes a resonance tracking method. We observed a clear dehydration which gives rise to a decrease in the cell volume down to 51%. In addition, a micro-ball lens effect due to the round-shaped young cancer cells was observed from near-field imaging, where the refractive index of young cancer cells was deduced.

  2. Cancer cell imaging by stable wet near-field scanning optical microscope with resonance tracking method

    Park, Kyoung-Duck [Sungkyunkwan University, Suwon (Korea, Republic of); Inha University, Incheon (Korea, Republic of); Park, Doo-Jae; Jeong, Mun-Seok [Sungkyunkwan University, Suwon (Korea, Republic of); Choi, Geun-Chang [Seoul National University, Seoul (Korea, Republic of); Lee, Seung-Gol [Inha University, Incheon (Korea, Republic of); Byeon, Clare-Chisu [Kyungpook National University, Daegu (Korea, Republic of); Choi, Soo-Bong [Incheon National University, Incheon (Korea, Republic of)

    2014-05-15

    We report on a successful topographical and optical imaging of various cancer cells in liquid and in air by using a stable wet near-field scanning optical microscope that utilizes a resonance tracking method. We observed a clear dehydration which gives rise to a decrease in the cell volume down to 51%. In addition, a micro-ball lens effect due to the round-shaped young cancer cells was observed from near-field imaging, where the refractive index of young cancer cells was deduced.

  3. Attachment and spreadout study of 3T3 cells onto PP track etched films

    Smolko, Eduardo; Mazzei, Ruben; Tadey, Daniel; Lombardo, Daniel

    2001-01-01

    Polymer surface modifications are obtained by the application of radiation treatments and other physico-chemical methods: fission fragment (ff) irradiation and etching. The biocompatibility of the surface is then observed by cell seeding and cell adhesion experiments. Approaches to improvement of the cell adhesion are obtained by different methods: for example, in PS, cell adhesion is improved after ion implantation; in PMMA, after bombarding the polymer, the surface is reconditioned with surfactants and proteins and in PVDF, cell adhesion is assayed on nuclear tracks membranes. In this work, we obtained important cell adhesion improvements in PP films by irradiation with swift heavy ions and subsequent etching of the nuclear tracks. We use BOPP (isotactic -25 μm thickness). Irrradiations were performed with a Cf-252 californium ff source. The source has a heavy ff and a light one, with 160-200 MeV energy divided among them corresponding to ff energies between 1 and 2 MeV/amu. A chemical etching procedure consisting of a solution of sulphuric acid and chromium three oxide at 85 deg. C was used. The 3T3 NIH fibroblast cell line was used for the cell adhesion experiment. Here we report for the first time, the results of a series of experiments by varying the ff fluence and the etching time showing that attachment and spreadout of cells are very much improved in this cell line according to the number of pores and the pore size

  4. Guard cell zeaxanthin tracks photosynthetically active radiation and stomatal apertures in Vicia faba leaves

    Srivastava, A.; Zeiger, E.

    1995-01-01

    Zeaxanthin, antheraxanthin and violaxanthin concentrations in guard cells from sonicated abaxial epidermal peels of Vicia faba were measured from dawn to dusk, and compared with concentrations in mesophyll tissue of the same leaves. Measured changes in guard cell zeaxanthin and violaxanthin concentrations indicate that guard cells operate the xanthophyll cycle throughout the day. Mesophyll tissue had no detectable zeaxanthin at dawn, whereas guard cells had 30–50 mmol mol −1 chlorophyll a+b. On a chlorophyll basis, maximal zeaxanthin levels were 3–4 fold higher in guard cells than in mesophyll cells. Zeaxanthin concentrations tracked levels of photosynthetically active radiation (PAR) in both mesophyll and guard cells. In the mesophyll, most of the zeaxanthin changes occurred in mid-morning and mid-afternoon. In guard cells, zeaxanthin concentrations changed nearly linearly with PAR in the early morning and late afternoon, and closely tracked PAR levels throughout the day. Guard cell zeaxanthin concentrations were also closely correlated with stomatal apertures. The close relationship between zeaxanthin concentrations and PAR levels in guard cells indicates that zeaxanthin is well suited to function as a molecular photosensor in stomatal movements. (author)

  5. Convolutional Deep Belief Networks for Single-Cell/Object Tracking in Computational Biology and Computer Vision.

    Zhong, Bineng; Pan, Shengnan; Zhang, Hongbo; Wang, Tian; Du, Jixiang; Chen, Duansheng; Cao, Liujuan

    2016-01-01

    In this paper, we propose deep architecture to dynamically learn the most discriminative features from data for both single-cell and object tracking in computational biology and computer vision. Firstly, the discriminative features are automatically learned via a convolutional deep belief network (CDBN). Secondly, we design a simple yet effective method to transfer features learned from CDBNs on the source tasks for generic purpose to the object tracking tasks using only limited amount of training data. Finally, to alleviate the tracker drifting problem caused by model updating, we jointly consider three different types of positive samples. Extensive experiments validate the robustness and effectiveness of the proposed method.

  6. Cell Membrane Tracking in Living Brain Tissue Using Differential Interference Contrast Microscopy.

    Lee, John; Kolb, Ilya; Forest, Craig R; Rozell, Christopher J

    2018-04-01

    Differential interference contrast (DIC) microscopy is widely used for observing unstained biological samples that are otherwise optically transparent. Combining this optical technique with machine vision could enable the automation of many life science experiments; however, identifying relevant features under DIC is challenging. In particular, precise tracking of cell boundaries in a thick ( ) slice of tissue has not previously been accomplished. We present a novel deconvolution algorithm that achieves the state-of-the-art performance at identifying and tracking these membrane locations. Our proposed algorithm is formulated as a regularized least squares optimization that incorporates a filtering mechanism to handle organic tissue interference and a robust edge-sparsity regularizer that integrates dynamic edge tracking capabilities. As a secondary contribution, this paper also describes new community infrastructure in the form of a MATLAB toolbox for accurately simulating DIC microscopy images of in vitro brain slices. Building on existing DIC optics modeling, our simulation framework additionally contributes an accurate representation of interference from organic tissue, neuronal cell-shapes, and tissue motion due to the action of the pipette. This simulator allows us to better understand the image statistics (to improve algorithms), as well as quantitatively test cell segmentation and tracking algorithms in scenarios, where ground truth data is fully known.

  7. Objective speckle velocimetry for autonomous vehicle odometry.

    Francis, D; Charrett, T O H; Waugh, L; Tatam, R P

    2012-06-01

    Speckle velocimetry is investigated as a means of determining odometry data with potential for application on autonomous robotic vehicles. The technique described here relies on the integration of translation measurements made by normalized cross-correlation of speckle patterns to determine the change in position over time. The use of objective (non-imaged) speckle offers a number of advantages over subjective (imaged) speckle, such as a reduction in the number of optical components, reduced modulation of speckles at the edges of the image, and improved light efficiency. The influence of the source/detector configuration on the speckle translation to vehicle translation scaling factor for objective speckle is investigated using a computer model and verified experimentally. Experimental measurements are presented at velocities up to 80  mm s(-1) which show accuracy better than 0.4%.

  8. An Evaluation of an Automatic Cell Detection and Tracking Algorithm.

    1982-11-03

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  9. Cell tracking using iron oxide fails to distinguish dead from living transplanted cells in the infarcted heart.

    Winter, E M; Hogers, B; van der Graaf, L M; Gittenberger-de Groot, A C; Poelmann, R E; van der Weerd, L

    2010-03-01

    Recently, debate has arisen about the usefulness of cell tracking using iron oxide-labeled cells. Two important issues in determining the usefulness of cell tracking with MRI are generally overlooked; first, the effect of graft rejection in immunocompetent models, and second, the necessity for careful histological confirmation of the fate of the labeled cells in the presence of iron oxide. Therefore, both iron oxide-labeled living as well as dead epicardium-derived cells (EPDCs) were investigated in ischemic myocardium of immunodeficient non-obese diabetic (NOD)/acid: non-obese diabetic severe combined immunodeficient (NOD/scid) mice with 9.4T MRI until 6 weeks after surgery, at which time immunohistochemical analysis was performed. In both groups, voids on MRI scans were observed that did not change in number, size, or localization over time. Based on MRI, no distinction could be made between living and dead injected cells. Prussian blue staining confirmed that the hypointense spots on MRI corresponded to iron-loaded cells. However, in the dead-EPDC recipients, all iron-positive cells appeared to be macrophages, while the living-EPDC recipients also contained engrafted iron-loaded EPDCs. Iron labeling is inadequate for determining the fate of transplanted cells in the immunodeficient host, since dead cells produce an MRI signal indistinguishable from incorporated living cells. (c) 2010 Wiley-Liss, Inc.

  10. Particle image velocimetry - Principles and first results

    Laporta, A.; Marechal, J.P.

    1997-01-01

    Particle Image Velocimetry (PIV) is a measurement technique elaborated towards the end of the 1970's, but which has developed considerably in recent years. The general principle of PIV is very simple and enables access to instantaneous velocity fields. It consists in recording images of tracer-particles injected into the flow and determining the distance covered by these particles. Since we know the time lapse between successive images of the same particle, we can derive the local fluid velocity. Among the many existing image acquisition and processing methods, the image inter-correlation analysis techniques, used with a pulsed laser source, is the most effective. Since we know the influence of different parameters (number of particles, beam power, time lapse between two successive images, size of query zones, etc.) on the quality of the final result, we can optimize practical adjustment of the PIV measurement scheme. The PIV was tested on the LAVITA hydraulic mockup (simulating the operation of a tangential fan). First results are, all in all, highly satisfactory. These have enabled the rapid drafting of instantaneous mean velocity field maps (20 images acquired in less than 90 seconds, with a post-processing time of about 10 minutes). Observation of the instantaneous fields has evidenced the presence of low frequency non-stationary phenomena which are not revealed by Laser Doppler Velocimetry (LDV). Quantitative comparison between LDV and PIV, concerning average fields, showed close results, with, however, local divergences which could be relatively marked. It must nevertheless be noted that the PIV measurements performed on LAVITA have not been optimized with a view to obtaining a consistently good accuracy level. Priority in the present case was given to the scope of the field explored, with a view to observing the large non-stationary structures within a flow. The PIV measurement technique is thus operational for prompt flow characterization. However

  11. Innovative molecular-based fluorescent nanoparticles for multicolor single particle tracking in cells

    Daniel, Jonathan; Blanchard-Desce, Mireille; Godin, Antoine G; Palayret, Matthieu; Lounis, Brahim; Cognet, Laurent

    2016-01-01

    Based on an original molecular-based design, we present bright and photostable fluorescent organic nanoparticles (FONs) showing excellent colloidal stability in various aqueous environments. Complementary near-infrared emitting and green emitting FONs were prepared using a simple, fast and robust protocol. Both types of FONs could be simultaneously imaged at the single-particle level in solution as well as in biological environments using a monochromatic excitation and a dual-color fluorescence microscope. No evidence of acute cytotoxicity was found upon incubation of live cells with mixed solutions of FONs, and both types of nanoparticles were found internalized in the cells where their motion could be simultaneously tracked at video-rate up to minutes. These fluorescent organic nanoparticles open a novel non-toxic alternative to existing nanoparticles for imaging biological structures, compatible with live-cell experiments and specially fitted for multicolor single particle tracking. (paper)

  12. Effects of track structure and cell inactivation on the calculation of heavy ion mutation rates in mammalian cells

    Cucinotta, F. A.; Wilson, J. W.; Shavers, M. R.; Katz, R.

    1996-01-01

    It has long been suggested that inactivation severely effects the probability of mutation by heavy ions in mammalian cells. Heavy ions have observed cross sections of inactivation that approach and sometimes exceed the geometric size of the cell nucleus in mammalian cells. In the track structure model of Katz the inactivation cross section is found by summing an inactivation probability over all impact parameters from the ion to the sensitive sites within the cell nucleus. The inactivation probability is evaluated using the dose-response of the system to gamma-rays and the radial dose of the ions and may be equal to unity at small impact parameters for some ions. We show how the effects of inactivation may be taken into account in the evaluation of the mutation cross sections from heavy ions in the track structure model through correlation of sites for gene mutation and cell inactivation. The model is fit to available data for HPRT mutations in Chinese hamster cells and good agreement is found. The resulting calculations qualitatively show that mutation cross sections for heavy ions display minima at velocities where inactivation cross sections display maxima. Also, calculations show the high probability of mutation by relativistic heavy ions due to the radial extension of ions track from delta-rays in agreement with the microlesion concept. The effects of inactivation on mutations rates make it very unlikely that a single parameter such as LET or Z*2/beta(2) can be used to specify radiation quality for heavy ion bombardment.

  13. Low-cost motility tracking system (LOCOMOTIS for time-lapse microscopy applications and cell visualisation.

    Adam E Lynch

    Full Text Available Direct visualisation of cells for the purpose of studying their motility has typically required expensive microscopy equipment. However, recent advances in digital sensors mean that it is now possible to image cells for a fraction of the price of a standard microscope. Along with low-cost imaging there has also been a large increase in the availability of high quality, open-source analysis programs. In this study we describe the development and performance of an expandable cell motility system employing inexpensive, commercially available digital USB microscopes to image various cell types using time-lapse and perform tracking assays in proof-of-concept experiments. With this system we were able to measure and record three separate assays simultaneously on one personal computer using identical microscopes, and obtained tracking results comparable in quality to those from other studies that used standard, more expensive, equipment. The microscopes used in our system were capable of a maximum magnification of 413.6×. Although resolution was lower than that of a standard inverted microscope we found this difference to be indistinguishable at the magnification chosen for cell tracking experiments (206.8×. In preliminary cell culture experiments using our system, velocities (mean µm/min ± SE of 0.81 ± 0.01 (Biomphalaria glabrata hemocytes on uncoated plates, 1.17 ± 0.004 (MDA-MB-231 breast cancer cells, 1.24 ± 0.006 (SC5 mouse Sertoli cells and 2.21 ± 0.01 (B. glabrata hemocytes on Poly-L-Lysine coated plates, were measured and are consistent with previous reports. We believe that this system, coupled with open-source analysis software, demonstrates that higher throughput time-lapse imaging of cells for the purpose of studying motility can be an affordable option for all researchers.

  14. Low-cost motility tracking system (LOCOMOTIS) for time-lapse microscopy applications and cell visualisation.

    Lynch, Adam E; Triajianto, Junian; Routledge, Edwin

    2014-01-01

    Direct visualisation of cells for the purpose of studying their motility has typically required expensive microscopy equipment. However, recent advances in digital sensors mean that it is now possible to image cells for a fraction of the price of a standard microscope. Along with low-cost imaging there has also been a large increase in the availability of high quality, open-source analysis programs. In this study we describe the development and performance of an expandable cell motility system employing inexpensive, commercially available digital USB microscopes to image various cell types using time-lapse and perform tracking assays in proof-of-concept experiments. With this system we were able to measure and record three separate assays simultaneously on one personal computer using identical microscopes, and obtained tracking results comparable in quality to those from other studies that used standard, more expensive, equipment. The microscopes used in our system were capable of a maximum magnification of 413.6×. Although resolution was lower than that of a standard inverted microscope we found this difference to be indistinguishable at the magnification chosen for cell tracking experiments (206.8×). In preliminary cell culture experiments using our system, velocities (mean µm/min ± SE) of 0.81 ± 0.01 (Biomphalaria glabrata hemocytes on uncoated plates), 1.17 ± 0.004 (MDA-MB-231 breast cancer cells), 1.24 ± 0.006 (SC5 mouse Sertoli cells) and 2.21 ± 0.01 (B. glabrata hemocytes on Poly-L-Lysine coated plates), were measured and are consistent with previous reports. We believe that this system, coupled with open-source analysis software, demonstrates that higher throughput time-lapse imaging of cells for the purpose of studying motility can be an affordable option for all researchers.

  15. Low-Cost Motility Tracking System (LOCOMOTIS) for Time-Lapse Microscopy Applications and Cell Visualisation

    Lynch, Adam E.; Triajianto, Junian; Routledge, Edwin

    2014-01-01

    Direct visualisation of cells for the purpose of studying their motility has typically required expensive microscopy equipment. However, recent advances in digital sensors mean that it is now possible to image cells for a fraction of the price of a standard microscope. Along with low-cost imaging there has also been a large increase in the availability of high quality, open-source analysis programs. In this study we describe the development and performance of an expandable cell motility system employing inexpensive, commercially available digital USB microscopes to image various cell types using time-lapse and perform tracking assays in proof-of-concept experiments. With this system we were able to measure and record three separate assays simultaneously on one personal computer using identical microscopes, and obtained tracking results comparable in quality to those from other studies that used standard, more expensive, equipment. The microscopes used in our system were capable of a maximum magnification of 413.6×. Although resolution was lower than that of a standard inverted microscope we found this difference to be indistinguishable at the magnification chosen for cell tracking experiments (206.8×). In preliminary cell culture experiments using our system, velocities (mean µm/min ± SE) of 0.81±0.01 (Biomphalaria glabrata hemocytes on uncoated plates), 1.17±0.004 (MDA-MB-231 breast cancer cells), 1.24±0.006 (SC5 mouse Sertoli cells) and 2.21±0.01 (B. glabrata hemocytes on Poly-L-Lysine coated plates), were measured and are consistent with previous reports. We believe that this system, coupled with open-source analysis software, demonstrates that higher throughput time-lapse imaging of cells for the purpose of studying motility can be an affordable option for all researchers. PMID:25121722

  16. Contribution of macrophages in the contrast loss in iron oxide-based MRI cancer cell tracking studies

    Danhier, Pierre; Deumer, Gladys; Joudiou, Nicolas; Bouzin, Caroline; Levêque, Philippe; Haufroid, Vincent; Jordan, Bénédicte F.; Feron, Olivier; Sonveaux, Pierre; Gallez, Bernard

    2017-01-01

    Magnetic resonance imaging (MRI) cell tracking of cancer cells labeled with superparamagnetic iron oxides (SPIO) allows visualizing metastatic cells in preclinical models. However, previous works showed that the signal void induced by SPIO on T2(*)-weighted images decreased over time. Here, we aim at characterizing the fate of iron oxide nanoparticles used in cell tracking studies and the role of macrophages in SPIO metabolism. In vivo MRI cell tracking of SPIO positive 4T1 breast cancer cells revealed a quick loss of T2* contrast after injection. We next took advantage of electron paramagnetic resonance (EPR) spectroscopy and inductively coupled plasma mass spectroscopy (ICP-MS) for characterizing the evolution of superparamagnetic and non-superparamagnetic iron pools in 4T1 breast cancer cells and J774 macrophages after SPIO labeling. These in vitro experiments and histology studies performed on 4T1 tumors highlighted the quick degradation of iron oxides by macrophages in SPIO-based cell tracking experiments. In conclusion, the release of SPIO by dying cancer cells and the subsequent uptake of iron oxides by tumor macrophages are limiting factors in MRI cell tracking experiments that plead for the use of (MR) reporter-gene based imaging methods for the long-term tracking of metastatic cells. PMID:28467814

  17. Automatic tracking of cells for video microscopy in patch clamp experiments.

    Peixoto, Helton M; Munguba, Hermany; Cruz, Rossana M S; Guerreiro, Ana M G; Leao, Richardson N

    2014-06-20

    Visualisation of neurons labeled with fluorescent proteins or compounds generally require exposure to intense light for a relatively long period of time, often leading to bleaching of the fluorescent probe and photodamage of the tissue. Here we created a technique to drastically shorten light exposure and improve the targeting of fluorescent labeled cells that is specially useful for patch-clamp recordings. We applied image tracking and mask overlay to reduce the time of fluorescence exposure and minimise mistakes when identifying neurons. Neurons are first identified according to visual criteria (e.g. fluorescence protein expression, shape, viability etc.) and a transmission microscopy image Differential Interference Contrast (DIC) or Dodt contrast containing the cell used as a reference for the tracking algorithm. A fluorescence image can also be acquired later to be used as a mask (that can be overlaid on the target during live transmission video). As patch-clamp experiments require translating the microscope stage, we used pattern matching to track reference neurons in order to move the fluorescence mask to match the new position of the objective in relation to the sample. For the image processing we used the Open Source Computer Vision (OpenCV) library, including the Speeded-Up Robust Features (SURF) for tracking cells. The dataset of images (n = 720) was analyzed under normal conditions of acquisition and with influence of noise (defocusing and brightness). We validated the method in dissociated neuronal cultures and fresh brain slices expressing Enhanced Yellow Fluorescent Protein (eYFP) or Tandem Dimer Tomato (tdTomato) proteins, which considerably decreased the exposure to fluorescence excitation, thereby minimising photodamage. We also show that the neuron tracking can be used in differential interference contrast or Dodt contrast microscopy. The techniques of digital image processing used in this work are an important addition to the set of microscopy

  18. Three-dimensional single-particle tracking in live cells: news from the third dimension

    Dupont, A; Wehnekamp, F; Katayama, Y; Lamb, D C; Gorelashvili, M; Schüller, V; Arcizet, D; Heinrich, D

    2013-01-01

    Single-particle tracking (SPT) is of growing importance in the biophysical community. It is used to investigate processes such as drug and gene delivery, viral uptake, intracellular trafficking or membrane-bound protein mobility. Traditionally, SPT is performed in two dimensions (2D) because of its technical simplicity. However, life occurs in three dimensions (3D) and many methods have been recently developed to track particles in 3D. Now, is the third dimension worth the effort? Here we investigate the differences between the 2D and 3D analyses of intracellular transport with the 3D development of a time-resolved mean square displacement (MSD) analysis introduced previously. The 3D trajectories, and the 2D projections, of fluorescent nanoparticles were obtained with an orbital tracking microscope in two different cell types: in Dictyostelium discoideum ameba and in adherent, more flattened HuH-7 human cells. As expected from the different 3D organization of both cells’ cytoskeletons, a third of the active transport was lost upon projection in the ameba whereas the identification of the active phases was barely affected in the HuH-7 cells. In both cell types, we found intracellular diffusion to be anisotropic and the diffusion coefficient values derived from the 2D analysis were therefore biased. (paper)

  19. Paramagnetic particles carried by cell-penetrating peptide tracking of bone marrow mesenchymal stem cells, a research in vitro

    Liu Min; Guo Youmin; Wu Qifei; Yang Junle; Wang Peng; Wang Sicen; Guo Xiaojuan; Qiang Yongqian; Duan Xiaoyi

    2006-01-01

    The ability to track the distribution and differentiation of stem cells by high-resolution imaging techniques would have significant clinical and research implications. In this study, a model cell-penetrating peptide was used to carry gadolinium particles for magnetic resonance imaging of the mesenchymal stem cells. The mesenchymal stem cells were isolated from rat bone marrow by Percoll and identified by osteogenic differentiation in vitro. The cell-penetrating peptides labeled with fluorescein-5-isothiocyanate and gadolinium were synthesized by a solid-phase peptide synthesis method and the relaxivity of cell-penetrating peptide-gadolinium paramagnetic conjugate on 400 MHz nuclear magnetic resonance was 5.7311 ± 0.0122 mmol -1 s -1 , higher than that of diethylenetriamine pentaacetic acid gadolinium (p < 0.05). Fluorescein imaging confirmed that this new peptide could internalize into the cytoplasm and nucleus. Gadolinium was efficiently internalized into mesenchymal stem cells by the peptide in a time- or concentration-dependent fashion, resulting in intercellular T1 relaxation enhancement, which was obviously detected by 1.5 T magnetic resonance imaging. Cytotoxicity assay and flow cytometric analysis showed the intercellular contrast medium incorporation did not affect cell viability and membrane potential gradient. The research in vitro suggests that the newly constructed peptides could be a vector for tracking mesenchymal stem cells

  20. Long-term tracking of budding yeast cells in brightfield microscopy: CellStar and the Evaluation Platform.

    Versari, Cristian; Stoma, Szymon; Batmanov, Kirill; Llamosi, Artémis; Mroz, Filip; Kaczmarek, Adam; Deyell, Matt; Lhoussaine, Cédric; Hersen, Pascal; Batt, Gregory

    2017-02-01

    With the continuous expansion of single cell biology, the observation of the behaviour of individual cells over extended durations and with high accuracy has become a problem of central importance. Surprisingly, even for yeast cells that have relatively regular shapes, no solution has been proposed that reaches the high quality required for long-term experiments for segmentation and tracking (S&T) based on brightfield images. Here, we present CellStar , a tool chain designed to achieve good performance in long-term experiments. The key features are the use of a new variant of parametrized active rays for segmentation, a neighbourhood-preserving criterion for tracking, and the use of an iterative approach that incrementally improves S&T quality. A graphical user interface enables manual corrections of S&T errors and their use for the automated correction of other, related errors and for parameter learning. We created a benchmark dataset with manually analysed images and compared CellStar with six other tools, showing its high performance, notably in long-term tracking. As a community effort, we set up a website, the Yeast Image Toolkit, with the benchmark and the Evaluation Platform to gather this and additional information provided by others. © 2017 The Authors.

  1. Photophoretic velocimetry for the characterization of aerosols.

    Haisch, Christoph; Kykal, Carsten; Niessner, Reinhard

    2008-03-01

    Aerosols are particles in a size range from some nanometers to some micrometers suspended in air or other gases. Their relevance varies as wide as their origin and composition. In the earth's atmosphere they influence the global radiation balance and human health. Artificially produced aerosols are applied, e.g., for drug administration, as paint and print pigments, or in rubber tire production. In all these fields, an exact characterization of single particles as well as of the particle ensemble is essential. Beyond characterization, continuous separation is often required. State-of-the-art separation techniques are based on electrical, thermal, or flow fields. In this work we present an approach to apply light in the form of photophoretic (PP) forces for characterization and separation of aerosol particles according to their optical properties. Such separation technique would allow, e.g., the separation of organic from inorganic particles of the same aerodynamic size. We present a system which automatically records velocities induced by PP forces and does a statistical evaluation in order to characterize the particle ensemble properties. The experimental system essentially consists of a flow cell with rectangular cross section (1 cm(2), length 7 cm), where the aerosol stream is pumped through in the vertical direction at ambient pressure. In the cell, a laser beam is directed orthogonally to the particle flow direction, which results in a lateral displacement of the particles. In an alternative configuration, the beam is directed in the opposite direction to the aerosol flow; hence, the particles are slowed down by the PP force. In any case, the photophoretically induced variations of speed and position are visualized by a second laser illumination and a camera system, feeding a mathematical particle tracking algorithm. The light source inducing the PP force is a diode laser (lambda = 806 nm, P = 0.5 W).

  2. Tracking the engraftment and regenerative capabilities of transplanted lung stem cells using fluorescent nanodiamonds.

    Wu, Tsai-Jung; Tzeng, Yan-Kai; Chang, Wei-Wei; Cheng, Chi-An; Kuo, Yung; Chien, Chin-Hsiang; Chang, Huan-Cheng; Yu, John

    2013-09-01

    Lung stem/progenitor cells are potentially useful for regenerative therapy, for example in repairing damaged or lost lung tissue in patients. Several optical imaging methods and probes have been used to track how stem cells incorporate and regenerate themselves in vivo over time. However, these approaches are limited by photobleaching, toxicity and interference from background tissue autofluorescence. Here we show that fluorescent nanodiamonds, in combination with fluorescence-activated cell sorting, fluorescence lifetime imaging microscopy and immunostaining, can identify transplanted CD45(-)CD54(+)CD157(+) lung stem/progenitor cells in vivo, and track their engraftment and regenerative capabilities with single-cell resolution. Fluorescent nanodiamond labelling did not eliminate the cells' properties of self-renewal and differentiation into type I and type II pneumocytes. Time-gated fluorescence imaging of tissue sections of naphthalene-injured mice indicates that the fluorescent nanodiamond-labelled lung stem/progenitor cells preferentially reside at terminal bronchioles of the lungs for 7 days after intravenous transplantation.

  3. Single-cell tracking reveals antibiotic-induced changes in mycobacterial energy metabolism.

    Maglica, Željka; Özdemir, Emre; McKinney, John D

    2015-02-17

    ATP is a key molecule of cell physiology, but despite its importance, there are currently no methods for monitoring single-cell ATP fluctuations in live bacteria. This is a major obstacle in studies of bacterial energy metabolism, because there is a growing awareness that bacteria respond to stressors such as antibiotics in a highly individualistic manner. Here, we present a method for long-term single-cell tracking of ATP levels in Mycobacterium smegmatis based on a combination of microfluidics, time-lapse microscopy, and Förster resonance energy transfer (FRET)-based ATP biosensors. Upon treating cells with antibiotics, we observed that individual cells undergo an abrupt and irreversible switch from high to low intracellular ATP levels. The kinetics and extent of ATP switching clearly discriminate between an inhibitor of ATP synthesis and other classes of antibiotics. Cells that resume growth after 24 h of antibiotic treatment maintain high ATP levels throughout the exposure period. In contrast, antibiotic-treated cells that switch from ATP-high to ATP-low states never resume growth after antibiotic washout. Surprisingly, only a subset of these nongrowing ATP-low cells stains with propidium iodide (PI), a widely used live/dead cell marker. These experiments also reveal a cryptic subset of cells that do not resume growth after antibiotic washout despite remaining ATP high and PI negative. We conclude that ATP tracking is a more dynamic, sensitive, reliable, and discriminating marker of cell viability than staining with PI. This method could be used in studies to evaluate antimicrobial effectiveness and mechanism of action, as well as for high-throughput screening. New antimicrobials are urgently needed to stem the rising tide of antibiotic-resistant bacteria. All antibiotics are expected to affect bacterial energy metabolism, directly or indirectly, yet tools to assess the impact of antibiotics on the ATP content of individual bacterial cells are lacking. The

  4. Quantum dot labeling and tracking of cultured limbal epithelial cell transplants in-vitro

    Genicio, Nuria; Paramo, Juan Gallo; Shortt, Alex J.

    2015-01-01

    PURPOSE Cultured human limbal epithelial cells (HLEC) have shown promise in the treatment of limbal stem cell deficiency but little is known about their survival, behaviour and long-term fate post transplantation. The aim of this research was to evaluate, in-vitro, quantum dot (QDot) technology as a tool for tracking transplanted HLEC. METHODS In-vitro cultured HLEC were labeled with Qdot nanocrystals. Toxicity was assessed using live-dead assays. The effect on HLEC function was assessed using colony forming efficiency assays and expression of CK3, P63alpha and ABCG2. Sheets of cultured HLEC labeled with Qdot nanocrystals were transplanted onto decellularised human corneo-scleral rims in an organ culture model and observed to investigate the behaviour of transplanted cells. RESULTS Qdot labeling had no detrimental effect on HLEC viability or function in-vitro. Proliferation resulted in a gradual reduction in Qdot signal but sufficient signal was present to allow tracking of cells through multiple generations. Cells labeled with Qdots could be reliably detected and observed using confocal microscopy for at least 2 weeks post transplantation in our organ culture model. In addition it was possible to label and observe epithelial cells in intact human corneas using the Rostock corneal module adapted for use with the Heidelberg HRA. CONCLUSIONS This work demonstrates that Qdots combined with existing clinical equipment could be used to track HLEC for up to 2 weeks post transplantation, however, our model does not permit the assessment of cell labeling beyond 2 weeks. Further characterisation in in-vivo models are required. PMID:26024089

  5. Tracking stem cells in tissue-engineered organs using magnetic nanoparticles

    Hachani, R.; Lowdell, M.; Birchall, M.; Thanh, N. T.

    2013-01-01

    The use of human stem cells (SCs) in tissue engineering holds promise in revolutionising the treatment of numerous diseases. There is a pressing need to comprehend the distribution, movement and role of SCs once implanted onto scaffolds. Nanotechnology has provided a platform to investigate this through the development of inorganic magnetic nanoparticles (MNPs). MNPs can be used to label and track SCs by magnetic resonance imaging (MRI) since this clinically available imaging modality has hig...

  6. Fluorescent nanodiamonds enable quantitative tracking of human mesenchymal stem cells in miniature pigs

    Su, Long-Jyun; Wu, Meng-Shiue; Hui, Yuen Yung; Chang, Be-Ming; Pan, Lei; Hsu, Pei-Chen; Chen, Yit-Tsong; Ho, Hong-Nerng; Huang, Yen-Hua; Ling, Thai-Yen; Hsu, Hsao-Hsun; Chang, Huan-Cheng

    2017-03-01

    Cell therapy is a promising strategy for the treatment of human diseases. While the first use of cells for therapeutic purposes can be traced to the 19th century, there has been a lack of general and reliable methods to study the biodistribution and associated pharmacokinetics of transplanted cells in various animal models for preclinical evaluation. Here, we present a new platform using albumin-conjugated fluorescent nanodiamonds (FNDs) as biocompatible and photostable labels for quantitative tracking of human placenta choriodecidual membrane-derived mesenchymal stem cells (pcMSCs) in miniature pigs by magnetic modulation. With this background-free detection technique and time-gated fluorescence imaging, we have been able to precisely determine the numbers as well as positions of the transplanted FND-labeled pcMSCs in organs and tissues of the miniature pigs after intravenous administration. The method is applicable to single-cell imaging and quantitative tracking of human stem/progenitor cells in rodents and other animal models as well.

  7. Helioseismology in a bottle: modal acoustic velocimetry

    Triana, Santiago Andrés; Zimmerman, Daniel S; Lathrop, Daniel P; Nataf, Henri-Claude; Thorette, Aurélien; Lekic, Vedran

    2014-01-01

    Measurement of the differential rotation of the Sun's interior is one of the great achievements of helioseismology, providing important constraints for stellar physics. The technique relies on observing and analyzing rotationally-induced splittings of p-modes in the star. Here, we demonstrate the first use of the technique in a laboratory setting. We apply it in a spherical cavity with a spinning central core (spherical-Couette flow) to determine the mean azimuthal velocity of the air filling the cavity. We excite a number of acoustic resonances (analogous to p-modes in the Sun) using a speaker and record the response with an array of small microphones on the outer sphere. Many observed acoustic modes show rotationally-induced splittings, which allow us to perform an inversion to determine the air's azimuthal velocity as a function of both radius and latitude. We validate the method by comparing the velocity field obtained through inversion against the velocity profile measured with a calibrated hot film anemometer. This modal acoustic velocimetry technique has great potential for laboratory setups involving rotating fluids in axisymmetric cavities. It will be useful especially in liquid metals where direct optical methods are unsuitable and ultrasonic techniques very challenging at best. (paper)

  8. FLEET Velocimetry Measurements on a Transonic Airfoil

    Burns, Ross A.; Danehy, Paul M.

    2017-01-01

    Femtosecond laser electronic excitation tagging (FLEET) velocimetry was used to study the flowfield around a symmetric, transonic airfoil in the NASA Langley 0.3-m TCT facility. A nominal Mach number of 0.85 was investigated with a total pressure of 125 kPa and total temperature of 280 K. Two-components of velocity were measured along vertical profiles at different locations above, below, and aft of the airfoil at angles of attack of 0 deg, 3.5 deg, and 7deg. Measurements were assessed for their accuracy, precision, dynamic range, spatial resolution, and overall measurement uncertainty in the context of the applied flowfield. Measurement precisions as low as 1 m/s were observed, while overall uncertainties ranged from 4 to 5 percent. Velocity profiles within the wake showed sufficient accuracy, precision, and sensitivity to resolve both the mean and fluctuating velocities and general flow physics such as shear layer growth. Evidence of flow separation is found at high angles of attack.

  9. In vivo stem cell tracking with imageable nanoparticles that bind bioorthogonal chemical receptors on the stem cell surface.

    Lee, Sangmin; Yoon, Hwa In; Na, Jin Hee; Jeon, Sangmin; Lim, Seungho; Koo, Heebeom; Han, Sang-Soo; Kang, Sun-Woong; Park, Soon-Jung; Moon, Sung-Hwan; Park, Jae Hyung; Cho, Yong Woo; Kim, Byung-Soo; Kim, Sang Kyoon; Lee, Taekwan; Kim, Dongkyu; Lee, Seulki; Pomper, Martin G; Kwon, Ick Chan; Kim, Kwangmeyung

    2017-09-01

    It is urgently necessary to develop reliable non-invasive stem cell imaging technology for tracking the in vivo fate of transplanted stem cells in living subjects. Herein, we developed a simple and well controlled stem cell imaging method through a combination of metabolic glycoengineering and bioorthogonal copper-free click chemistry. Firstly, the exogenous chemical receptors containing azide (-N 3 ) groups were generated on the surfaces of stem cells through metabolic glycoengineering using metabolic precursor, tetra-acetylated N-azidoacetyl-d-mannosamine(Ac 4 ManNAz). Next, bicyclo[6.1.0]nonyne-modified glycol chitosan nanoparticles (BCN-CNPs) were prepared as imageable nanoparticles to deliver different imaging agents. Cy5.5, iron oxide nanoparticles and gold nanoparticles were conjugated or encapsulated to BCN-CNPs for optical, MR and CT imaging, respectively. These imageable nanoparticles bound chemical receptors on the Ac 4 ManNAz-treated stem cell surface specifically via bioorthogonal copper-free click chemistry. Then they were rapidly taken up by the cell membrane turn-over mechanism resulting in higher endocytic capacity compared non-specific uptake of nanoparticles. During in vivo animal test, BCN-CNP-Cy5.5-labeled stem cells could be continuously tracked by non-invasive optical imaging over 15 days. Furthermore, BCN-CNP-IRON- and BCN-CNP-GOLD-labeled stem cells could be efficiently visualized using in vivo MR and CT imaging demonstrating utility of our stem cell labeling method using chemical receptors. These results conclude that our method based on metabolic glycoengineering and bioorthogonal copper-free click chemistry can stably label stem cells with diverse imageable nanoparticles representing great potential as new stem cell imaging technology. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Study of substrate topographical effects on epithelial cell behavior using etched alpha-particle tracks on PADC films

    Ng, C.K.M.; Poon, W.L.; Li, W.Y.; Cheung, T.; Cheng, S.H.; Yu, K.N.

    2008-01-01

    Micrometer-size pits on the surface of a polymer (polyallyldiglycol carbonate or PADC) substrate created by alpha-particle irradiation and subsequent chemical etching were used to study the topographical effects alone on cell behavior. Vinculin, the cell adhesion and membrane protrusion protein, was used as an indicator of cytoskeletonal reorganization on the substrate and localization of vinculin was used to demonstrate the presence of focal adhesions. In our experiments, vinculin expressed in epithelial HeLa cells cultured on PADC films with track-etch pits, but not in cells cultured on the raw or chemically etched blank films. In other words, vinculin expression was induced by the topography of track-etch pits, while etching of the substrate alone (without alpha-particle irradiation) did not cause up-regulation of vinculin protein expression. HeLa cells cultured on PADC films with track-etch pits also showed changes in cell proliferation, cell area and cell circularity, and were largely contained by the pits. In other words, the cell membrane edges tended to be in contact with the pits. By comparing the correlation between the positions of HeLa cells and the pits, and that between the positions of cells and computer-simulated pits, the tendency for membrane edges of HeLa cells to be in contact with the pits was recognized. This could be explained by inhibition of membrane protrusion at the pits. In conclusion, substrate track-etch pits were an important determinant of epithelial cell behaviors

  11. Assessing the Effectiveness of a Far-Red Fluorescent Reporter for Tracking Stem Cells In Vivo

    Jing Zhou

    2017-12-01

    Full Text Available Far-red fluorescent reporter genes can be used for tracking cells non-invasively in vivo using fluorescence imaging. Here, we investigate the effectiveness of the far-red fluorescent protein, E2-Crimson (E2C, for tracking mouse embryonic cells (mESCs in vivo following subcutaneous administration into mice. Using a knock-in strategy, we introduced E2C into the Rosa26 locus of an E14-Bra-GFP mESC line, and after confirming that the E2C had no obvious effect on the phenotype of the mESCs, we injected them into mice and imaged them over nine days. The results showed that fluorescence intensity was weak, and cells could only be detected when injected at high densities. Furthermore, intensity peaked on day 4 and then started to decrease, despite the fact that tumour volume continued to increase beyond day 4. Histopathological analysis showed that although E2C fluorescence could barely be detected in vivo at day 9, analysis of frozen sections indicated that all mESCs within the tumours continued to express E2C. We hypothesise that the decrease in fluorescence intensity in vivo was probably due to the fact that the mESC tumours became more vascular with time, thus leading to increased absorbance of E2C fluorescence by haemoglobin. We conclude that the E2C reporter has limited use for tracking cells in vivo, at least when introduced as a single copy into the Rosa26 locus.

  12. Coherent optical transients observed in rubidium atomic line filtered Doppler velocimetry experiments

    Fajardo, Mario E., E-mail: mario.fajardo@eglin.af.mil; Molek, Christopher D.; Vesely, Annamaria L. [Air Force Research Laboratory, Munitions Directorate, Ordnance Division, Energetic Materials Branch, AFRL/RWME, 2306 Perimeter Road, Eglin AFB, Florida 32542-5910 (United States)

    2015-10-14

    We report the first successful results from our novel Rubidium Atomic Line Filtered (RALF) Doppler velocimetry apparatus, along with unanticipated oscillatory signals due to coherent optical transients generated within pure Rb vapor cells. RALF is a high-velocity and high-acceleration extension of the well-known Doppler Global Velocimetry (DGV) technique for constructing multi-dimensional flow velocity vector maps in aerodynamics experiments [H. Komine, U.S. Patent No. 4,919,536 (24 April 1990)]. RALF exploits the frequency dependence of pressure-broadened Rb atom optical absorptions in a heated Rb/N{sub 2} gas cell to encode the Doppler shift of reflected near-resonant (λ{sub 0} ≈ 780.24 nm) laser light onto the intensity transmitted by the cell. The present RALF apparatus combines fiber optic and free-space components and was built to determine suitable operating conditions and performance parameters for the Rb/N{sub 2} gas cells. It yields single-spot velocities of thin laser-driven-flyer test surfaces and incorporates a simultaneous Photonic Doppler Velocimetry (PDV) channel [Strand et al., Rev. Sci. Instrum. 77, 083108 (2006)] for validation of the RALF results, which we demonstrate here over the v = 0 to 1 km/s range. Both RALF and DGV presume the vapor cells to be simple Beer's Law optical absorbers, so we were quite surprised to observe oscillatory signals in experiments employing low pressure pure Rb vapor cells. We interpret these oscillations as interference between the Doppler shifted reflected light and the Free Induction Decay (FID) coherent optical transient produced within the pure Rb cells at the original laser frequency; this is confirmed by direct comparison of the PDV and FID signals. We attribute the different behaviors of the Rb/N{sub 2} vs. Rb gas cells to efficient dephasing of the atomic/optical coherences by Rb-N{sub 2} collisions. The minimum necessary N{sub 2} buffer gas density ≈0.3 amagat translates into a

  13. Stereo-particle image velocimetry uncertainty quantification

    Bhattacharya, Sayantan; Vlachos, Pavlos P; Charonko, John J

    2017-01-01

    Particle image velocimetry (PIV) measurements are subject to multiple elemental error sources and thus estimating overall measurement uncertainty is challenging. Recent advances have led to a posteriori uncertainty estimation methods for planar two-component PIV. However, no complete methodology exists for uncertainty quantification in stereo PIV. In the current work, a comprehensive framework is presented to quantify the uncertainty stemming from stereo registration error and combine it with the underlying planar velocity uncertainties. The disparity in particle locations of the dewarped images is used to estimate the positional uncertainty of the world coordinate system, which is then propagated to the uncertainty in the calibration mapping function coefficients. Next, the calibration uncertainty is combined with the planar uncertainty fields of the individual cameras through an uncertainty propagation equation and uncertainty estimates are obtained for all three velocity components. The methodology was tested with synthetic stereo PIV data for different light sheet thicknesses, with and without registration error, and also validated with an experimental vortex ring case from 2014 PIV challenge. Thorough sensitivity analysis was performed to assess the relative impact of the various parameters to the overall uncertainty. The results suggest that in absence of any disparity, the stereo PIV uncertainty prediction method is more sensitive to the planar uncertainty estimates than to the angle uncertainty, although the latter is not negligible for non-zero disparity. Overall the presented uncertainty quantification framework showed excellent agreement between the error and uncertainty RMS values for both the synthetic and the experimental data and demonstrated reliable uncertainty prediction coverage. This stereo PIV uncertainty quantification framework provides the first comprehensive treatment on the subject and potentially lays foundations applicable to volumetric

  14. Real-time tracking of cell cycle progression during CD8+ effector and memory T-cell differentiation.

    Kinjyo, Ichiko; Qin, Jim; Tan, Sioh-Yang; Wellard, Cameron J; Mrass, Paulus; Ritchie, William; Doi, Atsushi; Cavanagh, Lois L; Tomura, Michio; Sakaue-Sawano, Asako; Kanagawa, Osami; Miyawaki, Atsushi; Hodgkin, Philip D; Weninger, Wolfgang

    2015-02-24

    The precise pathways of memory T-cell differentiation are incompletely understood. Here we exploit transgenic mice expressing fluorescent cell cycle indicators to longitudinally track the division dynamics of individual CD8(+) T cells. During influenza virus infection in vivo, naive T cells enter a CD62L(intermediate) state of fast proliferation, which continues for at least nine generations. At the peak of the anti-viral immune response, a subpopulation of these cells markedly reduces their cycling speed and acquires a CD62L(hi) central memory cell phenotype. Construction of T-cell family division trees in vitro reveals two patterns of proliferation dynamics. While cells initially divide rapidly with moderate stochastic variations of cycling times after each generation, a slow-cycling subpopulation displaying a CD62L(hi) memory phenotype appears after eight divisions. Phenotype and cell cycle duration are inherited by the progeny of slow cyclers. We propose that memory precursors cell-intrinsically modulate their proliferative activity to diversify differentiation pathways.

  15. Quantitative Cell Cycle Analysis Based on an Endogenous All-in-One Reporter for Cell Tracking and Classification

    Thomas Zerjatke

    2017-05-01

    Full Text Available Cell cycle kinetics are crucial to cell fate decisions. Although live imaging has provided extensive insights into this relationship at the single-cell level, the limited number of fluorescent markers that can be used in a single experiment has hindered efforts to link the dynamics of individual proteins responsible for decision making directly to cell cycle progression. Here, we present fluorescently tagged endogenous proliferating cell nuclear antigen (PCNA as an all-in-one cell cycle reporter that allows simultaneous analysis of cell cycle progression, including the transition into quiescence, and the dynamics of individual fate determinants. We also provide an image analysis pipeline for automated segmentation, tracking, and classification of all cell cycle phases. Combining the all-in-one reporter with labeled endogenous cyclin D1 and p21 as prime examples of cell-cycle-regulated fate determinants, we show how cell cycle and quantitative protein dynamics can be simultaneously extracted to gain insights into G1 phase regulation and responses to perturbations.

  16. Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice

    Papaioannou Virginia E

    2004-12-01

    Full Text Available Abstract Background Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse transgenic and targeted mutagenesis applications. Results We have used embryonic stem cell-mediated transgenesis to label cells at sub-cellular resolution in vivo, and to evaluate fusion of a human histone protein to green fluorescent protein for ubiquitous fluorescent labeling of nucleosomes in mice. To this end we have generated embryonic stem cells and a corresponding strain of mice that is viable and fertile and exhibits widespread chromatin-localized reporter expression. High levels of transgene expression are maintained in a constitutive manner. Viability and fertility of homozygous transgenic animals demonstrates that this reporter is developmentally neutral and does not interfere with mitosis or meiosis. Conclusions Using various optical imaging modalities including wide-field, spinning disc confocal, and laser scanning confocal and multiphoton excitation microscopy, we can identify cells in various stages of the cell cycle. We can identify cells in interphase, cells undergoing mitosis or cell death. We demonstrate that this histone fusion reporter allows the direct visualization of active chromatin in situ. Since this reporter segments three-dimensional space, it permits the visualization of individual cells within a population, and so facilitates tracking cell position over time. It is therefore attractive for use in multidimensional studies of in vivo cell behavior and cell fate.

  17. Cellular transfer of magnetic nanoparticles via cell microvesicles: impact on cell tracking by magnetic resonance imaging.

    Silva, Amanda K Andriola; Wilhelm, Claire; Kolosnjaj-Tabi, Jelena; Luciani, Nathalie; Gazeau, Florence

    2012-05-01

    Cell labeling with magnetic nanoparticles can be used to monitor the fate of transplanted cells in vivo by magnetic resonance imaging. However, nanoparticles initially internalized in administered cells might end up in other cells of the host organism. We investigated a mechanism of intercellular cross-transfer of magnetic nanoparticles to different types of recipient cells via cell microvesicles released under cellular stress. Three cell types (mesenchymal stem cells, endothelial cells and macrophages) were labeled with 8-nm iron oxide nanoparticles. Then cells underwent starvation stress, during which they produced microvesicles that were subsequently transferred to unlabeled recipient cells. The analysis of the magnetophoretic mobility of donor cells indicated that magnetic load was partially lost under cell stress. Microvesicles shed by stressed cells participated in the release of magnetic label. Moreover, such microvesicles were uptaken by naïve cells, resulting in cellular redistribution of nanoparticles. Iron load of recipient cells allowed their detection by MRI. Cell microvesicles released under stress may be disseminated throughout the organism, where they can be uptaken by host cells. The transferred cargo may be sufficient to allow MRI detection of these secondarily labeled cells, leading to misinterpretations of the effectiveness of transplanted cells.

  18. Magneto-optical labeling of fetal neural stem cells for in vivo MRI tracking.

    Flexman, J A; Minoshima, S; Kim, Y; Cross, D J

    2006-01-01

    Neural stem cell therapy for neurological pathologies, such as Alzheimer's and Parkinson's disease, may delay the onset of symptoms, replace damaged neurons and/or support the survival of endogenous cells. Magnetic resonance imaging (MRI) can be used to track magnetically labeled cells in vivo to observe migration. Prior to transplantation, labeled cells must be characterized to show that they retain their intrinsic properties, such as cell proliferation into neurospheres in a supplemented environment. In vivo images must also be correlated to sensitive, histological markers. In this study, we show that fetus-derived neural stem cells can be co-labeled with superparamagnetic iron oxide and PKH26, a fluorescent dye. Labeled cells retain the ability to proliferate into neurospheres in culture, but labeling prevents neurospheres from merging in a non-adherent culture environment. After labeled NSCs were transplantation into the rat brain, their location and subsequent migration along the corpus callosum was detected using MRI. This study demonstrates an imaging paradigm with which to develop an in vivo assay for quantitatively evaluating fetal neural stem cell migration.

  19. Polarimetric radar convective cell tracking reveals large sensitivity of cloud precipitation and electrification properties to CCN

    Hu, J.; Rosenfeld, D.; Zhang, P.; Snyder, J.; Orville, R. E.; Ryzhkov, A.; Zrnic, D.; Williams, E. R.; Zhang, R.

    2017-12-01

    Here we apply the cell tracking methodology, shown in our companion poster, to quantifying factors affecting the vigor and the time-height evolution of hydrometeors and electrification properties of convective cells. Benefitting from the Dual-polarimetric NEXRAD radar network, we composite more than 5000 well-tracked cells among three radars (at Houston, Lubbock and Oklahoma City), stratified by CCN, CAPE and land/sea locations. The analyzed cell properties include Z, ZDR, Kdp, and ρhv, Dm (raindrop diameter) and Nw (raindrop concentration) by the algorithm of Bringi et al. (2003). Lightning Mapping Array (LMA) data is also included in the analysis, which provides a 3D structure of lightning occurrence and RF power. The contrasting CCN conditions over marine, land, pristine and polluted areas are identified based on the satellite retrieval technique described in Rosenfeld et al. (2016). The results show that more CCN are associated with: Increased echo top height, manifesting the invigoration effect. Enhanced reflectivities, especially above the freezing level at around 4.5 km. Raindrop sizes at the initial stage increase at the expense of their concentrations, due to the smaller cloud droplets and suppressed coalescence. Larger propensity for hail. Lightning sources increase with greater CCN concentration and is likely due to the delayed warm rain process and enhanced mixed phase process under more CCN condition, when activated CCN into cloud droplets is too high (> 1000 cm-3) the glaciation is delayed too much and leave little ice at lower levels and thus decrease lightning activity. Land pristine clouds have fewer lightning sources than polluted clouds. Marine pristine clouds seldom have lightning Increased CAPE had a similar effect to the effect of added CCN. The cloud tracking and properties are obtained by a new methodology of Multi-Cell Identification and Tracking (MCIT) algorithm (Hu et al, 2017), with details about the algorithm to be found in the author

  20. Cre/lox-assisted non-invasive in vivo tracking of specific cell populations by positron emission tomography.

    Thunemann, Martin; Schörg, Barbara F; Feil, Susanne; Lin, Yun; Voelkl, Jakob; Golla, Matthias; Vachaviolos, Angelos; Kohlhofer, Ursula; Quintanilla-Martinez, Leticia; Olbrich, Marcus; Ehrlichmann, Walter; Reischl, Gerald; Griessinger, Christoph M; Langer, Harald F; Gawaz, Meinrad; Lang, Florian; Schäfers, Michael; Kneilling, Manfred; Pichler, Bernd J; Feil, Robert

    2017-09-05

    Many pathophysiological processes are associated with proliferation, migration or death of distinct cell populations. Monitoring specific cell types and their progeny in a non-invasive, longitudinal and quantitative manner is still challenging. Here we show a novel cell-tracking system that combines Cre/lox-assisted cell fate mapping with a thymidine kinase (sr39tk) reporter gene for cell detection by positron emission tomography (PET). We generate Rosa26-mT/sr39tk PET reporter mice and induce sr39tk expression in platelets, T lymphocytes or cardiomyocytes. As proof of concept, we demonstrate that our mouse model permits longitudinal PET imaging and quantification of T-cell homing during inflammation and cardiomyocyte viability after myocardial infarction. Moreover, Rosa26-mT/sr39tk mice are useful for whole-body characterization of transgenic Cre mice and to detect previously unknown Cre activity. We anticipate that the Cre-switchable PET reporter mice will be broadly applicable for non-invasive long-term tracking of selected cell populations in vivo.Non-invasive cell tracking is a powerful method to visualize cells in vivo under physiological and pathophysiological conditions. Here Thunemann et al. generate a mouse model for in vivo tracking and quantification of specific cell types by combining a PET reporter gene with Cre-dependent activation that can be exploited for any cell population for which a Cre mouse line is available.

  1. Comparison of Simultaneous PIV and Hydroxyl Tagging Velocimetry in Low Velocity Flows

    Andre, Matthieu A.; Bardet, Philippe M.; Burns, Ross A.; Danehy, Paul M.

    2016-01-01

    Hydroxyl tagging velocimetry (HTV) is a molecular tagging velocimetry (MTV) technique that relies on the photo- dissociation of water vapor into OH radicals and their subsequent tracking using laser-induced fluorescence. At ambient temperature in air, the OH species lifetime is about 50 micro-s. The feasibility of using HTV for probing low- speed flows (a few m/s) is investigated by using an inert, heated gas as a means to increase the OH species lifetime. Unlike particle-based techniques, MTV does not suffer from tracer settling, which is particularly problematic at low speeds. Furthermore, the flow needs to be seeded with only a small mole fraction of water vapor, making it safer for both the user and facilities than other MTV techniques based on corrosive or toxic chemical tracers. HTV is demonstrated on a steam-seeded nitrogen jet at approximately 75 C in the laminar (Umean=3.31 m/s, Re=1,540), transitional (Umean=4.48 m/s, Re=2,039), and turbulent (Umean=6.91 m/s, Re=3,016) regimes at atmospheric pressure. The measured velocity profiles are compared with particle image velocimetry (PIV) measurements performed simultaneously with a second imager. Seeding for the PIV is achieved by introducing micron-sized water droplets into the flow with the steam; the same laser sheet is used for PIV and HTV to guarantee spatial and temporal overlap of the data. Optimizing each of these methods, however, requires conflicting operating conditions: higher temperatures benefit the HTV signals but reduce the available seed density for the PIV through evaporation. Nevertheless, data are found to agree within 10% for the instantaneous velocity profiles and within 5% for the mean profiles and demonstrate the feasibility of HTV for low-speed flows at moderate to high temperatures.

  2. Magnetic cell labeling of primary and stem cell-derived pig hepatocytes for MRI-based cell tracking of hepatocyte transplantation.

    Dwayne R Roach

    Full Text Available Pig hepatocytes are an important investigational tool for optimizing hepatocyte transplantation schemes in both allogeneic and xenogeneic transplant scenarios. MRI can be used to serially monitor the transplanted cells, but only if the hepatocytes can be labeled with a magnetic particle. In this work, we describe culture conditions for magnetic cell labeling of cells from two different pig hepatocyte cell sources; primary pig hepatocytes (ppHEP and stem cell-derived hepatocytes (PICM-19FF. The magnetic particle is a micron-sized iron oxide particle (MPIO that has been extensively studied for magnetic cell labeling for MRI-based cell tracking. ppHEP could endocytose MPIO with labeling percentages as high as 70%, achieving iron content as high as ~55 pg/cell, with >75% viability. PICM-19FF had labeling >97%, achieving iron content ~38 pg/cell, with viability >99%. Extensive morphological and functional assays indicated that magnetic cell labeling was benign to the cells. The results encourage the use of MRI-based cell tracking for the development and clinical use of hepatocyte transplantation methodologies. Further, these results generally highlight the importance of functional cell assays in the evaluation of contrast agent biocompatibility.

  3. Synthetic and biogenic magnetite nanoparticles for tracking of stem cells and dendritic cells

    Schwarz, Sebastian; Fernandes, Fabiana; Sanroman, Laura; Hodenius, Michael; Lang, Claus; Himmelreich, Uwe; Schmitz-Rode, Thomas; Schueler, Dirk; Hoehn, Mathias

    2009-01-01

    Accurate delivery of cells to target organs is critical for success of cell-based therapies with stem cells or immune cells such as antigen-presenting dendritic cells (DC). Labeling with contrast agents before implantation provides a powerful means for monitoring cellular migration using magnetic resonance imaging (MRI). In this study, we investigated the uptake of fully synthesized or bacterial magnetic nanoparticles (MNPs) into hematopoietic Flt3 + stem cells and DC from mouse bone marrow. We show that (i) uptake of both synthetic and biogenic nanoparticles into cells endow magnetic activity and (ii) low numbers of MNP-loaded cells are readily detected by MRI.

  4. Blood on the tracks: hematopoietic stem cell-endothelial cell interactions in homing and engraftment.

    Perlin, Julie R; Sporrij, Audrey; Zon, Leonard I

    2017-08-01

    Cells of the hematopoietic system undergo rapid turnover. Each day, humans require the production of about one hundred billion new blood cells for proper function. Hematopoietic stem cells (HSCs) are rare cells that reside in specialized niches and are required throughout life to produce specific progenitor cells that will replenish all blood lineages. There is, however, an incomplete understanding of the molecular and physical properties that regulate HSC migration, homing, engraftment, and maintenance in the niche. Endothelial cells (ECs) are intimately associated with HSCs throughout the life of the stem cell, from the specialized endothelial cells that give rise to HSCs, to the perivascular niche endothelial cells that regulate HSC homeostasis. Recent studies have dissected the unique molecular and physical properties of the endothelial cells in the HSC vascular niche and their role in HSC biology, which may be manipulated to enhance hematopoietic stem cell transplantation therapies.

  5. Operationalizing Civilian Protection in Mali: The Case for a Civilian Casualty Tracking, Analysis, and Response Cell

    Marla B. Keenan

    2013-06-01

    Full Text Available This practice note details an emerging best practice of civilian harm mitigation in armed conflict: namely, the creation of civilian casualty tracking, analysis and response processes by a warring party or peace operation force. It asserts that in Iraq, Afghanistan and soon Somalia, these processes to better understand civilian harm and address consequences have positively shaped mission tactics, training, and overall operations. In both Iraq and Afghanistan, tracking and analysis has lead to a marked decrease in civilian casualties and facilitated the making of amends for any civilian losses. The paper argues that for warring parties to achieve their mission—particularly one with a protection of civilians mandate as with the United Nations Multidimensional Integrated Stabilization Mission in Mali (MINUSMA—they must fully understand the impact of their actions on the civilian population, positive or negative. For this reason, a Civilian Casualty Tracking, Analysis, and Response Cell should be created for MINUSMA to improve its ability mitigate risk to civilians as required by its Security Council mandate.

  6. Tracking virus-specific CD4+ T cells during and after acute hepatitis C virus infection.

    Michaela Lucas

    2007-07-01

    Full Text Available CD4+ T cell help is critical in maintaining antiviral immune responses and such help has been shown to be sustained in acute resolving hepatitis C. In contrast, in evolving chronic hepatitis C CD4+ T cell helper responses appear to be absent or short-lived, using functional assays.Here we used a novel HLA-DR1 tetramer containing a highly targeted CD4+ T cell epitope from the hepatitis C virus non-structural protein 4 to track number and phenotype of hepatitis C virus specific CD4+ T cells in a cohort of seven HLA-DR1 positive patients with acute hepatitis C in comparison to patients with chronic or resolved hepatitis C. We observed peptide-specific T cells in all seven patients with acute hepatitis C regardless of outcome at frequencies up to 0.65% of CD4+ T cells. Among patients who transiently controlled virus replication we observed loss of function, and/or physical deletion of tetramer+ CD4+ T cells before viral recrudescence. In some patients with chronic hepatitis C very low numbers of tetramer+ cells were detectable in peripheral blood, compared to robust responses detected in spontaneous resolvers. Importantly we did not observe escape mutations in this key CD4+ T cell epitope in patients with evolving chronic hepatitis C.During acute hepatitis C a CD4+ T cell response against this epitope is readily induced in most, if not all, HLA-DR1+ patients. This antiviral T cell population becomes functionally impaired or is deleted early in the course of disease in those where viremia persists.

  7. Tracking of adipose tissue-derived progenitor cells using two magnetic nanoparticle types

    Kasten, Annika; Siegmund, Birte J. [Department of Oral and Maxillofacial Surgery, Facial Plastic Surgery, Rostock University Medical Center, Schillingallee 35 D-18057 Rostock (Germany); Grüttner, Cordula [Micromod Partikeltechnologie GmbH, Warnemünde, D-18115 Rostock (Germany); Kühn, Jens-Peter [Department of Radiology and Neuroradiology, Greifswald University Medical Center, D-17475 Greifswald (Germany); Frerich, Bernhard, E-mail: bernhard.frerich@med.uni-rostock.de [Department of Oral and Maxillofacial Surgery, Facial Plastic Surgery, Rostock University Medical Center, Schillingallee 35 D-18057 Rostock (Germany)

    2015-04-15

    Magnetic resonance imaging (MRI) is to be considered as an emerging detection technique for cell tracking experiments to evaluate the fate of transplanted progenitor cells and develop successful cell therapies for tissue engineering. Adipose tissue engineering using adipose tissue-derived progenitor cells has been advocated for the cure of soft tissue defects or for persistent soft tissue augmentation. Adipose tissue-derived progenitor cells were differentiated into the adipogenic lineage and labeled with two different types of magnetic iron oxide nanoparticles in varying concentrations which resulted in a concentration-dependent reduction of gene expression of adipogenic differentiation markers, adiponectin and fatty acid-binding protein 4 (FABP4), whereas the metabolic activity was not altered. As a result, only low nanoparticle concentrations for labeling were used for in vivo experiments. Cells were seeded onto collagen scaffolds and subcutaneously implanted into severe combined immunodeficient (SCID) mice. At 24 h as well as 28 days after implantation, MRI analyses were performed visualizing nanoparticle-labeled cells using T2-weighted sequences. The quantification of absolute volume of the scaffolds revealed a decrease of volume over time in all experimental groups. The distribution of nanoparticle-labeled cells within the scaffolds varied likewise over time. - Highlights: • Adipose tissue-derived stem cells (ASC) were labeled with magnetic iron oxide nanoparticles. • Nanoparticles influenced the adipogenic differentiation of ASC. • Labeled cells were seeded onto collagen scaffolds and implanted in SCID mice. • Nanoparticle-labeled cells were visualized in vivo using T2-weighted sequences. • Volume of collagen scaffolds was decreased over time after implantation.

  8. Tracking of adipose tissue-derived progenitor cells using two magnetic nanoparticle types

    Kasten, Annika; Siegmund, Birte J.; Grüttner, Cordula; Kühn, Jens-Peter; Frerich, Bernhard

    2015-01-01

    Magnetic resonance imaging (MRI) is to be considered as an emerging detection technique for cell tracking experiments to evaluate the fate of transplanted progenitor cells and develop successful cell therapies for tissue engineering. Adipose tissue engineering using adipose tissue-derived progenitor cells has been advocated for the cure of soft tissue defects or for persistent soft tissue augmentation. Adipose tissue-derived progenitor cells were differentiated into the adipogenic lineage and labeled with two different types of magnetic iron oxide nanoparticles in varying concentrations which resulted in a concentration-dependent reduction of gene expression of adipogenic differentiation markers, adiponectin and fatty acid-binding protein 4 (FABP4), whereas the metabolic activity was not altered. As a result, only low nanoparticle concentrations for labeling were used for in vivo experiments. Cells were seeded onto collagen scaffolds and subcutaneously implanted into severe combined immunodeficient (SCID) mice. At 24 h as well as 28 days after implantation, MRI analyses were performed visualizing nanoparticle-labeled cells using T2-weighted sequences. The quantification of absolute volume of the scaffolds revealed a decrease of volume over time in all experimental groups. The distribution of nanoparticle-labeled cells within the scaffolds varied likewise over time. - Highlights: • Adipose tissue-derived stem cells (ASC) were labeled with magnetic iron oxide nanoparticles. • Nanoparticles influenced the adipogenic differentiation of ASC. • Labeled cells were seeded onto collagen scaffolds and implanted in SCID mice. • Nanoparticle-labeled cells were visualized in vivo using T2-weighted sequences. • Volume of collagen scaffolds was decreased over time after implantation

  9. Performing particle image velocimetry using artificial neural networks: a proof-of-concept

    Rabault, Jean; Kolaas, Jostein; Jensen, Atle

    2017-12-01

    Traditional programs based on feature engineering are underperforming on a steadily increasing number of tasks compared with artificial neural networks (ANNs), in particular for image analysis. Image analysis is widely used in fluid mechanics when performing particle image velocimetry (PIV) and particle tracking velocimetry (PTV), and therefore it is natural to test the ability of ANNs to perform such tasks. We report for the first time the use of convolutional neural networks (CNNs) and fully connected neural networks (FCNNs) for performing end-to-end PIV. Realistic synthetic images are used for training the networks and several synthetic test cases are used to assess the quality of each network’s predictions and compare them with state-of-the-art PIV software. In addition, we present tests on real-world data that prove ANNs can be used not only with synthetic images but also with more noisy, imperfect images obtained in a real experimental setup. While the ANNs we present have slightly higher root mean square error than state-of-the-art cross-correlation methods, they perform better near edges and allow for higher spatial resolution than such methods. In addition, it is likely that one could with further work develop ANNs which perform better that the proof-of-concept we offer.

  10. Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics.

    Burke, Russell T; Orth, James D

    2016-05-14

    The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing factor in the overall outcome. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. These methods are important, but they have several shortcomings. Variability in drug responses between cancer and normal cells, and between cells of different cancer origin, and transient and rare responses are difficult to understand using population averaging assays and without being able to directly track and analyze them longitudinally. The microscope is particularly well suited to image live cells. Advancements in technology enable us to routinely image cells at a resolution that enables not only cell tracking, but also the observation of a variety of cellular responses. We describe an approach in detail that allows for the continuous time-lapse imaging of cells during the drug response for essentially as long as desired, typically up to 96 hr. Using variations of the approach, cells can be monitored for weeks. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. We also discuss variations of the technique and its flexibility, and highlight some common pitfalls.

  11. Tracking chemical changes in a live cell: Biomedical applications of SR-FTIR spectromicroscopy

    Holman, Hoi-Ying N.; Martin, Michael C.; McKinney, Wayne R.

    2002-07-25

    Synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectromicroscopy is a newly emerging bioanalytical and imaging tool. This unique technique provides mid-infrared (IR) spectra, hence chemical information, with high signal-to-noise at spatial resolutions as fine as 3 to 10 microns. Thus it enables researchers to locate, identify, and track specific chemical events within an individual living mammalian cell. Mid-IR photons are too low in energy (0.05 - 0.5 eV) to either break bonds or to cause ionization. In this review, we show that the synchrotron IR beam has no detectable effects on the short- and long-term viability, reproductive integrity, cell-cycle progression, and mitochondrial metabolism in living human cells, and produces only minimal sample heating (< 0.5 degrees C). We will then present several examples demonstrating the application potentials of SR-FTIR spectromicroscopy in biomedical research. These will include monitoring living cells progressing through the cell cycle, including death, and cells reacting to dilute concentrations of toxins.

  12. Tracking chemical changes in a live cell: Biomedical applications of SR-FTIR spectromicroscopy

    Holman, Hoi-Ying N.; Martin, Michael C.; McKinney, Wayne R.

    2002-01-01

    Synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectromicroscopy is a newly emerging bioanalytical and imaging tool. This unique technique provides mid-infrared (IR) spectra, hence chemical information, with high signal-to-noise at spatial resolutions as fine as 3 to 10 microns. Thus it enables researchers to locate, identify, and track specific chemical events within an individual living mammalian cell. Mid-IR photons are too low in energy (0.05 - 0.5 eV) to either break bonds or to cause ionization. In this review, we show that the synchrotron IR beam has no detectable effects on the short- and long-term viability, reproductive integrity, cell-cycle progression, and mitochondrial metabolism in living human cells, and produces only minimal sample heating (< 0.5 degrees C). We will then present several examples demonstrating the application potentials of SR-FTIR spectromicroscopy in biomedical research. These will include monitoring living cells progressing through the cell cycle, including death, and cells reacting to dilute concentrations of toxins

  13. Swarming and complex pattern formation in Paenibacillus vortex studied by imaging and tracking cells

    Jacob Eshel

    2008-02-01

    Full Text Available Abstract Background Swarming motility allows microorganisms to move rapidly over surfaces. The Gram-positive bacterium Paenibacillus vortex exhibits advanced cooperative motility on agar plates resulting in intricate colonial patterns with geometries that are highly sensitive to the environment. The cellular mechanisms that underpin the complex multicellular organization of such a simple organism are not well understood. Results Swarming by P. vortex was studied by real-time light microscopy, by in situ scanning electron microscopy and by tracking the spread of antibiotic-resistant cells within antibiotic-sensitive colonies. When swarming, P. vortex was found to be peritrichously flagellated. Swarming by the curved cells of P. vortex occurred on an extremely wide range of media and agar concentrations (0.3 to 2.2% w/v. At high agar concentrations (> 1% w/v rotating colonies formed that could be detached from the main mass of cells by withdrawal of cells into the latter. On lower percentage agars, cells moved in an extended network composed of interconnected "snakes" with short-term collision avoidance and sensitivity to extracts from swarming cells. P. vortex formed single Petri dish-wide "supercolonies" with a colony-wide exchange of motile cells. Swarming cells were coupled by rapidly forming, reversible and non-rigid connections to form a loose raft, apparently connected via flagella. Inhibitors of swarming (p-Nitrophenylglycerol and Congo Red were identified. Mitomycin C was used to trigger filamentation without inhibiting growth or swarming; this facilitated dissection of the detail of swarming. Mitomycin C treatment resulted in malcoordinated swarming and abortive side branch formation and a strong tendency by a subpopulation of the cells to form minimal rotating aggregates of only a few cells. Conclusion P. vortex creates complex macroscopic colonies within which there is considerable reflux and movement and interaction of cells. Cell

  14. Comparison of Pyranometers and Reference Cells on Fixed and One-Axis Tracking Surfaces: Preprint

    Dooraghi, Michael R [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Sengupta, Manajit [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Vignola, Frank [University of Oregon; Peterson, Josh [University of Oregon; Mavromatakis, Fotis [Technological Educational Institute of Crete; Chiu, Chun-Yu [University of Oregon

    2017-12-19

    A wide variety of sensors are used to monitor the irradiance incident on solar modules to evaluate the performance of photovoltaic (PV) systems. These instruments range from secondary standard pyranometers to photodiode-based pyranometers to reference cells. Although instruments are mounted in the plane of array of the modules, a wide range of results have been obtained. Some of these difference have been assumed to come from systematic uncertainties associated with the irradiance sensors. This study is an attempt to quantify these differences by comparing the output of selected thermopile pyranometers to photodiode-based pyranometers and reference cells on a horizontal surface, a fixed-tilt surface, and a one-axis tracking surface. This analysis focuses on clear-sky results from two sites with different climatic conditions. Several important features were observed. Photodiode-based pyranometers and reference cells produce widely different results under clear skies, especially at larger angles of incidence, even though both instruments are based on measuring the short-circuit current of solar cells. The difference is caused by the scattering of light as it passes through the glazing of the reference cell or the diffuser lens of the photodioded-base pyranometer. Both instruments are shown to have similar response to the spectral distribution of the irradiance when compared to the thermopile-based pyranometer, which has a response nearly independent of the wavelength of light used by PV modules.

  15. Tracking of plus-ends reveals microtubule functional diversity in different cell types

    Shaebani, M. Reza; Pasula, Aravind; Ott, Albrecht; Santen, Ludger

    2016-07-01

    Many cellular processes are tightly connected to the dynamics of microtubules (MTs). While in neuronal axons MTs mainly regulate intracellular trafficking, they participate in cytoskeleton reorganization in many other eukaryotic cells, enabling the cell to efficiently adapt to changes in the environment. We show that the functional differences of MTs in different cell types and regions is reflected in the dynamic properties of MT tips. Using plus-end tracking proteins EB1 to monitor growing MT plus-ends, we show that MT dynamics and life cycle in axons of human neurons significantly differ from that of fibroblast cells. The density of plus-ends, as well as the rescue and catastrophe frequencies increase while the growth rate decreases toward the fibroblast cell margin. This results in a rather stable filamentous network structure and maintains the connection between nucleus and membrane. In contrast, plus-ends are uniformly distributed along the axons and exhibit diverse polymerization run times and spatially homogeneous rescue and catastrophe frequencies, leading to MT segments of various lengths. The probability distributions of the excursion length of polymerization and the MT length both follow nearly exponential tails, in agreement with the analytical predictions of a two-state model of MT dynamics.

  16. Tracking Cell Surface GABAB Receptors Using an α-Bungarotoxin Tag*

    Wilkins, Megan E.; Li, Xinyan; Smart, Trevor G.

    2008-01-01

    GABAB receptors mediate slow synaptic inhibition in the central nervous system and are important for synaptic plasticity as well as being implicated in disease. Located at pre- and postsynaptic sites, GABAB receptors will influence cell excitability, but their effectiveness in doing so will be dependent, in part, on their trafficking to, and stability on, the cell surface membrane. To examine the dynamic behavior of GABAB receptors in GIRK cells and neurons, we have devised a method that is based on tagging the receptor with the binding site components for the neurotoxin, α-bungarotoxin. By using the α-bungarotoxin binding site-tagged GABAB R1a subunit (R1aBBS), co-expressed with the R2 subunit, we can track receptor mobility using the small reporter, α-bungarotoxin-conjugated rhodamine. In this way, the rates of internalization and membrane insertion for these receptors could be measured with fixed and live cells. The results indicate that GABAB receptors rapidly turnover in the cell membrane, with the rate of internalization affected by the state of receptor activation. The bungarotoxin-based method of receptor-tagging seems ideally suited to follow the dynamic regulation of other G-protein-coupled receptors. PMID:18812318

  17. Tracking cell surface GABAB receptors using an alpha-bungarotoxin tag.

    Wilkins, Megan E; Li, Xinyan; Smart, Trevor G

    2008-12-12

    GABA(B) receptors mediate slow synaptic inhibition in the central nervous system and are important for synaptic plasticity as well as being implicated in disease. Located at pre- and postsynaptic sites, GABA(B) receptors will influence cell excitability, but their effectiveness in doing so will be dependent, in part, on their trafficking to, and stability on, the cell surface membrane. To examine the dynamic behavior of GABA(B) receptors in GIRK cells and neurons, we have devised a method that is based on tagging the receptor with the binding site components for the neurotoxin, alpha-bungarotoxin. By using the alpha-bungarotoxin binding site-tagged GABA(B) R1a subunit (R1a(BBS)), co-expressed with the R2 subunit, we can track receptor mobility using the small reporter, alpha-bungarotoxin-conjugated rhodamine. In this way, the rates of internalization and membrane insertion for these receptors could be measured with fixed and live cells. The results indicate that GABA(B) receptors rapidly turnover in the cell membrane, with the rate of internalization affected by the state of receptor activation. The bungarotoxin-based method of receptor-tagging seems ideally suited to follow the dynamic regulation of other G-protein-coupled receptors.

  18. Comparison of Pyranometers and Reference Cells on Fixed and One-axis Tracking Surfaces

    Dooraghi, Michael R [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Sengupta, Manajit [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Vignola, Frank [University of Oregon; Peterson, Josh [University of Oregon; Mavromatakis, Fotis [Technological Educational Institute of Crete; Chiu, Chun-Yu [University of Oregon

    2017-10-12

    Photovoltaic (PV) system perfomance is monitored by a wide variety of sensors. These instruments range from secondary standard pyranometers to photodiode-based pyranometers to reference cells. Although instruments are mounted in the plane of array of the modules a wide range of results have been obtained. Some of these difference have been assumed to come from systematic uncertainties associated with the irradiance sensors. This study is an attempt to quantify these differences by comparing the output of selected thermopile-based pyranometers to photodiode-based pyranometers and reference cells on a horizontal surface, a fixed-tilt surface, and a one-axis tracking surface. This analysis focuses on clear-sky results from two sites with different climatic conditions. Several important features were observed. Photodiode-based pyranometers and reference cells produce widely different results under clear skies, especially at larger angles-of-incidence even though both instruments are based on measuring the short circuit current of solar cells. The difference is caused by the scattering of light as it passes through the glazing of the reference cell or the diffuser lens of the photodioded- base pyranometer. Both instruments are shown to have similar response to the spectral distribution of the irradiance when compared to the thermopile-based pyranometer that has a response nearly independent of the wavelength of light used by PV modules.

  19. Uncovering homo-and hetero-interactions on the cell membrane using single particle tracking approaches

    Torreno-Pina, Juan A; Manzo, Carlo; Garcia-Parajo, Maria F

    2016-01-01

    The plasma membrane of eukaryotic cells is responsible for a myriad of functions that regulate cell physiology and plays a crucial role in a multitude of processes that include adhesion, migration, signaling recognition and cell–cell communication. This is accomplished by specific interactions between different membrane components such as lipids and proteins on the lipid bilayer but also through interactions with the underlying cortical actin cytoskeleton on the intracellular side and the glycocalyx matrix in close proximity to the extracellular side. Advanced biophysical techniques, including single particle tracking (SPT) have revealed that the lateral diffusion of molecular components on the plasma membrane represents a landmark manifestation of such interactions. Indeed, by studying changes in the diffusivity of individual membrane molecules, including sub-diffusion, confined diffusion and/or transient arrest of molecules in membrane compartments, it has been possible to gain insight on the nature of molecular interactions and to infer on its functional role for cell response. In this review, we will revise some exciting results where SPT has been crucial to reveal homo- and hetero-interactions on the cell membrane. (paper)

  20. Characterization of exosomes derived from ovarian cancer cells and normal ovarian epithelial cells by nanoparticle tracking analysis.

    Zhang, Wei; Peng, Peng; Kuang, Yun; Yang, Jiaxin; Cao, Dongyan; You, Yan; Shen, Keng

    2016-03-01

    Cellular exosomes are involved in many disease processes and have the potential to be used for diagnosis and treatment. In this study, we compared the characteristics of exosomes derived from human ovarian epithelial cells (HOSEPiC) and three epithelial ovarian cancer cell lines (OVCAR3, IGROV1, and ES-2) to investigate the differences between exosomes originating from normal and malignant cells. Two established colloid-chemical methodologies, electron microscopy (EM) and dynamic light scattering (DLS), and a relatively new method, nanoparticle tracking analysis (NTA), were used to measure the size and size distribution of exosomes. The concentration and epithelial cellular adhesion molecule (EpCAM) expression of exosomes were measured by NTA. Quantum dots were conjugated with anti-EpCAM to label exosomes, and the labeled exosomes were detected by NTA in fluorescent mode. The normal-cell-derived exosomes were significantly larger than those derived from malignant cells, and exosomes were successfully labeled using anti-EpCAM-conjugated quantum dots. Exosomes from different cell lines may vary in size, and exosomes might be considered as potential diagnosis biomarkers. NTA can be considered a useful, efficient, and objective method for the study of different exosomes and their unique properties in ovarian cancer.

  1. Tracking fusion of human mesenchymal stem cells after transplantation to the heart.

    Freeman, Brian T; Kouris, Nicholas A; Ogle, Brenda M

    2015-06-01

    Evidence suggests that transplanted mesenchymal stem cells (MSCs) can aid recovery of damaged myocardium caused by myocardial infarction. One possible mechanism for MSC-mediated recovery is reprogramming after cell fusion between transplanted MSCs and recipient cardiac cells. We used a Cre/LoxP-based luciferase reporter system coupled to biophotonic imaging to detect fusion of transplanted human pluripotent stem cell-derived MSCs to cells of organs of living mice. Human MSCs, with transient expression of a viral fusogen, were delivered to the murine heart via a collagen patch. At 2 days and 1 week later, living mice were probed for bioluminescence indicative of cell fusion. Cell fusion was detected at the site of delivery (heart) and in distal tissues (i.e., stomach, small intestine, liver). Fusion was confirmed at the cellular scale via fluorescence in situ hybridization for human-specific and mouse-specific centromeres. Human cells in organs distal to the heart were typically located near the vasculature, suggesting MSCs and perhaps MSC fusion products have the ability to migrate via the circulatory system to distal organs and engraft with local cells. The present study reveals previously unknown migratory patterns of delivered human MSCs and associated fusion products in the healthy murine heart. The study also sets the stage for follow-on studies to determine the functional effects of cell fusion in a model of myocardial damage or disease. Mesenchymal stem cells (MSCs) are transplanted to the heart, cartilage, and other tissues to recover lost function or at least limit overactive immune responses. Analysis of tissues after MSC transplantation shows evidence of fusion between MSCs and the cells of the recipient. To date, the biologic implications of cell fusion remain unclear. A newly developed in vivo tracking system was used to identify MSC fusion products in living mice. The migratory patterns of fusion products were determined both in the target organ (i

  2. Multiple Convective Cell Identification and Tracking Algorithm for documenting time-height evolution of measured polarimetric radar and lightning properties

    Rosenfeld, D.; Hu, J.; Zhang, P.; Snyder, J.; Orville, R. E.; Ryzhkov, A.; Zrnic, D.; Williams, E.; Zhang, R.

    2017-12-01

    A methodology to track the evolution of the hydrometeors and electrification of convective cells is presented and applied to various convective clouds from warm showers to super-cells. The input radar data are obtained from the polarimetric NEXRAD weather radars, The information on cloud electrification is obtained from Lightning Mapping Arrays (LMA). The development time and height of the hydrometeors and electrification requires tracking the evolution and lifecycle of convective cells. A new methodology for Multi-Cell Identification and Tracking (MCIT) is presented in this study. This new algorithm is applied to time series of radar volume scans. A cell is defined as a local maximum in the Vertical Integrated Liquid (VIL), and the echo area is divided between cells using a watershed algorithm. The tracking of the cells between radar volume scans is done by identifying the two cells in consecutive radar scans that have maximum common VIL. The vertical profile of the polarimetric radar properties are used for constructing the time-height cross section of the cell properties around the peak reflectivity as a function of height. The LMA sources that occur within the cell area are integrated as a function of height as well for each time step, as determined by the radar volume scans. The result of the tracking can provide insights to the evolution of storms, hydrometer types, precipitation initiation and cloud electrification under different thermodynamic, aerosol and geographic conditions. The details of the MCIT algorithm, its products and their performance for different types of storm are described in this poster.

  3. Managing magnetic nanoparticle aggregation and cellular uptake: a precondition for efficient stem-cell differentiation and MRI tracking.

    Fayol, Delphine; Luciani, Nathalie; Lartigue, Lenaic; Gazeau, Florence; Wilhelm, Claire

    2013-02-01

    The labeling of stem cells with iron oxide nanoparticles is increasingly used to enable MRI cell tracking and magnetic cell manipulation, stimulating the fields of tissue engineering and cell therapy. However, the impact of magnetic labeling on stem-cell differentiation is still controversial. One compromising factor for successful differentiation may arise from early interactions of nanoparticles with cells during the labeling procedure. It is hypothesized that the lack of control over nanoparticle colloidal stability in biological media may lead to undesirable nanoparticle localization, overestimation of cellular uptake, misleading MRI cell tracking, and further impairment of differentiation. Herein a method is described for labeling mesenchymal stem cells (MSC), in which the physical state of citrate-coated nanoparticles (dispersed versus aggregated) can be kinetically tuned through electrostatic and magnetic triggers, as monitored by diffusion light scattering in the extracellular medium and by optical and electronic microscopy in cells. A set of statistical cell-by-cell measurements (flow cytometry, single-cell magnetophoresis, and high-resolution MRI cellular detection) is used to independently quantify the nanoparticle cell uptake and the effects of nanoparticle aggregation. Such aggregation confounds MRI cell detection as well as global iron quantification and has adverse effects on chondrogenetic differentiation. Magnetic labeling conditions with perfectly stable nanoparticles-suitable for obtaining differentiation-capable magnetic stem cells for use in cell therapy-are subsequently identified. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Making tracks

    Anon.

    1986-10-15

    In many modern tracking chambers, the sense wires, rather than being lined up uniformly, are grouped into clusters to facilitate the pattern recognition process. However, with higher energy machines providing collisions richer in secondary particles, event reconstruction becomes more complicated. A Caltech / Illinois / SLAC / Washington group developed an ingenious track finding and fitting approach for the Mark III detector used at the SPEAR electron-positron ring at SLAC (Stanford). This capitalizes on the detector's triggering, which uses programmable logic circuits operating in parallel, each 'knowing' the cell patterns for all tracks passing through a specific portion of the tracker (drift chamber)

  5. Tracking targeted bimodal nanovaccines: immune responses and routing in cells, tissue, and whole organism.

    Cruz, Luis J; Tacken, Paul J; Zeelenberg, Ingrid S; Srinivas, Mangala; Bonetto, Fernando; Weigelin, Bettina; Eich, Christina; de Vries, I Jolanda; Figdor, Carl G

    2014-12-01

    Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs), involved in the induction of immunity and currently exploited for antitumor immunotherapies. An optimized noninvasive imaging modality capable of determining and quantifying DC-targeted nanoparticle (NP) trajectories could provide valuable information regarding therapeutic vaccine outcome. Here, targeted poly(d,l-lactide-co-glycolide) nanoparticles (PLGA NPs) recognizing DC receptors were equipped with superparamagnetic iron oxide particles (SPIO) or gold nanoparticles with fluorescently labeled antigen. The fluorescent label allowed for rapid analysis and quantification of DC-specific uptake of targeted PLGA NPs in comparison to uptake by other cells. Transmission electron microscopy (TEM) showed that a fraction of the encapsulated antigen reached the lysosomal compartment of DCs, where SPIO and gold were already partially released. However, part of the PLGA NPs localized within the cytoplasm, as confirmed by confocal microscopy. DCs targeted with NPs carrying SPIO or fluorescent antigen were detected within lymph nodes as early as 1 h after injection by magnetic resonance imaging (MRI). Despite the fact that targeting did not markedly affect PLGA NP biodistribution on organism and tissue level, it increased delivery of NPs to DCs residing in peripheral lymph nodes and resulted in enhanced T cell proliferation. In conclusion, two imaging agents within a single carrier allows tracking of targeted PLGA NPs at the subcellular, cellular, and organismal levels, thereby facilitating the rational design of in vivo targeted vaccination strategies.

  6. Magnetic poly(lactide-co-glycolide) (PLGA) and cellulose particles for MRI-based cell tracking

    Nkansah, Michael K.; Thakral, Durga; Shapiro, Erik M.

    2010-01-01

    Biodegradable, superparamagnetic micro- and nanoparticles of poly(lactide-co-glycolide) (PLGA) and cellulose were designed, fabricated and characterized for magnetic cell labeling. Monodisperse nanocrystals of magnetite were incorporated into micro- and nanoparticles of PLGA and cellulose with high efficiency using an oil-in-water single emulsion technique. Superparamagnetic cores had high magnetization (72.1 emu/g). The resulting polymeric particles had smooth surface morphology and high magnetite content (43.3 wt% for PLGA and 69.6 wt% for cellulose). While PLGA and cellulose nanoparticles displayed highest r2* values per millimole of iron (399 s-1mM-1 for cellulose and 505 s-1mM-1 for PLGA), micron-sized PLGA particles had a much higher r2* per particle than either. After incubation for a month in citrate buffer (pH 5.5), magnetic PLGA particles lost close to 50% of their initial r2* molar relaxivity, while magnetic cellulose particles remained intact, preserving over 85% of their initial r2* molar relaxivity. Lastly, mesenchymal stem cells and human breast adenocarcinoma cells were magnetically labeled using these particles with no detectable cytotoxicity. These particles are ideally suited for non-invasive cell tracking in vivo via MRI and due to their vastly different degradation properties, offer unique potential for dedicated use for either short (PLGA-based particles) or long term (cellulose-based particles) experiments. PMID:21404328

  7. Flow Profile Study using miniature Laser-Doppler velocimetry

    Booij, W.E.; Booij, W.E.; de Jongh, A.; de Mul, F.F.M.

    1995-01-01

    We present a physics experiment, in which laser - Doppler velocimetry is used to make first - year university physics students realize that the idealized solutions offered by standard text books seldom are applicable without corrections, which often are numerical. This is demonstrated by carefully

  8. Broadband phase difference method for ultrasonic velocimetry in molten glass

    Kikura, Hiroshige; Ihara, Tomonori

    2016-01-01

    This study aims to develop ultrasonic Doppler velocimetry in molten glass. Realization of such a technique has two difficulties: ultrasonic transmission into molten salt and Doppler signal processing. Buffer rod technique was developed in our research to transmit ultrasound into high temperature molten glass. This article discusses newly developed signal processing technique named broadband phase difference method. (J.P.N.)

  9. Reusable holographic velocimetry system based on polarization multiplexing in Bacteriorhodopsin

    Koek, W.D.; Chan, V.S.S.; Ooms, T.A.; Bhattacharya, N.; Westerweel, J.; Braat, J.J.M.

    2005-01-01

    We present a novel holographic particle image velocimetry (HPIV) system using a reversible holographic material as the recording medium. In HPIV the three-dimensional flow field throughout a volume is detected by adding small tracer particles to a normally transparent medium. By recording the

  10. Quantum measurement and orientation tracking of fluorescent nanodiamonds inside living cells

    McGuinness, L. P.; Yan, Y.; Stacey, A.; Simpson, D. A.; Hall, L. T.; MacLaurin, D.; Prawer, S.; Mulvaney, P.; Wrachtrup, J.; Caruso, F.; Scholten, R. E.; Hollenberg, L. C. L.

    2011-06-01

    Fluorescent particles are routinely used to probe biological processes. The quantum properties of single spins within fluorescent particles have been explored in the field of nanoscale magnetometry, but not yet in biological environments. Here, we demonstrate optically detected magnetic resonance of individual fluorescent nanodiamond nitrogen-vacancy centres inside living human HeLa cells, and measure their location, orientation, spin levels and spin coherence times with nanoscale precision. Quantum coherence was measured through Rabi and spin-echo sequences over long (>10 h) periods, and orientation was tracked with effective 1° angular precision over acquisition times of 89 ms. The quantum spin levels served as fingerprints, allowing individual centres with identical fluorescence to be identified and tracked simultaneously. Furthermore, monitoring decoherence rates in response to changes in the local environment may provide new information about intracellular processes. The experiments reported here demonstrate the viability of controlled single spin probes for nanomagnetometry in biological systems, opening up a host of new possibilities for quantum-based imaging in the life sciences.

  11. Time-Lapse Monitoring of DNA Damage Colocalized With Particle Tracks in Single Living Cells

    McFadden, Conor H. [Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Hallacy, Timothy M. [Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Department of Physics and Astronomy, Rice University, Houston, Texas (United States); Flint, David B. [Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Graduate School of Biomedical Sciences, The University of Texas, Houston, Texas (United States); Granville, Dal A. [Department of Medical Physics, The Ottawa Hospital Cancer Centre, Ottawa, Ontario (Canada); Asaithamby, Aroumougame [Division of Molecular Radiation Biology, Department of Radiation Oncology, University of Texas Southwestern Medical Centre, Dallas, Texas (United States); Sahoo, Narayan [Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Akselrod, Mark S. [Crystal Growth Division, Landauer, Inc, Stillwater, Oklahoma (United States); Sawakuchi, Gabriel O., E-mail: gsawakuchi@mdanderson.org [Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, Texas (United States); Graduate School of Biomedical Sciences, The University of Texas, Houston, Texas (United States)

    2016-09-01

    Purpose: Understanding the DNA damage and repair induced by hadron therapy (HT) beams is crucial for developing novel strategies to maximize the use of HT beams to treat cancer patients. However, spatiotemporal studies of DNA damage and repair for beam energies relevant to HT have been challenging. We report a technique that enables spatiotemporal measurement of radiation-induced damage in live cells and colocalization of this damage with charged particle tracks over a broad range of clinically relevant beam energies. The technique uses novel fluorescence nuclear track detectors with fluorescence confocal laser scanning microscopy in the beam line to visualize particle track traversals within the subcellular compartments of live cells within seconds after injury. Methods and Materials: We designed and built a portable fluorescence confocal laser scanning microscope for use in the beam path, coated fluorescence nuclear track detectors with fluorescent-tagged live cells (HT1080 expressing enhanced green fluorescent protein tagged to XRCC1, a single-strand break repair protein), placed the entire assembly into a proton therapy beam line, and irradiated the cells with a fluence of ∼1 × 10{sup 6} protons/cm{sup 2}. Results: We successfully obtained confocal images of proton tracks and foci of DNA single-strand breaks immediately after irradiation. Conclusions: This technique represents an innovative method for analyzing biological responses in any HT beam line at energies and dose rates relevant to therapy. It allows precise determination of the number of tracks traversing a subcellular compartment and monitoring the cellular damage therein, and has the potential to measure the linear energy transfer of each track from therapeutic beams.

  12. Tracking shocked dust: State estimation for a complex plasma during a shock wave

    Oxtoby, Neil P.; Ralph, Jason F.; Durniak, Celine; Samsonov, Dmitry

    2012-01-01

    We consider a two-dimensional complex (dusty) plasma crystal excited by an electrostatically-induced shock wave. Dust particle kinematics in such a system are usually determined using particle tracking velocimetry. In this work we present a particle tracking algorithm which determines the dust particle kinematics with significantly higher accuracy than particle tracking velocimetry. The algorithm uses multiple extended Kalman filters to estimate the particle states and an interacting multiple model to assign probabilities to the different filters. This enables the determination of relevant physical properties of the dust, such as kinetic energy and kinetic temperature, with high precision. We use a Hugoniot shock-jump relation to calculate a pressure-volume diagram from the shocked dust kinematics. Calculation of the full pressure-volume diagram was possible with our tracking algorithm, but not with particle tracking velocimetry.

  13. In vivo tracking of neuronal-like cells by magnetic resonance in rabbit models of spinal cord injury

    Zhang, Ruiping; Zhang, Kun; Li, Jianding; Liu, Qiang; Xie, Jun

    2013-01-01

    In vitro experiments have demonstrated that neuronal-like cells derived from bone marrow mesenchymal stem cells can survive, migrate, integrate and help to restore the function and behaviors of spinal cord injury models, and that they may serve as a suitable approach to treating spinal cord injury. However, it is very difficult to track transplanted cells in vivo. In this study, we injected superparamagnetic iron oxide-labeled neuronal-like cells into the subarachnoid space in a rabbit model of spinal cord injury. At 7 days after cell transplantation, a small number of dot-shaped low signal intensity shadows were observed in the spinal cord injury region, and at 14 days, the number of these shadows increased on T2-weighted imaging. Perl's Prussian blue staining detected dot-shaped low signal intensity shadows in the spinal cord injury region, indicative of superparamagnetic iron oxide nanoparticle-labeled cells. These findings suggest that transplanted neuronal-like cells derived from bone marrow mesenchymal stem cells can migrate to the spinal cord injury region and can be tracked by magnetic resonance in vivo. Magnetic resonance imaging represents an efficient noninvasive technique for visually tracking transplanted cells in vivo. PMID:25206659

  14. Developing new optical imaging techniques for single particle and molecule tracking in live cells

    Sun, Wei [Iowa State Univ., Ames, IA (United States)

    2010-01-01

    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. 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

  15. Nanohybrids with Magnetic and Persistent Luminescence Properties for Cell Labeling, Tracking, In Vivo Real-Time Imaging, and Magnetic Vectorization.

    Teston, Eliott; Maldiney, Thomas; Marangon, Iris; Volatron, Jeanne; Lalatonne, Yoann; Motte, Laurence; Boisson-Vidal, Catherine; Autret, Gwennhael; Clément, Olivier; Scherman, Daniel; Gazeau, Florence; Richard, Cyrille

    2018-04-01

    Once injected into a living organism, cells diffuse or migrate around the initial injection point and become impossible to be visualized and tracked in vivo. The present work concerns the development of a new technique for therapeutic cell labeling and subsequent in vivo visualization and magnetic retention. It is hypothesized and subsequently demonstrated that nanohybrids made of persistent luminescence nanoparticles and ultrasmall superparamagnetic iron oxide nanoparticles incorporated into a silica matrix can be used as an effective nanoplatform to label therapeutic cells in a nontoxic way in order to dynamically track them in real-time in vitro and in living mice. As a proof-of-concept, it is shown that once injected, these labeled cells can be visualized and attracted in vivo using a magnet. This first step suggests that these nanohybrids represent efficient multifunctional nanoprobes for further imaging guided cell therapies development. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Tracking by flow cytometry antigen-specific follicular helper T cells in wild-type animals after protein vaccination.

    Chakarov, Svetoslav; Fazilleau, Nicolas

    2015-01-01

    Flow cytometry is a valuable technology used in immunology to characterize and enumerate the different cell subpopulations specific for a nonself-antigen in the context of an ongoing immune response. Among them, follicular helper T cells are the cognate regulators of B cells in secondary lymphoid tissues. Thus, tracking them is of high interest especially in the context of protein vaccination. For this purpose, transgenic antigen-receptor mouse models have been largely used. It is now clear that transgenic models are not always the best means to study the dynamics of the immune response since they can modify the response. In this chapter, we describe how to track endogenous antigen-specific follicular helper T cells by flow cytometry after protein vaccination in nonmodified wild-type animals, which ultimately provides a comprehensive way to enumerate, characterize, and isolate these particular cells in vivo.

  17. Visualization and correction of automated segmentation, tracking and lineaging from 5-D stem cell image sequences.

    Wait, Eric; Winter, Mark; Bjornsson, Chris; Kokovay, Erzsebet; Wang, Yue; Goderie, Susan; Temple, Sally; Cohen, Andrew R

    2014-10-03

    Neural stem cells are motile and proliferative cells that undergo mitosis, dividing to produce daughter cells and ultimately generating differentiated neurons and glia. Understanding the mechanisms controlling neural stem cell proliferation and differentiation will play a key role in the emerging fields of regenerative medicine and cancer therapeutics. Stem cell studies in vitro from 2-D image data are well established. Visualizing and analyzing large three dimensional images of intact tissue is a challenging task. It becomes more difficult as the dimensionality of the image data increases to include time and additional fluorescence channels. There is a pressing need for 5-D image analysis and visualization tools to study cellular dynamics in the intact niche and to quantify the role that environmental factors play in determining cell fate. We present an application that integrates visualization and quantitative analysis of 5-D (x,y,z,t,channel) and large montage confocal fluorescence microscopy images. The image sequences show stem cells together with blood vessels, enabling quantification of the dynamic behaviors of stem cells in relation to their vascular niche, with applications in developmental and cancer biology. Our application automatically segments, tracks, and lineages the image sequence data and then allows the user to view and edit the results of automated algorithms in a stereoscopic 3-D window while simultaneously viewing the stem cell lineage tree in a 2-D window. Using the GPU to store and render the image sequence data enables a hybrid computational approach. An inference-based approach utilizing user-provided edits to automatically correct related mistakes executes interactively on the system CPU while the GPU handles 3-D visualization tasks. By exploiting commodity computer gaming hardware, we have developed an application that can be run in the laboratory to facilitate rapid iteration through biological experiments. We combine unsupervised image

  18. Moving epithelia: Tracking the fate of mammalian limbal epithelial stem cells.

    Di Girolamo, Nick

    2015-09-01

    Lineage tracing allows the destiny of a stem cell (SC) and its progeny to be followed through time. In order to track their long-term fate, SC must be permanently marked to discern their distribution, division, displacement and differentiation. This information is essential for unravelling the mysteries that govern their replenishing activity while they remain anchored within their niche microenvironment. Modern-day lineage tracing uses inducible genetic recombination to illuminate cells within embryonic, newborn and adult tissues, and the advent of powerful high-resolution microscopy has enabled the behaviour of labelled cells to be monitored in real-time in a living organism. The simple structural organization of the mammalian cornea, including its accessibility and transparency, renders it the ideal tissue to study SC fate using lineage tracing assisted by non-invasive intravital microscopy. Despite more than a century of research devoted to understanding how this tissue is maintained and repaired, many limitations and controversies continue to plague the field, including uncertainties about the specificity of current SC markers, the number of SC within the cornea, their mode of division, their location, and importantly the signals that dictate cell migration. This communication will highlight historical discoveries as well as recent developments in the corneal SC field; more specifically how the progeny of these cells are mobilised to replenish this dynamic tissue during steady-state, disease and transplantation. Also discussed is how insights gleaned from animal studies can be used to advance our knowledge of the fundamental mechanisms that govern modelling and remodelling of the human cornea in health and disease. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. SPECT Imaging for in vivo tracking of NIS containing stem cells

    Lee, Zhenghong

    2013-04-02

    The proposed study contains two groups of imaging experiments: 1) human mesenchymal stem cells supporting in vivo survival of unrelated donor hematopoietic stem cells; 2) gene transduction and selection of mutant MGMT genes on human hematopoietic stem cells conferring resistance to BC+BCNU. There is increasing evidence that adult human tissues harbor stem and progenitor cells that can be used for therapeutic purposes. We had focused on the Mesenchymal Stem Cells (MSCs) found in human bone marrow and investigated these cells in the context of autologous and allogeneic hematopoietic stem cell transplantation to a) facilitate rapid hematopoietic engraftment in cancer patients receiving high dose chemotherapy and b) to modulate the graft-versus-host disease (GVHD). We have demonstrated that culture-expanded autologous and allogeneic MSCs can be safely infused into humans and the preliminary results showed that MSCs facilitate hematopoietic engraftment and reduce GVHD. On the other hand, studies of gene transfer with drug resistant selection suggest major perturbations to the process of hematopoietic reconstitution and the confounding issue of organ toxicity and recovery that takes place in the host. We have found that limiting numbers of hematopoietic stem cells transduced with MGMT repopulate the bone marrow of primary and secondary recipient mice. We are also particularly interested in the dynamics of engraftment and selection in regions of bones, liver, spleen and lung, where we have previously seen marked evidence of engraftment. All the measurements have required animal sacrifice and single point determinations of engraftment in individual and cohorts of mice. Heretofore it has not been possible to study the dynamics of engraftment and enrichment. In the upcoming application, we propose to develop an imaging method to track intravenously infused stem cells in vivo at preset time points to understand their homing and proliferation. Specifically, we propose to use

  20. Convergence of lateral dynamic measurements in the plasma membrane of live cells from single particle tracking and STED-FCS

    Lagerholm, B. Christoffer; Andrade, Débora M.; Clausen, Mathias P.

    2017-01-01

    Fluorescence correlation spectroscopy (FCS) in combination with the super-resolution imaging method STED (STED-FCS), and single-particle tracking (SPT) are able to directly probe the lateral dynamics of lipids and proteins in the plasma membrane of live cells at spatial scales much below the diff...

  1. "Sickle Cell Anemia: Tracking down a Mutation": An Interactive Learning Laboratory That Communicates Basic Principles of Genetics and Cellular Biology

    Jarrett, Kevin; Williams, Mary; Horn, Spencer; Radford, David; Wyss, J. Michael

    2016-01-01

    "Sickle cell anemia: tracking down a mutation" is a full-day, inquiry-based, biology experience for high school students enrolled in genetics or advanced biology courses. In the experience, students use restriction endonuclease digestion, cellulose acetate gel electrophoresis, and microscopy to discover which of three putative patients…

  2. The effect of track structure on the induction of chromosomal aberrations in murine cells

    Durante, M.; Cella, L.; Furusawa, Y.; George, K.; Gialanella, G.; Grossi, G.; Pugliese, M.; Saito, M.; Yang, T. C.

    1998-01-01

    PURPOSE: To measure chromosome aberrations in C3H 10T1/2 mouse fibroblasts using FISH painting at the first mitosis following exposure to 30 keV/microm hydrogen or neon ions. MATERIALS AND METHODS: Cells in plateau-phase were irradiated with 0.86 MeV protons at the TTT-3 Tandem accelerator in Naples (Italy), or with 400 MeV/n Ne ions at the HIMAC accelerator in Chiba (Japan). Colcemid-blocked cells were harvested at the first mitosis following exposure, and chromosome spreads were hybridized in situ with a fluorescein-labelled composite mouse DNA probe specific for chromosomes 2 and 8. RESULTS: Protons were more efficient than neon ions at the same LET in the induction of chromosome interchanges and breaks. Yields of complex exchanges were similar for both particles at the same dose, but protons produced mostly insertions, while with Ne exposure non-reciprocal exchanges were the most frequent complex-type exchange. CONCLUSIONS: Charged particles with the same LET produce different yields of chromosome aberrations, and some observed differences can be explained based on the available track-structure models.

  3. Theranostic Niosomes for Efficient siRNA/microRNA Delivery and Activatable Near-Infrared Fluorescent Tracking of Stem Cells

    Yang, Chuanxu; Shan, Gao; Song, Ping

    2018-01-01

    RNA interference (RNAi) mediated gene regulation in stem cells offers great potential in regenerative medicine. In this study, we developed a theranostic platform for efficient delivery of small RNAs (siRNA/miRNA) to human mesenchymal stem cells (hMSCs) to promote differentiation, and meanwhile...... OFF/ON activatable fluorescence upon cellular internalization, resulting in efficient NIR labeling and the capability to dynamically monitor stem cells in mice. In addition, iSPN/siRNA achieved simultaneous long-term cell tracking and in vivo gene silencing after implantation in mice. These results...

  4. Exploring Transduction Mechanisms of Protein Transduction Domains (PTDs in Living Cells Utilizing Single-Quantum Dot Tracking (SQT Technology

    Yasuhiro Suzuki

    2012-01-01

    Full Text Available Specific protein domains known as protein transduction domains (PTDs can permeate cell membranes and deliver proteins or bioactive materials into living cells. Various approaches have been applied for improving their transduction efficacy. It is, therefore, crucial to clarify the entry mechanisms and to identify the rate-limiting steps. Because of technical limitations for imaging PTD behavior on cells with conventional fluorescent-dyes, how PTDs enter the cells has been a topic of much debate. Utilizing quantum dots (QDs, we recently tracked the behavior of PTD that was derived from HIV-1 Tat (TatP in living cells at the single-molecule level with 7-nm special precision. In this review article, we initially summarize the controversy on TatP entry mechanisms; thereafter, we will focus on our recent findings on single-TatP-QD tracking (SQT, to identify the major sequential steps of intracellular delivery in living cells and to discuss how SQT can easily provide direct information on TatP entry mechanisms. As a primer for SQT study, we also discuss the latest findings on single particle tracking of various molecules on the plasma membrane. Finally, we discuss the problems of QDs and the challenges for the future in utilizing currently available QD probes for SQT. In conclusion, direct identification of the rate-limiting steps of PTD entry with SQT should dramatically improve the methods for enhancing transduction efficiency.

  5. Single charged-particle damage to living cells: a new method based on track-etch detectors

    Durante, M.; Grossi, G.F.; Pugliese, M.; Manti, L.; Nappo, M.; Gialanella, G.

    1994-01-01

    Biological effects of ionizing radiation are usually expressed as a function of the absorbed dose. Low doses of high-LET radiation correspond to one or few particle traversals through the cell. In order to study the biological effectiveness of single charged particles, we have developed a new method based on solid state nuclear track detectors. Cells are seeded on mylar and a LR-115 film is stuck below the mylar base. After irradiation, the LR-115 film is etched and cells observed at a phase contrast microscope connected to a video camera and an image analyzer. In this way, it is possible to measure the number of traversals through the cell nucleus or cytoplasm. Coordinates of each cell on the microscope bench are saved. After incubation for about one week, cells are fixed and stained and the colonies observed at the microscope. The fate of each irradiated cell is therefore correlated to the number of traversals. We have tested this method with two different rodent embryo fibroblast cell lines, C3H 10T1/2 and V79, exposed to 3.2 MeV accelerated α-particles (LET =124 keV/μm). The studied endpoint was cell killing. Preliminary biological results suggest that few α-particle tracks in V79 hamster cells are sufficient to reduce surviving fraction. ((orig.))

  6. Ultrastructural characterization of mesenchymal stromal cells labeled with ultrasmall superparamagnetic iron-oxide nanoparticles for clinical tracking studies

    Hansen, Louise; Hansen, Alastair B; Mathiasen, Anders B

    2014-01-01

    INTRODUCTION: To evaluate survival and engraftment of mesenchymal stromal cells (MSCs) in vivo, it is necessary to track implanted cells non-invasively with a method, which does not influence cellular ultrastructure and functional characteristics. Iron-oxide particles have been applied for cell...... sequence of trans-activator of transcription (TAT) (IODEX-TAT) and evaluate the effect of labeling on ultrastructure, viability, phenotype and proliferative capacity of the cells. MATERIALS AND METHODS: MSCs were labeled with 5 and 10 μg IODEX-TAT/10(5) cells for 2, 6 and 21 hours. IODEX-TAT uptake...... and cellular ultrastructure were determined by electron microscopy. Cell viability was determined by propidium iodide staining and cell proliferation capacity by 5-bromo-2-deoxyuridine (BrdU) incorporation. Maintenance of stem cell surface markers was determined by flow cytometry. Results. IODEX-TAT labeling...

  7. Iced airfoil separation bubble measurements by particle image velocimetry

    Jacobs, Jason J.

    Not long after the birth of aviation, pilots began to recognize the dangers posed by aircraft icing. Since that time, research has improved the awareness of this problem and the scientific understanding of the associated aerodynamic impacts, however, few studies have involved detailed, quantitative, flowfield measurements. For this reason, the current investigation was conducted in which high spatial-resolution flowfield measurements were acquired of a NACA 0012 airfoil with two- and three-dimensional, simulated, leading-edge, horn-ice accretions utilizing particle image velocimetry (PIV). These measurements complemented existing iced airfoil performance measurements, revealed previously unknown details regarding the structure and behavior of these flowfields, and could potentially facilitate the development and improvement of computational schemes used to predict largely separated flows, including that of an iced airfoil near stall. Previous iced airfoil investigations have demonstrated somewhat reduced aerodynamic penalties resulting from a three-dimensional ice simulation, compared to those of a two-dimensional ice simulation of a representative cross section. Correspondingly, the current measurements revealed accelerated transition of the separated shear layer emanating from a three-dimensional ice simulation and therefore enhanced pressure recovery and reduced mean separation bubble length, each relative to the flowfield of a representative two-dimensional ice simulation. These effects appeared to result from the quasi-steady distribution of discrete, streamwise vortices which aided the turbulent entrainment of fluid from the recirculation region of the three-dimensional ice simulation separation bubble flowfield. These vortices were generated by a streamwise-vortex instability excited by roughness along the three-dimensional ice simulation and produced spanwise-cell structures throughout this flowfield, as well as significant spanwise variation in peak

  8. Laser speckle velocimetry applied to Rayleigh-Benard convection

    Arroyo, M.P.; Yonte, T.; Quintanilla, M.; Saviron, J.M.

    1986-01-01

    An application of speckle velocimetry technique to Rayleigh-Benard convection is presented. A 5-mW He-Ne laser allows precise determination of the two-dimensional velocity flow field, up to several mm/sec. The digital techniques used to analyze automatically the multiexposed photographs and to generate velocity and vorticity fields are described. The obtained results are in good agreement with previously reported data. The ability of the technique to cover other experimental conditions is discussed. 14 references

  9. Large scale particle image velocimetry with helium filled soap bubbles

    Bosbach, Johannes; Kuehn, Matthias; Wagner, Claus [German Aerospace Center (DLR), Institute of Aerodynamics and Flow Technology, Goettingen (Germany)

    2009-03-15

    The application of particle image velocimetry (PIV) to measurement of flows on large scales is a challenging necessity especially for the investigation of convective air flows. Combining helium filled soap bubbles as tracer particles with high power quality switched solid state lasers as light sources allows conducting PIV on scales of the order of several square meters. The technique was applied to mixed convection in a full scale double aisle aircraft cabin mock-up for validation of computational fluid dynamics simulations. (orig.)

  10. Large scale particle image velocimetry with helium filled soap bubbles

    Bosbach, Johannes; Kühn, Matthias; Wagner, Claus

    2009-03-01

    The application of Particle Image Velocimetry (PIV) to measurement of flows on large scales is a challenging necessity especially for the investigation of convective air flows. Combining helium filled soap bubbles as tracer particles with high power quality switched solid state lasers as light sources allows conducting PIV on scales of the order of several square meters. The technique was applied to mixed convection in a full scale double aisle aircraft cabin mock-up for validation of Computational Fluid Dynamics simulations.

  11. Particle Image Velocimetry Applications of Fluorescent Dye-Doped Particles

    Petrosky, Brian Joseph

    2015-01-01

    Laser flare can often be a major issue in particle image velocimetry (PIV) involving solid boundaries in a flow or a gas-liquid interface. The use of fluorescent light from dye-doped particles has been demonstrated in water applications, but reproducing the technique in an airflow is more difficult due to particle size constraints and safety concerns. The following thesis is formatted in a hybrid manuscript style, including a full paper presenting the applications of fluorescent Kiton R...

  12. Particle Image Velocimetry Applications Using Fluorescent Dye-Doped Particles

    Petrosky, Brian J.; Maisto, Pietro; Lowe, K. Todd; Andre, Matthieu A.; Bardet, Philippe M.; Tiemsin, Patsy I.; Wohl, Christopher J.; Danehy, Paul M.

    2015-01-01

    Polystyrene latex sphere particles are widely used to seed flows for velocimetry techniques such as Particle Image Velocimetry (PIV) and Laser Doppler Velocimetry (LDV). These particles may be doped with fluorescent dyes such that signals spectrally shifted from the incident laser wavelength may be detected via Laser Induced Fluorescence (LIF). An attractive application of the LIF signal is achieving velocimetry in the presence of strong interference from laser scatter, opening up new research possibilities very near solid surfaces or at liquid/gas interfaces. Additionally, LIF signals can be used to tag different fluid streams to study mixing. While fluorescence-based PIV has been performed by many researchers for particles dispersed in water flows, the current work is among the first in applying the technique to micron-scale particles dispersed in a gas. A key requirement for such an application is addressing potential health hazards from fluorescent dyes; successful doping of Kiton Red 620 (KR620) has enabled the use of this relatively safe dye for fluorescence PIV for the first time. In this paper, basic applications proving the concept of PIV using the LIF signal from KR620-doped particles are exhibited for a free jet and a twophase flow apparatus. Results indicate that while the fluorescence PIV techniques are roughly 2 orders of magnitude weaker than Mie scattering, they provide a viable method for obtaining data in flow regions previously inaccessible via standard PIV. These techniques have the potential to also complement Mie scattering signals, for example in multi-stream and/or multi-phase experiments.

  13. Lateral motion and bending of microtubules studied with a new single-filament tracking routine in living cells.

    Pallavicini, Carla; Levi, Valeria; Wetzler, Diana E; Angiolini, Juan F; Benseñor, Lorena; Despósito, Marcelo A; Bruno, Luciana

    2014-06-17

    The cytoskeleton is involved in numerous cellular processes such as migration, division, and contraction and provides the tracks for transport driven by molecular motors. Therefore, it is very important to quantify the mechanical behavior of the cytoskeletal filaments to get a better insight into cell mechanics and organization. It has been demonstrated that relevant mechanical properties of microtubules can be extracted from the analysis of their motion and shape fluctuations. However, tracking individual filaments in living cells is extremely complex due, for example, to the high and heterogeneous background. We introduce a believed new tracking algorithm that allows recovering the coordinates of fluorescent microtubules with ∼9 nm precision in in vitro conditions. To illustrate potential applications of this algorithm, we studied the curvature distributions of fluorescent microtubules in living cells. By performing a Fourier analysis of the microtubule shapes, we found that the curvatures followed a thermal-like distribution as previously reported with an effective persistence length of ∼20 μm, a value significantly smaller than that measured in vitro. We also verified that the microtubule-associated protein XTP or the depolymerization of the actin network do not affect this value; however, the disruption of intermediate filaments decreased the persistence length. Also, we recovered trajectories of microtubule segments in actin or intermediate filament-depleted cells, and observed a significant increase of their motion with respect to untreated cells showing that these filaments contribute to the overall organization of the microtubule network. Moreover, the analysis of trajectories of microtubule segments in untreated cells showed that these filaments presented a slower but more directional motion in the cortex with respect to the perinuclear region, and suggests that the tracking routine would allow mapping the microtubule dynamical organization in cells

  14. Iodine Tagging Velocimetry in a Mach 10 Wake

    Balla, Robert Jeffrey

    2013-01-01

    A variation on molecular tagging velocimetry (MTV) [1] designated iodine tagging velocimetry (ITV) is demonstrated. Molecular iodine is tagged by two-photon absorption using an Argon Fluoride (ArF) excimer laser. A single camera measures fluid displacement using atomic iodine emission at 206 nm. Two examples ofMTVfor cold-flowmeasurements areN2OMTV [2] and Femtosecond Laser Electronic Excitation Tagging [3]. These, like most MTV methods, are designed for atmospheric pressure applications. Neither can be implemented at the low pressures (0.1- 1 Torr) in typical hypersonic wakes. Of all the single-laser/singlecamera MTV approaches, only Nitric-Oxide Planar Laser Induced Fluorescence-based MTV [4] has been successfully demonstrated in a Mach 10 wake. Oxygen quenching limits transit times to 500 ns and accuracy to typically 30%. The present note describes the photophysics of the ITV method. Off-body velocimetry along a line is demonstrated in the aerothermodynamically important and experimentally challenging region of a hypersonic low-pressure near-wake in a Mach 10 air wind tunnel. Transit times up to 10 µs are demonstrated with conservative errors of 10%.

  15. A tracer liquid image velocimetry for multi-layer radial flow in bioreactors.

    Gao, Yu-Bao; Liang, Jiu-Xing; Luo, Yu-Xi; Yan, Jia

    2015-02-13

    This paper presents a Tracer Liquid Image Velocimetry (TLIV) for multi-layer radial flow in bioreactors used for cells cultivation of tissue engineering. The goal of this approach is to use simple devices to get good measuring precision, specialized for the case in which the uniform level of fluid shear stress was required while fluid velocity varied smoothly. Compared to the widely used Particles Image Velocimetry (PIV), this method adopted a bit of liquid as tracer, without the need of laser source. Sub-pixel positioning algorithm was used to overcome the adverse effects of the tracer liquid deformation. In addition, a neighborhood smoothing algorithm was used to restrict the measurement perturbation caused by diffusion. Experiments were carried out in a parallel plates flow chamber. And mathematical models of the flow chamber and Computational Fluid Dynamics (CFD) simulation were separately employed to validate the measurement precision of TLIV. The mean relative error between the simulated and measured data can be less than 2%, while in similar validations using PIV, the error was around 8.8%. TLIV avoided the contradiction between the particles' visibility and following performance with tested fluid, which is difficult to overcome in PIV. And TLIV is easier to popularize for its simple experimental condition and low cost.

  16. A Maximum Power Point Tracking Control Method of a Photovoltaic Power Generator with Consideration of Dynamic Characteristics of Solar Cells

    Watanabe, Takashi; Yoshida, Toshiya; Ohniwa, Katsumi

    This paper discusses a new control strategy for photovoltaic power generation systems with consideration of dynamic characteristics of the photovoltaic cells. The controller estimates internal currents of an equivalent circuit for the cells. This estimated, or the virtual current and the actual voltage of the cells are fed to a conventional Maximum-Power-Point-Tracking (MPPT) controller. Consequently, this MPPT controller still tracks the optimum point even though it is so designed that the seeking speed of the operating point is extremely high. This system may suit for applications, which are installed in rapidly changeable insolation and temperature-conditions e.g. automobiles, trains, and airplanes. The proposed method is verified by experiment with a combination of this estimating function and the modified Boehringer's MPPT algorithm.

  17. Assessing the efficacy of nano- and micro-sized magnetic particles as contrast agents for MRI cell tracking.

    Arthur Taylor

    Full Text Available Iron-oxide based contrast agents play an important role in magnetic resonance imaging (MRI of labelled cells in vivo. Currently, a wide range of such contrast agents is available with sizes varying from several nanometers up to a few micrometers and consisting of single or multiple magnetic cores. Here, we evaluate the effectiveness of these different particles for labelling and imaging stem cells, using a mouse mesenchymal stem cell line to investigate intracellular uptake, retention and processing of nano- and microsized contrast agents. The effect of intracellular confinement on transverse relaxivity was measured by MRI at 7 T and in compliance with the principles of the '3Rs', the suitability of the contrast agents for MR-based cell tracking in vivo was tested using a chick embryo model. We show that for all particles tested, relaxivity was markedly reduced following cellular internalisation, indicating that contrast agent relaxivity in colloidal suspension does not accurately predict performance in MR-based cell tracking studies. Using a bimodal imaging approach comprising fluorescence and MRI, we demonstrate that labelled MSC remain viable following in vivo transplantation and can be tracked effectively using MRI. Importantly, our data suggest that larger particles might confer advantages for longer-term imaging.

  18. Co-visualization of DNA damage and ion traversals in live mammalian cells using a fluorescent nuclear track detector

    Kodaira, Satoshi; Konishi, Teruaki; Kobayashi, Alisa

    2015-01-01

    The geometric locations of ion traversals in mammalian cells constitute important information in the study of heavy ion-induced biological effect. Single ion traversal through a cellular nucleus produces complex and massive DNA damage at a nanometer level, leading to cell inactivation, mutations and transformation. We present a novel approach that uses a fluorescent nuclear track detector (FNTD) for the simultaneous detection of the geometrical images of ion traversals and DNA damage in single cells using confocal microscopy. HT1080 or HT1080–53BP1-GFP cells were cultured on the surface of a FNTD and exposed to 5.1-MeV/n neon ions. The positions of the ion traversals were obtained as fluorescent images of a FNTD. Localized DNA damage in cells was identified as fluorescent spots of γ-H2AX or 53BP1-GFP. These track images and images of damaged DNA were obtained in a short time using a confocal laser scanning microscope. The geometrical distribution of DNA damage indicated by fluorescent γ-H2AX spots in fixed cells or fluorescent 53BP1-GFP spots in living cells was found to correlate well with the distribution of the ion traversals. This method will be useful for evaluating the number of ion hits on individual cells, not only for micro-beam but also for random-beam experiments. (author)

  19. Particle displacement tracking for PIV

    Wernet, Mark P.

    1990-01-01

    A new Particle Imaging Velocimetry (PIV) data acquisition and analysis system, which is an order of magnitude faster than any previously proposed system has been constructed and tested. The new Particle Displacement Tracing (PDT) system is an all electronic technique employing a video camera and a large memory buffer frame-grabber board. Using a simple encoding scheme, a time sequence of single exposure images are time coded into a single image and then processed to track particle displacements and determine velocity vectors. Application of the PDT technique to a counter-rotating vortex flow produced over 1100 velocity vectors in 110 seconds when processed on an 80386 PC.

  20. WE-H-BRA-08: A Monte Carlo Cell Nucleus Model for Assessing Cell Survival Probability Based On Particle Track Structure Analysis

    Lee, B [Northwestern Memorial Hospital, Chicago, IL (United States); Georgia Institute of Technology, Atlanta, GA (Georgia); Wang, C [Georgia Institute of Technology, Atlanta, GA (Georgia)

    2016-06-15

    Purpose: To correlate the damage produced by particles of different types and qualities to cell survival on the basis of nanodosimetric analysis and advanced DNA structures in the cell nucleus. Methods: A Monte Carlo code was developed to simulate subnuclear DNA chromatin fibers (CFs) of 30nm utilizing a mean-free-path approach common to radiation transport. The cell nucleus was modeled as a spherical region containing 6000 chromatin-dense domains (CDs) of 400nm diameter, with additional CFs modeled in a sparser interchromatin region. The Geant4-DNA code was utilized to produce a particle track database representing various particles at different energies and dose quantities. These tracks were used to stochastically position the DNA structures based on their mean free path to interaction with CFs. Excitation and ionization events intersecting CFs were analyzed using the DBSCAN clustering algorithm for assessment of the likelihood of producing DSBs. Simulated DSBs were then assessed based on their proximity to one another for a probability of inducing cell death. Results: Variations in energy deposition to chromatin fibers match expectations based on differences in particle track structure. The quality of damage to CFs based on different particle types indicate more severe damage by high-LET radiation than low-LET radiation of identical particles. In addition, the model indicates more severe damage by protons than of alpha particles of same LET, which is consistent with differences in their track structure. Cell survival curves have been produced showing the L-Q behavior of sparsely ionizing radiation. Conclusion: Initial results indicate the feasibility of producing cell survival curves based on the Monte Carlo cell nucleus method. Accurate correlation between simulated DNA damage to cell survival on the basis of nanodosimetric analysis can provide insight into the biological responses to various radiation types. Current efforts are directed at producing cell

  1. A systematic investigation of differential effects of cell culture substrates on the extent of artifacts in single-molecule tracking.

    Laura C Zanetti-Domingues

    Full Text Available Single-molecule techniques are being increasingly applied to biomedical investigation, notwithstanding the numerous challenges they pose in terms of signal-to-noise ratio issues. Non-specific binding of probes to glass substrates, in particular, can produce experimental artifacts due to spurious molecules on glass, which can be particularly deleterious in live-cell tracking experiments. In order to resolve the issue of non-specific probe binding to substrates, we performed systematic testing of a range of available surface coatings, using three different proteins, and then extended our assessment to the ability of these coatings to foster cell growth and retain non-adhesive properties. Linear PEG, a passivating agent commonly used both in immobilized-molecule single-molecule techniques and in tissue engineering, is able to both successfully repel non-specific adhesion of fluorescent probes and to foster cell growth when functionalized with appropriate adhesive peptides. Linear PEG treatment results in a significant reduction of tracking artifacts in EGFR tracking with Affibody ligands on a cell line expressing EGFR-eGFP. The findings reported herein could be beneficial to a large number of experimental situations where single-molecule or single-particle precision is required.

  2. Use of trimetasphere metallofullerene MRI contrast agent for the non-invasive longitudinal tracking of stem cells in the lung.

    Murphy, Sean V; Hale, Austin; Reid, Tanya; Olson, John; Kidiyoor, Amritha; Tan, Josh; Zhou, Zhiguo; Jackson, John; Atala, Anthony

    2016-04-15

    Magnetic Resonance Imaging (MRI) is a commonly used, non-invasive imaging technique that provides visualization of soft tissues with high spatial resolution. In both a research and clinical setting, the major challenge has been identifying a non-invasive and safe method for longitudinal tracking of delivered cells in vivo. The labeling and tracking of contrast agent labeled cells using MRI has the potential to fulfill this need. Contrast agents are often used to enhance the image contrast between the tissue of interest and surrounding tissues with MRI. The most commonly used MRI contrast agents contain Gd(III) ions. However, Gd(III) ions are highly toxic in their ionic form, as they tend to accumulate in the liver, spleen, kidney and bones and block calcium channels. Endohedral metallofullerenes such as trimetallic nitride endohedral metallofullerenes (Trimetasphere®) are one unique class of fullerene molecules where a Gd3N cluster is encapsulated inside a C80 carbon cage referred to as Gd3N@C80. These endohedral metallofullerenes have several advantages over small chelated Gd(III) complexes such as increased stability of the Gd(III) ion, minimal toxic effects, high solubility in water and high proton relativity. In this study, we describe the evaluation of gadolinium-based Trimetasphere® positive contrast agent for the ​in vitro labeling and in vivo tracking of human amniotic fluid-derived stem cells within lung tissue. In addition, we conducted a 'proof-of-concept' experiment demonstrating that this methodology can be used to track the homing of stem cells to injured lung tissue and provide longitudinal analysis of cell localization over an extended time course. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. In vivo cell tracking imaging of hexadecyl-4-[{sup 123,} {sup 124}I]iodobenzoate labeled adipose derived stem cells (ADSCs) in rat heart

    Kim, Min Hwan; Lee, Yong Jin; Lee, Kyo Chul [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2011-10-15

    Monitoring of transplanted stem cells for cardiac repair is important part in regenerative medicine. Direct cell labeling techniques using [{sup 18}F]FDG, [{sup 64}Cu]PTSM and [{sup 99m}Tc]-HMPAO have been developed for in vivo imaging. Especially, {sup 18}F-labeled derivates have been widely used for direct labeling agent. But the {sup 18}F has short half life (T{sub 1/2}={approx}2 h), thus this imaging agent has limitation of in vivo imaging. We used {sup 123}I or {sup 124}I which has relative long half life, to track the transplanted stem cells for a long-term imaging. This study is aimed to track the transplanted adipose derived stem cells (ADSCs) in rat heart using hexadecyl-4-[{sup 123,} {sup 124}I]iodobenzoate ([{sup 123,} {sup 124}I]HIB) mediated direct labeling method in vivo

  4. [{sup 89}Zr]Oxinate{sub 4} for long-term in vivo cell tracking by positron emission tomography

    Charoenphun, Putthiporn; Meszaros, Levente K.; Chuamsaamarkkee, Krisanat; Sharif-Paghaleh, Ehsan; Ballinger, James R.; Mullen, Gregory E.D. [St Thomas' Hospital, King' s College London, Division of Imaging Sciences and Biomedical Engineering, London (United Kingdom); Ferris, Trevor J.; Went, Michael J. [University of Kent, School of Physical Sciences, Canterbury (United Kingdom); Blower, Philip J. [St Thomas' Hospital, King' s College London, Division of Imaging Sciences and Biomedical Engineering, London (United Kingdom); King' s College London, Division of Chemistry, London (United Kingdom)

    2014-10-31

    {sup 111}In (typically as [{sup 111}In]oxinate{sub 3}) is a gold standard radiolabel for cell tracking in humans by scintigraphy. A long half-life positron-emitting radiolabel to serve the same purpose using positron emission tomography (PET) has long been sought. We aimed to develop an {sup 89}Zr PET tracer for cell labelling and compare it with [{sup 111}In]oxinate{sub 3} single photon emission computed tomography (SPECT). [{sup 89}Zr]Oxinate{sub 4} was synthesised and its uptake and efflux were measured in vitro in three cell lines and in human leukocytes. The in vivo biodistribution of eGFP-5T33 murine myeloma cells labelled using [{sup 89}Zr]oxinate{sub 4} or [{sup 111}In]oxinate{sub 3} was monitored for up to 14 days. {sup 89}Zr retention by living radiolabelled eGFP-positive cells in vivo was monitored by FACS sorting of liver, spleen and bone marrow cells followed by gamma counting. Zr labelling was effective in all cell types with yields comparable with {sup 111}In labelling. Retention of {sup 89}Zr in cells in vitro after 24 h was significantly better (range 71 to >90 %) than {sup 111}In (43-52 %). eGFP-5T33 cells in vivo showed the same early biodistribution whether labelled with {sup 111}In or {sup 89}Zr (initial pulmonary accumulation followed by migration to liver, spleen and bone marrow), but later translocation of radioactivity to kidneys was much greater for {sup 111}In. In liver, spleen and bone marrow at least 92 % of {sup 89}Zr remained associated with eGFP-positive cells after 7 days in vivo. [{sup 89}Zr]Oxinate{sub 4} offers a potential solution to the emerging need for a long half-life PET tracer for cell tracking in vivo and deserves further evaluation of its effects on survival and behaviour of different cell types. (orig.)

  5. Tracking and Quantifying Developmental Processes in C. elegans Using Open-source Tools.

    Dutta, Priyanka; Lehmann, Christina; Odedra, Devang; Singh, Deepika; Pohl, Christian

    2015-12-16

    Quantitatively capturing developmental processes is crucial to derive mechanistic models and key to identify and describe mutant phenotypes. Here protocols are presented for preparing embryos and adult C. elegans animals for short- and long-term time-lapse microscopy and methods for tracking and quantification of developmental processes. The methods presented are all based on C. elegans strains available from the Caenorhabditis Genetics Center and on open-source software that can be easily implemented in any laboratory independently of the microscopy system used. A reconstruction of a 3D cell-shape model using the modelling software IMOD, manual tracking of fluorescently-labeled subcellular structures using the multi-purpose image analysis program Endrov, and an analysis of cortical contractile flow using PIVlab (Time-Resolved Digital Particle Image Velocimetry Tool for MATLAB) are shown. It is discussed how these methods can also be deployed to quantitatively capture other developmental processes in different models, e.g., cell tracking and lineage tracing, tracking of vesicle flow.

  6. High performance monolithic power management system with dynamic maximum power point tracking for microbial fuel cells.

    Erbay, Celal; Carreon-Bautista, Salvador; Sanchez-Sinencio, Edgar; Han, Arum

    2014-12-02

    Microbial fuel cell (MFC) that can directly generate electricity from organic waste or biomass is a promising renewable and clean technology. However, low power and low voltage output of MFCs typically do not allow directly operating most electrical applications, whether it is supplementing electricity to wastewater treatment plants or for powering autonomous wireless sensor networks. Power management systems (PMSs) can overcome this limitation by boosting the MFC output voltage and managing the power for maximum efficiency. We present a monolithic low-power-consuming PMS integrated circuit (IC) chip capable of dynamic maximum power point tracking (MPPT) to maximize the extracted power from MFCs, regardless of the power and voltage fluctuations from MFCs over time. The proposed PMS continuously detects the maximum power point (MPP) of the MFC and matches the load impedance of the PMS for maximum efficiency. The system also operates autonomously by directly drawing power from the MFC itself without any external power. The overall system efficiency, defined as the ratio between input energy from the MFC and output energy stored into the supercapacitor of the PMS, was 30%. As a demonstration, the PMS connected to a 240 mL two-chamber MFC (generating 0.4 V and 512 μW at MPP) successfully powered a wireless temperature sensor that requires a voltage of 2.5 V and consumes power of 85 mW each time it transmit the sensor data, and successfully transmitted a sensor reading every 7.5 min. The PMS also efficiently managed the power output of a lower-power producing MFC, demonstrating that the PMS works efficiently at various MFC power output level.

  7. Design, Construction, Alignment, and Calibration of a Compact Velocimetry Experiment

    Morris I Kaufman; Robert M Malone; Brent C Frogget; David L Esquibel; Vincent T Romero; Gregory A Lare; Bart Briggs; Adam J Iverson; Daniel K Frayer; Douglas DeVore Brian Cata

    2007-01-01

    A velocimetry experiment has been designed to measure shock properties for small cylindrical metal targets (8-mm-diameter by 2-mm thick). A target is accelerated by high explosives, caught, and retrieved for later inspection. The target is expected to move at a velocity of 0.1 to 3 km/sec. The complete experiment canister is approximately 105 mm in diameter and 380 mm long. Optical velocimetry diagnostics include the Velocity Interferometer System for Any Reflector (VISAR) and Photon Doppler Velocimetry (PDV). The packaging of the velocity diagnostics is not allowed to interfere with the catchment or an X-ray imaging diagnostic. A single optical relay, using commercial lenses, collects Doppler-shifted light for both VISAR and PDV. The use of fiber optics allows measurement of point velocities on the target surface during accelerations occurring over 15 mm of travel. The VISAR operates at 532 nm and has separate illumination fibers requiring alignment. The PDV diagnostic operates at 1550 nm, but is aligned and focused at 670 nm. The VISAR and PDV diagnostics are complementary measurements and they image spots in close proximity on the target surface. Because the optical relay uses commercial glass, the axial positions of the optical fibers for PDV and VISAR are offset to compensate for chromatic aberrations. The optomechanical design requires careful attention to fiber management, mechanical assembly and disassembly, positioning of the foam catchment, and X-ray diagnostic field-of-view. Calibration and alignment data are archived at each stage of the assembly sequence

  8. Hybrid catadioptric system for advanced optical cavity velocimetry

    Frayer, Daniel K.

    2018-02-06

    A probe including reflector is disclosed to measure the velocity distribution of a moving surface along many lines of sight. Laser light, directed to the surface by the probe and then reflected back from the surface, is Doppler shifted by the moving surface, collected into probe, and then directed to detection equipment through optic fibers. The received light is mixed with reference laser light and using photonic Doppler velocimetry, a continuous time record of the surface movement is obtained. An array of single-mode optical fibers provides an optic signal to one or more lens groups and a reflector, such as a parabolic reflector having a mirrored interior surface.

  9. Particle and speckle imaging velocimetry applied to a monostatic LIDAR

    Halldorsson, Thorsteinn; Langmeier, Andreas; Prücklmeier, Andreas; Banakh, Viktor; Falits, Andrey

    2006-11-01

    A novel backscatter-lidar imaging method of visualization of air movement in the atmosphere is discussed in the paper. The method is based on the particle image velocimetry (PIV) principle, namely: pairs of image of laser illuminated thin atmospheric layers are recorded by CCD camera and then are cross correlated to obtain velocity information from these records. Both the way of computer simulation of atmospheric version of PIV technique and the first concept proof experiments are described in the paper. It is proposed that the method can find an application for visualization of wake vortices arising behind large aircrafts.

  10. Multiparticle imaging technique for two-phase fluid flows using pulsed laser speckle velocimetry

    Hassan, T.A.

    1992-12-01

    The practical use of Pulsed Laser Velocimetry (PLV) requires the use of fast, reliable computer-based methods for tracking numerous particles suspended in a fluid flow. Two methods for performing tracking are presented. One method tracks a particle through multiple sequential images (minimum of four required) by prediction and verification of particle displacement and direction. The other method, requiring only two sequential images uses a dynamic, binary, spatial, cross-correlation technique. The algorithms are tested on computer-generated synthetic data and experimental data which was obtained with traditional PLV methods. This allowed error analysis and testing of the algorithms on real engineering flows. A novel method is proposed which eliminates tedious, undersirable, manual, operator assistance in removing erroneous vectors. This method uses an iterative process involving an interpolated field produced from the most reliable vectors. Methods are developed to allow fast analysis and presentation of sets of PLV image data. Experimental investigation of a two-phase, horizontal, stratified, flow regime was performed to determine the interface drag force, and correspondingly, the drag coefficient. A horizontal, stratified flow test facility using water and air was constructed to allow interface shear measurements with PLV techniques. The experimentally obtained local drag measurements were compared with theoretical results given by conventional interfacial drag theory. Close agreement was shown when local conditions near the interface were similar to space-averaged conditions. However, theory based on macroscopic, space-averaged flow behavior was shown to give incorrect results if the local gas velocity near the interface as unstable, transient, and dissimilar from the average gas velocity through the test facility.

  11. In vitro and in vivo imaging and tracking of intestinal organoids from human induced pluripotent stem cells.

    Jung, Kwang Bo; Lee, Hana; Son, Ye Seul; Lee, Ji Hye; Cho, Hyun-Soo; Lee, Mi-Ok; Oh, Jung-Hwa; Lee, Jaemin; Kim, Seokho; Jung, Cho-Rok; Kim, Janghwan; Son, Mi-Young

    2018-01-01

    Human intestinal organoids (hIOs) derived from human pluripotent stem cells (hPSCs) have immense potential as a source of intestines. Therefore, an efficient system is needed for visualizing the stage of intestinal differentiation and further identifying hIOs derived from hPSCs. Here, 2 fluorescent biosensors were developed based on human induced pluripotent stem cell (hiPSC) lines that stably expressed fluorescent reporters driven by intestine-specific gene promoters Krüppel-like factor 5 monomeric Cherry (KLF5 mCherry ) and intestine-specific homeobox enhanced green fluorescence protein (ISX eGFP ). Then hIOs were efficiently induced from those transgenic hiPSC lines in which mCherry- or eGFP-expressing cells, which appeared during differentiation, could be identified in intact living cells in real time. Reporter gene expression had no adverse effects on differentiation into hIOs and proliferation. Using our reporter system to screen for hIO differentiation factors, we identified DMH1 as an efficient substitute for Noggin. Transplanted hIOs under the kidney capsule were tracked with fluorescence imaging (FLI) and confirmed histologically. After orthotopic transplantation, the localization of the hIOs in the small intestine could be accurately visualized using FLI. Our study establishes a selective system for monitoring the in vitro differentiation and for tracking the in vivo localization of hIOs and contributes to further improvement of cell-based therapies and preclinical screenings in the intestinal field.-Jung, K. B., Lee, H., Son, Y. S., Lee, J. H., Cho, H.-S., Lee, M.-O., Oh, J.-H., Lee, J., Kim, S., Jung, C.-R., Kim, J., Son, M.-Y. In vitro and in vivo imaging and tracking of intestinal organoids from human induced pluripotent stem cells. © FASEB.

  12. Multipurpose Pressure Vessel Scanner and Photon Doppler Velocimetry

    Ellis, Tayera

    2015-01-01

    Critical flight hardware typically undergoes a series of nondestructive evaluation methods to screen for defects before it is integrated into the flight system. Conventionally, pressure vessels have been inspected for flaws using a technique known as fluorescent dye penetrant, which is biased to inspector interpretation. An alternate method known as eddy current is automated and can detect small cracks better than dye penetrant. A new multipurpose pressure vessel scanner has been developed to perform internal and external eddy current scanning, laser profilometry, and thickness mapping on pressure vessels. Before this system can be implemented throughout industry, a probability of detection (POD) study needs to be performed to validate the system’s eddy current crack/flaw capabilities. The POD sample set will consist of 6 flight-like metal pressure vessel liners with defects of known size. Preparation for the POD includes sample set fabrication, system operation, procedure development, and eddy current settings optimization. For this, collaborating with subject matter experts was required. This technical paper details the preparation activities leading up to the POD study currently scheduled for winter 2015/2016. Once validated, this system will be a proven innovation for increasing the safety and reliability of necessary flight hardware.Additionally, testing of frangible joint requires Photon Doppler Velocimetry (PDV) and Digital Image Correlation instrumentation. There is often noise associated with PDV data, which necessitates a frequency modulation (FM) signal-to-noise pre-test. Generally, FM radio works by varying the carrier frequency and mixing it with a fixed frequency source, creating a beat frequency which is represented by audio frequency that can be heard between about 20 to 20,000 Hz. Similarly, PDV reflects a shifted frequency (a phenomenon known as the Doppler Effect) from a moving source and mixes it with a fixed source frequency, which results in

  13. Explicit tracking of uncertainty increases the power of quantitative rule-of-thumb reasoning in cell biology.

    Johnston, Iain G; Rickett, Benjamin C; Jones, Nick S

    2014-12-02

    Back-of-the-envelope or rule-of-thumb calculations involving rough estimates of quantities play a central scientific role in developing intuition about the structure and behavior of physical systems, for example in so-called Fermi problems in the physical sciences. Such calculations can be used to powerfully and quantitatively reason about biological systems, particularly at the interface between physics and biology. However, substantial uncertainties are often associated with values in cell biology, and performing calculations without taking this uncertainty into account may limit the extent to which results can be interpreted for a given problem. We present a means to facilitate such calculations where uncertainties are explicitly tracked through the line of reasoning, and introduce a probabilistic calculator called CALADIS, a free web tool, designed to perform this tracking. This approach allows users to perform more statistically robust calculations in cell biology despite having uncertain values, and to identify which quantities need to be measured more precisely to make confident statements, facilitating efficient experimental design. We illustrate the use of our tool for tracking uncertainty in several example biological calculations, showing that the results yield powerful and interpretable statistics on the quantities of interest. We also demonstrate that the outcomes of calculations may differ from point estimates when uncertainty is accurately tracked. An integral link between CALADIS and the BioNumbers repository of biological quantities further facilitates the straightforward location, selection, and use of a wealth of experimental data in cell biological calculations. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  14. Hybrid micro-/nano-particle image velocimetry for 3D3C multi-scale velocity field measurement in microfluidics

    Min, Young Uk; Kim, Kyung Chun

    2011-01-01

    The conventional two-dimensional (2D) micro-particle image velocimetry (micro-PIV) technique has inherent bias error due to the depth of focus along the optical axis to measure the velocity field near the wall of a microfluidics device. However, the far-field measurement of velocity vectors yields good accuracy for micro-scale flows. Nano-PIV using the evanescent wave of total internal reflection fluorescence microscopy can measure near-field velocity vectors within a distance of around 200 nm from the solid surface. A micro-/nano-hybrid PIV system is proposed to measure both near- and far-field velocity vectors simultaneously in microfluidics. A near-field particle image can be obtained by total internal reflection fluorescence microscopy using nanoparticles, and the far-field velocity vectors are measured by three-hole defocusing micro-particle tracking velocimetry (micro-PTV) using micro-particles. In order to identify near- and far-field particle images, lasers of different wavelengths are adopted and tested in a straight microchannel for acquiring the three-dimensional three-component velocity field. We found that the new technique gives superior accuracy for the velocity profile near the wall compared to that of conventional nano-PIV. This method has been successfully applied to precisely measure wall shear stress in 2D microscale Poiseulle flows

  15. Tomographic Particle Image Velocimetry Using Colored Shadow Imaging

    Alarfaj, Meshal K.

    2016-02-01

    Tomographic Particle Image Velocimetry Using Colored Shadow Imaging by Meshal K Alarfaj, Master of Science King Abdullah University of Science & Technology, 2015 Tomographic Particle image velocimetry (PIV) is a recent PIV method capable of reconstructing the full 3D velocity field of complex flows, within a 3-D volume. For nearly the last decade, it has become the most powerful tool for study of turbulent velocity fields and promises great advancements in the study of fluid mechanics. Among the early published studies, a good number of researches have suggested enhancements and optimizations of different aspects of this technique to improve the effectiveness. One major aspect, which is the core of the present work, is related to reducing the cost of the Tomographic PIV setup. In this thesis, we attempt to reduce this cost by using an experimental setup exploiting 4 commercial digital still cameras in combination with low-cost Light emitting diodes (LEDs). We use two different colors to distinguish the two light pulses. By using colored shadows with red and green LEDs, we can identify the particle locations within the measurement volume, at the two different times, thereby allowing calculation of the velocities. The present work tests this technique on the flows patterns of a jet ejected from a tube in a water tank. Results from the images processing are presented and challenges discussed.

  16. Systematic Error of Acoustic Particle Image Velocimetry and Its Correction

    Mickiewicz Witold

    2014-08-01

    Full Text Available Particle Image Velocimetry is getting more and more often the method of choice not only for visualization of turbulent mass flows in fluid mechanics, but also in linear and non-linear acoustics for non-intrusive visualization of acoustic particle velocity. Particle Image Velocimetry with low sampling rate (about 15Hz can be applied to visualize the acoustic field using the acquisition synchronized to the excitation signal. Such phase-locked PIV technique is described and used in experiments presented in the paper. The main goal of research was to propose a model of PIV systematic error due to non-zero time interval between acquisitions of two images of the examined sound field seeded with tracer particles, what affects the measurement of complex acoustic signals. Usefulness of the presented model is confirmed experimentally. The correction procedure, based on the proposed model, applied to measurement data increases the accuracy of acoustic particle velocity field visualization and creates new possibilities in observation of sound fields excited with multi-tonal or band-limited noise signals.

  17. Noise Studies of Externally Dispersed Interferometry for Doppler Velocimetry

    Erskine, D J; Edelstein, J; Lloyd, J; Muirhead, P

    2006-01-01

    Externally Dispersed Interferometry (EDI) is the series combination of a fixed-delay field-widened Michelson interferometer with a dispersive spectrograph. This combination boosts the spectrograph performance for both Doppler velocimetry and high resolution spectroscopy. The interferometer creates a periodic comb that multiplies against the input spectrum to create moire fringes, which are recorded in combination with the regular spectrum. Both regular and high-frequency spectral components can be recovered from the data--the moire component carries additional information that increases the signal to noise for velocimetry and spectroscopy. Here we present simulations and theoretical studies of the photon limited Doppler velocity noise in an EDI. We used a model spectrum of a 1600K temperature star. For several rotational blurring velocities 0, 7.5, 15 and 25 km/s we calculated the dimensionless Doppler quality index (Q) versus wavenumber v. This is the normalized RMS of the derivative of the spectrum and is proportional to the photon-limited Doppler signal to noise ratio

  18. The NASA Subsonic Jet Particle Image Velocimetry (PIV) Dataset

    Bridges, James; Wernet, Mark P.

    2011-01-01

    Many tasks in fluids engineering require prediction of turbulence of jet flows. The present document documents the single-point statistics of velocity, mean and variance, of cold and hot jet flows. The jet velocities ranged from 0.5 to 1.4 times the ambient speed of sound, and temperatures ranged from unheated to static temperature ratio 2.7. Further, the report assesses the accuracies of the data, e.g., establish uncertainties for the data. This paper covers the following five tasks: (1) Document acquisition and processing procedures used to create the particle image velocimetry (PIV) datasets. (2) Compare PIV data with hotwire and laser Doppler velocimetry (LDV) data published in the open literature. (3) Compare different datasets acquired at the same flow conditions in multiple tests to establish uncertainties. (4) Create a consensus dataset for a range of hot jet flows, including uncertainty bands. (5) Analyze this consensus dataset for self-consistency and compare jet characteristics to those of the open literature. The final objective was fulfilled by using the potential core length and the spread rate of the half-velocity radius to collapse of the mean and turbulent velocity fields over the first 20 jet diameters.

  19. Analysis of particle kinematics in spheronization via particle image velocimetry.

    Koester, Martin; Thommes, Markus

    2013-02-01

    Spheronization is a wide spread technique in pellet production for many pharmaceutical applications. Pellets produced by spheronization are characterized by a particularly spherical shape and narrow size distribution. The particle kinematic during spheronization is currently not well-understood. Therefore, particle image velocimetry (PIV) was implemented in the spheronization process to visualize the particle movement and to identify flow patterns, in order to explain the influence of various process parameters. The spheronization process of a common formulation was recorded with a high-speed camera, and the images were processed using particle image velocimetry software. A crosscorrelation approach was chosen to determine the particle velocity at the surface of the pellet bulk. Formulation and process parameters were varied systematically, and their influence on the particle velocity was investigated. The particle stream shows a torus-like shape with a twisted rope-like motion. It is remarkable that the overall particle velocity is approximately 10-fold lower than the tip speed of the friction plate. The velocity of the particle stream can be correlated to the water content of the pellets and the load of the spheronizer, while the rotation speed was not relevant. In conclusion, PIV was successfully applied to the spheronization process, and new insights into the particle velocity were obtained. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Impact of time and space evolution of ion tracks in nonvolatile memory cells approaching nanoscale

    Cellere, G.; Paccagnella, A.; Murat, M.; Barak, J.; Akkerman, A.; Harboe-Sorensen, R.; Virtanen, A.; Visconti, A.; Bonanomi, M.

    2010-01-01

    Swift heavy ions impacting on matter lose energy through the creation of dense tracks of charges. The study of the space and time evolution of energy exchange allows understanding the single event effects behavior in advanced microelectronic devices. In particular, the shrinking of minimum feature size of most advanced memory devices makes them very interesting test vehicles to study these effects since the device and the track dimensions are comparable; hence, measured effects are directly correlated with the time and space evolution of the energy release. In this work we are studying the time and space evolution of ion tracks by using advanced non volatile memories and Monte Carlo simulations. Experimental results are very well explained by the theoretical calculations.

  1. Tracking of cell nuclei for assessment of in vitro uptake kinetics in ultrasound-mediated drug delivery using fibered confocal fluorescence microscopy.

    Derieppe, Marc; de Senneville, Baudouin Denis; Kuijf, Hugo; Moonen, Chrit; Bos, Clemens

    2014-10-01

    Previously, we demonstrated the feasibility to monitor ultrasound-mediated uptake of a cell-impermeable model drug in real time with fibered confocal fluorescence microscopy. Here, we present a complete post-processing methodology, which corrects for cell displacements, to improve the accuracy of pharmacokinetic parameter estimation. Nucleus detection was performed based on the radial symmetry transform algorithm. Cell tracking used an iterative closest point approach. Pharmacokinetic parameters were calculated by fitting a two-compartment model to the time-intensity curves of individual cells. Cells were tracked successfully, improving time-intensity curve accuracy and pharmacokinetic parameter estimation. With tracking, 93 % of the 370 nuclei showed a fluorescence signal variation that was well-described by a two-compartment model. In addition, parameter distributions were narrower, thus increasing precision. Dedicated image analysis was implemented and enabled studying ultrasound-mediated model drug uptake kinetics in hundreds of cells per experiment, using fiber-based confocal fluorescence microscopy.

  2. Krypton tagging velocimetry in a turbulent Mach 2.7 boundary layer

    Zahradka, D.; Parziale, N. J.; Smith, M. S.; Marineau, E. C.

    2016-05-01

    The krypton tagging velocimetry (KTV) technique is applied to the turbulent boundary layer on the wall of the "Mach 3 Calibration Tunnel" at Arnold Engineering Development Complex (AEDC) White Oak. Profiles of velocity were measured with KTV and Pitot-pressure probes in the Mach 2.7 turbulent boundary layer comprised of 99 % {N}2/1 % Kr at momentum-thickness Reynolds numbers of {Re}_{\\varTheta }= 800, 1400, and 2400. Agreement between the KTV- and Pitot-derived velocity profiles is excellent. The KTV and Pitot velocity data follow the law of the wall in the logarithmic region with application of the Van Driest I transformation. The velocity data are analyzed in the outer region of the boundary layer with the law of the wake and a velocity-defect law. KTV-derived streamwise velocity fluctuation measurements are reported and are consistent with data from the literature. To enable near-wall measurement with KTV (y/δ ≈ 0.1-0.2), an 800-nm longpass filter was used to block the 760.2-nm read-laser pulse. With the longpass filter, the 819.0-nm emission from the re-excited Kr can be imaged to track the displacement of the metastable tracer without imaging the reflection and scatter from the read-laser off of solid surfaces. To operate the Mach 3 AEDC Calibration Tunnel at several discrete unit Reynolds numbers, a modification was required and is described herein.

  3. Investigation and visualization of internal flow through particle aggregates and microbial flocs using particle image velocimetry.

    Xiao, Feng; Lam, Kit Ming; Li, Xiao-yan

    2013-05-01

    An advanced particle-tracking and flow-visualization technology, particle image velocimetry (PIV), was utilized to investigate the hydrodynamic properties of large aggregates in water. The laser-based PIV system was used together with a settling column to capture the streamlines around two types of aggregates: latex particle aggregates and activated sludge (AS) flocs. Both types of the aggregates were highly porous and fractal with fractal dimensions of 2.13±0.31 for the latex particle aggregates (1210-2144 μm) and 1.78±0.24 for the AS flocs (1265-3737 μm). The results show that PIV is a powerful flow visualization technique capable of determining flow field details at the micrometer scale around and through settling aggregates and flocs. The PIV streamlines provided direct experimental proof of internal flow through the aggregate interiors. According to the PIV images, fluid collection efficiency ranged from 0.052 to 0.174 for the latex particle aggregates and from 0.008 to 0.126 for AS flocs. AS flocs are apparently less permeable than the particle aggregates, probably due to the extracellular polymeric substances (EPSs) produced by bacteria clogging the pores within the flocs. The internal permeation of fractal aggregates and bio-flocs would enhance flocculation between particles and material transport into the aggregates. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. An automated approach for single-cell tracking in epifluorescence microscopy applied to E. coli growth analysis on microfluidics biochips

    Fetita, Catalin; Kirov, Boris; Jaramillo, Alfonso; Lefevre, Christophe

    2012-03-01

    With the accumulation of knowledge for the intimate molecular mechanisms governing the processes inside the living cells in the later years, the ability to characterize the performance of elementary genetic circuits and parts at the single-cell level is becoming of crucial importance. Biological science is arriving to the point where it can develop hypothesis for the action of each molecule participating in the biochemical reactions and need proper techniques to test those hypothesis. Microfluidics is emerging as the technology that combined with high-magnification microscopy will allow for the long-term single-cell level observation of bacterial physiology. In this study we design, build and characterize the gene dynamics of genetic circuits as one of the basic parts governing programmed cell behavior. We use E. coli as model organism and grow it in microfluidics chips, which we observe with epifluorescence microscopy. One of the most invaluable segments of this technology is the consequent image processing, since it allows for the automated analysis of vast amount of single-cell observation and the fast and easy derivation of conclusions based on that data. Specifically, we are interested in promoter activity as function of time. We expect it to be oscillatory and for that we use GFP (green fluorescent protein) as a reporter in our genetic circuits. In this paper, an automated framework for single-cell tracking in phase-contrast microscopy is developed, combining 2D segmentation of cell time frames and graph-based reconstruction of their spatiotemporal evolution with fast tracking of the associated fluorescence signal. The results obtained on the investigated biological database are presented and discussed.

  5. Hail statistic in Western Europe based on a hyrid cell-tracking algorithm combining radar signals with hailstone observations

    Fluck, Elody

    2015-04-01

    Hail statistic in Western Europe based on a hybrid cell-tracking algorithm combining radar signals with hailstone observations Elody Fluck¹, Michael Kunz¹ , Peter Geissbühler², Stefan P. Ritz² With hail damage estimated over Billions of Euros for a single event (e.g., hailstorm Andreas on 27/28 July 2013), hail constitute one of the major atmospheric risks in various parts of Europe. The project HAMLET (Hail Model for Europe) in cooperation with the insurance company Tokio Millennium Re aims at estimating hail probability, hail hazard and, combined with vulnerability, hail risk for several European countries (Germany, Switzerland, France, Netherlands, Austria, Belgium and Luxembourg). Hail signals are obtained from radar reflectivity since this proxy is available with a high temporal and spatial resolution using several hail proxies, especially radar data. The focus in the first step is on Germany and France for the periods 2005- 2013 and 1999 - 2013, respectively. In the next step, the methods will be transferred and extended to other regions. A cell-tracking algorithm TRACE2D was adjusted and applied to two dimensional radar reflectivity data from different radars operated by European weather services such as German weather service (DWD) and French weather service (Météo-France). Strong convective cells are detected by considering 3 connected pixels over 45 dBZ (Reflectivity Cores RCs) in a radar scan. Afterwards, the algorithm tries to find the same RCs in the next 5 minute radar scan and, thus, track the RCs centers over time and space. Additional information about hailstone diameters provided by ESWD (European Severe Weather Database) is used to determine hail intensity of the detected hail swaths. Maximum hailstone diameters are interpolated along and close to the individual hail tracks giving an estimation of mean diameters for the detected hail swaths. Furthermore, a stochastic event set is created by randomizing the parameters obtained from the

  6. Enhanced object-based tracking algorithm for convective rain storms and cells

    Muñoz, Carlos; Wang, Li-Pen; Willems, Patrick

    2018-03-01

    This paper proposes a new object-based storm tracking algorithm, based upon TITAN (Thunderstorm Identification, Tracking, Analysis and Nowcasting). TITAN is a widely-used convective storm tracking algorithm but has limitations in handling small-scale yet high-intensity storm entities due to its single-threshold identification approach. It also has difficulties to effectively track fast-moving storms because of the employed matching approach that largely relies on the overlapping areas between successive storm entities. To address these deficiencies, a number of modifications are proposed and tested in this paper. These include a two-stage multi-threshold storm identification, a new formulation for characterizing storm's physical features, and an enhanced matching technique in synergy with an optical-flow storm field tracker, as well as, according to these modifications, a more complex merging and splitting scheme. High-resolution (5-min and 529-m) radar reflectivity data for 18 storm events over Belgium are used to calibrate and evaluate the algorithm. The performance of the proposed algorithm is compared with that of the original TITAN. The results suggest that the proposed algorithm can better isolate and match convective rainfall entities, as well as to provide more reliable and detailed motion estimates. Furthermore, the improvement is found to be more significant for higher rainfall intensities. The new algorithm has the potential to serve as a basis for further applications, such as storm nowcasting and long-term stochastic spatial and temporal rainfall generation.

  7. TLM-Tracker: software for cell segmentation, tracking and lineage analysis in time-lapse microscopy movies.

    Klein, Johannes; Leupold, Stefan; Biegler, Ilona; Biedendieck, Rebekka; Münch, Richard; Jahn, Dieter

    2012-09-01

    Time-lapse imaging in combination with fluorescence microscopy techniques enable the investigation of gene regulatory circuits and uncovered phenomena like culture heterogeneity. In this context, computational image processing for the analysis of single cell behaviour plays an increasing role in systems biology and mathematical modelling approaches. Consequently, we developed a software package with graphical user interface for the analysis of single bacterial cell behaviour. A new software called TLM-Tracker allows for the flexible and user-friendly interpretation for the segmentation, tracking and lineage analysis of microbial cells in time-lapse movies. The software package, including manual, tutorial video and examples, is available as Matlab code or executable binaries at http://www.tlmtracker.tu-bs.de.

  8. Long-term MRI tracking of dual-labeled adipose-derived stem cells homing into mouse carotid artery injury

    Qin JB

    2012-10-01

    Full Text Available Jin-Bao Qin,1,5,* Kang-An Li,2,* Xiang-Xiang Li,1,5 Qing-Song Xie,3 Jia-Ying Lin,4 Kai-Chuang Ye,1,5 Mi-Er Jiang,1,5 Gui-Xiang Zhang,2 Xin-Wu Lu1,51Department of Vascular Surgery, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 2Department of Radiology, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 3Department of Neurosurgery, Cixi Municipal People's Hospital, Zhejiang Province, China; 4Clinic for Gynecology, Charite-Universitatsmedizin Berlin, Berlin, Germany; 5Vascular Center, Shanghai Jiao Tong University, Shanghai, China*These two authors contributed equally to this workBackground: Stem cell therapy has shown great promise for regenerative repair of injured or diseased tissues. Adipose-derived stem cells (ADSCs have become increasingly attractive candidates for cellular therapy. Magnetic resonance imaging has been proven to be effective in tracking magnetic-labeled cells and evaluating their clinical relevance after cell transplantation. This study investigated the feasibility of imaging green fluorescent protein-expressing ADSCs (GFP-ADSCs labeled with superparamagnetic iron oxide particles, and tracked them in vivo with noninvasive magnetic resonance imaging after cell transplantation in a model of mouse carotid artery injury.Methods: GFP-ADSCs were isolated from the adipose tissues of GFP mice and labeled with superparamagnetic iron oxide particles. Intracellular stability, proliferation, and viability of the labeled cells were evaluated in vitro. Next, the cells were transplanted into a mouse carotid artery injury model. Clinical 3 T magnetic resonance imaging was performed immediately before and 1, 3, 7, 14, 21, and 30 days after cell transplantation. Prussian blue staining and histological analysis were performed 7 and 30 days after transplantation.Results: GFP-ADSCs were found to be efficiently labeled with superparamagnetic iron oxide

  9. Simulation of DNA Damage in Human Cells from Space Radiation Using a Physical Model of Stochastic Particle Tracks and Chromosomes

    Ponomarev, Artem; Plante, Ianik; Hada, Megumi; George, Kerry; Wu, Honglu

    2015-01-01

    The formation of double-strand breaks (DSBs) and chromosomal aberrations (CAs) is of great importance in radiation research and, specifically, in space applications. We are presenting a recently developed model, in which chromosomes simulated by NASARTI (NASA Radiation Tracks Image) is combined with nanoscopic dose calculations performed with the Monte-Carlo simulation by RITRACKS (Relativistic Ion Tracks) in a voxelized space. The model produces the number of DSBs, as a function of dose for high-energy iron, oxygen, and carbon ions, and He ions. The combined model calculates yields of radiation-induced CAs and unrejoined chromosome breaks in normal and repair deficient cells. The merged computational model is calibrated using the relative frequencies and distributions of chromosomal aberrations reported in the literature. The model considers fractionated deposition of energy to approximate dose rates of the space flight environment. The merged model also predicts of the yields and sizes of translocations, dicentrics, rings, and more complex-type aberrations formed in the G0/G1 cell cycle phase during the first cell division after irradiation.

  10. Physical activity measured using global positioning system tracking in non-small cell lung cancer: an observational study.

    Granger, Catherine L; Denehy, Linda; McDonald, Christine F; Irving, Louis; Clark, Ross A

    2014-11-01

    Increasingly physical activity (PA) is being recognized as an important outcome in non-small cell lung cancer (NSCLC). We investigated PA using novel global positioning system (GPS) tracking individuals with NSCLC and a group of similar-aged healthy individuals. A prospective cross-sectional multicenter study. Fifty individuals with NSCLC from 3 Australian tertiary hospitals and 35 similar-aged healthy individuals without cancer were included. Individuals with NSCLC were assessed pretreatment. Primary measures were triaxial accelerometery (steps/day) and GPS tracking (outdoor PA behavior). Secondary measures were questionnaires assessing depression, motivation to exercise, and environmental barriers to PA. Between-group comparisons were analyzed using analysis of covariance. Individuals with NSCLC engaged in significantly less PA than similar-aged healthy individuals (mean difference 2363 steps/day, P = .007) and had higher levels of depression (P = .027) and lower motivation to exercise (P = .001). Daily outdoor walking time (P = .874) and distance travelled away from home (P = .883) were not different between groups. Individuals with NSCLC spent less time outdoors in their local neighborhood area (P system tracking appears to be a feasible methodology for adult cancer patients and holds promise for use in future studies investigating PA and or lifestyle behaviors. © The Author(s) 2014.

  11. Cellular size as a means of tracking mTOR activity and cell fate of CD4+ T cells upon antigen recognition.

    Kristen N Pollizzi

    Full Text Available mTOR is a central integrator of metabolic and immunological stimuli, dictating immune cell activation, proliferation and differentiation. In this study, we demonstrate that within a clonal population of activated T cells, there exist both mTORhi and mTORlo cells exhibiting highly divergent metabolic and immunologic functions. By taking advantage of the role of mTOR activation in controlling cellular size, we demonstrate that upon antigen recognition, mTORhi CD4+ T cells are destined to become highly glycolytic effector cells. Conversely, mTORlo T cells preferentially develop into long-lived cells that express high levels of Bcl-2, CD25, and CD62L. Furthermore, mTORlo T cells have a greater propensity to differentiate into suppressive Foxp3+ T regulatory cells, and this paradigm was also observed in human CD4+ T cells. Overall, these studies provide the opportunity to track the development of effector and memory T cells from naïve precursors, as well as facilitate the interrogation of immunologic and metabolic programs that inform these fates.

  12. SU-E-T-494: Influence of Proton Track-Cell Nucleus Incidence Angle On Relative Biological Effectiveness

    Pater, P; Backstrom, G; Enger, S; Seuntjens, J; El Naqa, I [McGill University, Montreal, Quebec (Canada); Villegas, F; Ahnesjo, A [Uppsala University, Uppsala (Sweden)

    2015-06-15

    Purpose: To explain a Monte Carlo (MC) simulation artifact whereby differences in relative biological effectiveness (RBE) in the induction of initial double strand breaks are observed as a function of the proton track incidence angles in a geometric cell nucleus model. Secondly, to offer an alternative isotropic irradiation procedure to mitigate this effect. Methods: MC tracks of 1 MeV protons were generated in an event-by-event mode. They were overlaid on a cylindrical model of a cell nucleus containing 6×109 nucleotide base pairs. The tracks incidence angle θ with respect to the cell nucleus’s axis was varied in 10 degrees intervals, each time generating one hundred fractions of ∼2 Gy. Strand breaks were scored in the modeled DNA sugar-phosphate groups and further sub-classified into single or double strand breaks (ssbs or dsbs). For each angle, an RBE for the induction of initial dsbs with reference to Co-60 was calculated. Results: Our results show significant angular dependencies of RBE, with maximum values for incidence angles parallel to the nucleus central axis. Further examination shows that the higher cross-sections for the creation of dsbs is due to the preferential alignment of tracks with geometrical sub-parts of the cell nucleus model, especially the nucleosomes containing the sugar-phosphate groups. To alleviate the impact of this simulation artifact, an average RBE was calculated with a procedure based on a weighted sampling of the angular data. Conclusion: This work demonstrates a possible numerical artifact in estimated RBE if the influence of the particle incidence angle is not correctly taken into account. A correction procedure is presented to better conform the simulations to real-life experimental conditions. We would like to acknowledge support from the Fonds de recherche du Quebec Sante (FRQS), from the CREATE Medical Physics Research Training Network grant (number 432290) of NSERC, support from NSERC under grants RGPIN 397711-11 and

  13. The Microtubule Plus-End Tracking Protein CLASP2 Is Required for Hematopoiesis and Hematopoietic Stem Cell Maintenance

    Ksenija Drabek

    2012-10-01

    Full Text Available Mammalian CLASPs are microtubule plus-end tracking proteins whose essential function as regulators of microtubule behavior has been studied mainly in cultured cells. We show here that absence of murine CLASP2 in vivo results in thrombocytopenia, progressive anemia, and pancytopenia, due to defects in megakaryopoiesis, in erythropoiesis, and in the maintenance of hematopoietic stem cell activity. Furthermore, microtubule stability and organization are affected upon attachment of Clasp2 knockout hematopoietic stem-cell-enriched populations, and these cells do not home efficiently toward their bone marrow niche. Strikingly, CLASP2-deficient hematopoietic stem cells contain severely reduced mRNA levels of c-Mpl, which encodes the thrombopoietin receptor, an essential factor for megakaryopoiesis and hematopoietic stem cell maintenance. Our data suggest that thrombopoietin signaling is impaired in Clasp2 knockout mice. We propose that the CLASP2-mediated stabilization of microtubules is required for proper attachment, homing, and maintenance of hematopoietic stem cells and that this is necessary to sustain c-Mpl transcription.

  14. Infrared fluorescent protein 1.4 genetic labeling tracks engrafted cardiac progenitor cells in mouse ischemic hearts.

    Lijuan Chen

    Full Text Available Stem cell therapy has a potential for regenerating damaged myocardium. However, a key obstacle to cell therapy's success is the loss of engrafted cells due to apoptosis or necrosis in the ischemic myocardium. While many strategies have been developed to improve engrafted cell survival, tools to evaluate cell efficacy within the body are limited. Traditional genetic labeling tools, such as GFP-like fluorescent proteins (eGFP, DsRed, mCherry, have limited penetration depths in vivo due to tissue scattering and absorption. To circumvent these limitations, a near-infrared fluorescent mutant of the DrBphP bacteriophytochrome from Deinococcus radiodurans, IFP1.4, was developed for in vivo imaging, but it has yet to be used for in vivo stem/progenitor cell tracking. In this study, we incorporated IFP1.4 into mouse cardiac progenitor cells (CPCs by a lentiviral vector. Live IFP1.4-labeled CPCs were imaged by their near-infrared fluorescence (NIRF using an Odyssey scanner following overnight incubation with biliverdin. A significant linear correlation was observed between the amount of cells and NIRF signal intensity in in vitro studies. Lentiviral mediated IFP1.4 gene labeling is stable, and does not impact the apoptosis and cardiac differentiation of CPC. To assess efficacy of our model for engrafted cells in vivo, IFP1.4-labeled CPCs were intramyocardially injected into infarcted hearts. NIRF signals were collected at 1-day, 7-days, and 14-days post-injection using the Kodak in vivo multispectral imaging system. Strong NIRF signals from engrafted cells were imaged 1 day after injection. At 1 week after injection, 70% of the NIRF signal was lost when compared to the intensity of the day 1 signal. The data collected 2 weeks following transplantation showed an 88% decrease when compared to day 1. Our studies have shown that IFP1.4 gene labeling can be used to track the viability of transplanted cells in vivo.

  15. Multiparticle imaging velocimetry measurements in two-phase flow

    Hassan, Y.A.

    1998-01-01

    The experimental flow visualization tool, Particle Image Velocimetry (PIV), is being extended to determine the velocity fields in two and three-dimensional, two-phase fluid flows. In the past few years, the technique has attracted quite a lot of interest. PIV enables fluid velocities across a region of a flow to be measured at a single instant in time in global domain. This instantaneous velocity profile of a given flow field is determined by digitally recording particle (microspheres or bubbles) images within the flow over multiple successive video frames and then conducting flow pattern identification and analysis of the data. This paper presents instantaneous velocity measurements in various two and three- dimensional, two-phase flow situations. (author)

  16. Real-time particle image velocimetry based on FPGA technology

    Iriarte Munoz, Jose Miguel

    2008-01-01

    Particle image velocimetry (PIV), based on laser sheet, is a method for image processing and calculation of distributed velocity fields.It is well established as a fluid dynamics measurement tool, being applied to liquid, gases and multiphase flows.Images of particles are processed by means of computationally demanding algorithms, what makes its real-time implementation difficult.The most probable displacements are found applying two dimensional cross-correlation function. In this work, we detail how it is possible to achieve real-time visualization of PIV method by designing an adaptive embedded architecture based on FPGA technology.We show first results of a physical field of velocity calculated by this platform system in a real-time approach. [es

  17. Particle image velocimetry new developments and recent applications

    Willert, Christian E

    2008-01-01

    Particle Image Velocimetry (PIV) is a non-intrusive optical measurement technique which allows capturing several thousand velocity vectors within large flow fields instantaneously. Today, the PIV technique has spread widely and differentiated into many distinct applications, from micro flows over combustion to supersonic flows for both industrial needs and research. Over the past decade the measurement technique and the hard- and software have been improved continuously so that PIV has become a reliable and accurate method for "real life" investigations. Nevertheless there is still an ongoing process of improvements and extensions of the PIV technique towards 3D, time resolution, higher accuracy, measurements under harsh conditions and micro- and macroscales. This book gives a synopsis of the main results achieved during the EC-funded network PivNet 2 as well as a survey of the state-of-the-art of scientific research using PIV techniques in different fields of application.

  18. Digital Particle Image Velocimetry: Partial Image Error (PIE)

    Anandarajah, K; Hargrave, G K; Halliwell, N A

    2006-01-01

    This paper quantifies the errors due to partial imaging of seeding particles which occur at the edges of interrogation regions in Digital Particle Image Velocimetry (DPIV). Hitherto, in the scientific literature the effect of these partial images has been assumed to be negligible. The results show that the error is significant even at a commonly used interrogation region size of 32 x 32 pixels. If correlation of interrogation region sizes of 16 x 16 pixels and smaller is attempted, the error which occurs can preclude meaningful results being obtained. In order to reduce the error normalisation of the correlation peak values is necessary. The paper introduces Normalisation by Signal Strength (NSS) as the preferred means of normalisation for optimum accuracy. In addition, it is shown that NSS increases the dynamic range of DPIV

  19. Multipoint photonic doppler velocimetry using optical lens elements

    Frogget, Brent Copely; Romero, Vincent Todd

    2014-04-29

    A probe including a fisheye lens is disclosed to measure the velocity distribution of a moving surface along many lines of sight. Laser light, directed to the surface and then reflected back from the surface, is Doppler shifted by the moving surface, collected into fisheye lens, and then directed to detection equipment through optic fibers. The received light is mixed with reference laser light and using photonic Doppler velocimetry, a continuous time record of the surface movement is obtained. An array of single-mode optical fibers provides an optic signal to an index-matching lens and eventually to a fisheye lens. The fiber array flat polished and coupled to the index-matching lens using index-matching gel. Numerous fibers in a fiber array project numerous rays through the fisheye lens which in turn project many measurement points at numerous different locations to establish surface coverage over a hemispherical shape with very little crosstalk.

  20. New adaptive sampling method in particle image velocimetry

    Yu, Kaikai; Xu, Jinglei; Tang, Lan; Mo, Jianwei

    2015-01-01

    This study proposes a new adaptive method to enable the number of interrogation windows and their positions in a particle image velocimetry (PIV) image interrogation algorithm to become self-adapted according to the seeding density. The proposed method can relax the constraint of uniform sampling rate and uniform window size commonly adopted in the traditional PIV algorithm. In addition, the positions of the sampling points are redistributed on the basis of the spring force generated by the sampling points. The advantages include control of the number of interrogation windows according to the local seeding density and smoother distribution of sampling points. The reliability of the adaptive sampling method is illustrated by processing synthetic and experimental images. The synthetic example attests to the advantages of the sampling method. Compared with that of the uniform interrogation technique in the experimental application, the spatial resolution is locally enhanced when using the proposed sampling method. (technical design note)

  1. Characterization of extrusion flow using particle image velocimetry

    2009-09-01

    Full Text Available The aim of this study was the characterization of polymer flows within an extrusion die using particle image velocimetry (PIV in very constraining conditions (high temperature, pressure and velocity. Measurements were realized on semi-industrial equipments in order to have test conditions close to the industrial ones. Simple flows as well as disrupted ones were studied in order to determine the capabilities and the limits of the method. The analysis of the velocity profiles pointed out significant wall slip, which was confirmed by rheological measurements based on Mooney's method. Numerical simulations were used to connect the two sets of measurements and to simulate complex velocity profiles for comparison to the experimental ones. A good agreement was found between simulations and experiments providing wall slip is taken into account in the simulation.

  2. Holographic Particle Image Velocimetry and its Application in Engine Development

    Coupland, J M; Garner, C P; Alcock, R D; Halliwell, N A

    2006-01-01

    This paper reviews Holographic Particle Image Velocimetry (HPIV) as a means to make three-component velocity measurements throughout a three-dimensional flow-field of interest. A simplified treatment of three-dimensional scalar wave propagation is outlined and subsequently used to illustrate the principles of complex correlation analysis. It is shown that this type of analysis provides the three-dimensional correlation of the propagating, monochromatic fields recorded by the hologram. A similar approach is used to analyse the Object Conjugate Reconstruction (OCR) technique to resolve directional ambiguity by introducing an artificial image shift to the reconstructed particle images. An example of how these methods are used together to measure the instantaneous flow fields within a motored Diesel engine is then described

  3. Cell tracking with gadophrin-2: a bifunctional contrast agent for MR imaging, optical imaging, and fluorescence microscopy

    Daldrup-Link, Heike E.; Rudelius, Martina; Piontek, Guido; Schlegel, Juergen; Metz, Stephan; Settles, Marcus; Rummeny, Ernst J.; Pichler, Bernd; Heinzmann, Ulrich; Oostendorp, Robert A.J.

    2004-01-01

    The purpose of this study was to assess the feasibility of use of gadophrin-2 to trace intravenously injected human hematopoietic cells in athymic mice, employing magnetic resonance (MR) imaging, optical imaging (OI), and fluorescence microscopy. Mononuclear peripheral blood cells from GCSF-primed patients were labeled with gadophrin-2 (Schering AG, Berlin, Germany), a paramagnetic and fluorescent metalloporphyrin, using established transfection techniques with cationic liposomes. The labeled cells were evaluated in vitro with electron microscopy and inductively coupled plasma atomic emission spectrometry. Then, 1 x 10 6 -3 x 10 8 labeled cells were injected into 14 nude Balb/c mice and the in vivo cell distribution was evaluated with MR imaging and OI before and 4, 24, and 48 h after intravenous injection (p.i.). Five additional mice served as controls: three mice were untreated controls and two mice were investigated after injection of unlabeled cells. The contrast agent effect was determined quantitatively for MR imaging by calculating signal-to-noise-ratio (SNR) data. After completion of in vivo imaging studies, fluorescence microscopy of excised organs was performed. Intracellular cytoplasmatic uptake of gadophrin-2 was confirmed by electron microscopy. Spectrometry determined an uptake of 31.56 nmol Gd per 10 6 cells. After intravenous injection, the distribution of gadophrin-2 labeled cells in nude mice could be visualized by MR, OI, and fluorescence microscopy. At 4 h p.i., the transplanted cells mainly distributed to lung, liver, and spleen, and 24 h p.i. they also distributed to the bone marrow. Fluorescence microscopy confirmed the distribution of gadophrin-2 labeled cells to these target organs. Gadophrin-2 is suited as a bifunctional contrast agent for MR imaging, OI, and fluorescence microscopy and may be used to combine the advantages of each individual imaging modality for in vivo tracking of intravenously injected hematopoietic cells. (orig.)

  4. Development of Adaptive Tilt Tracker that Utilizes QUAD-cell Detector to Track Extended Objects

    2014-03-17

    tracked low Earth orbit (LEO) object and atmospheric seeing govern spot characteristics. Unlike static natural or laser guide stars, a LEO object’s...image spot characteristics .......................................................... 101 56. Response for non-adaptive tilt tracker with α equal to...applications toward natural and laser guide stars. The system was innovative and is a relevant forerunner to the tracker proposed in this research. The

  5. Monofunctional stealth nanoparticle for unbiased single molecule tracking inside living cells.

    Lisse, Domenik; Richter, Christian P; Drees, Christoph; Birkholz, Oliver; You, Changjiang; Rampazzo, Enrico; Piehler, Jacob

    2014-01-01

    On the basis of a protein cage scaffold, we have systematically explored intracellular application of nanoparticles for single molecule studies and discovered that recognition by the autophagy machinery plays a key role for rapid metabolism in the cytosol. Intracellular stealth nanoparticles were achieved by heavy surface PEGylation. By combination with a generic approach for nanoparticle monofunctionalization, efficient labeling of intracellular proteins with high fidelity was accomplished, allowing unbiased long-term tracking of proteins in the outer mitochondrial membrane.

  6. Track structure model for damage to mammalian cell cultures during solar proton events

    Cucinotta, F. A.; Wilson, J. W.; Townsend, L. W.; Shinn, J. L.; Katz, R.

    1992-01-01

    Solar proton events (SPEs) occur infrequently and unpredictably, thus representing a potential hazard to interplanetary space missions. Biological damage from SPEs will be produced principally through secondary electron production in tissue, including important contributions due to delta rays from nuclear reaction products. We review methods for estimating the biological effectiveness of SPEs using a high energy proton model and the parametric cellular track model. Results of the model are presented for several of the historically largest flares using typical levels and body shielding.

  7. Interacting active elastic dimers: Two cells moving on a rigid track

    Das, Moumita; Mayett, David; Schwarz, J. M.

    2015-03-01

    Cell migration in morphogenesis and cancer metastasis typically involves an interplay between different cell types. The rules governing such interplay remain largely unknown, however, a recent experiment studying the interaction between neural crest (NC) cells and placodal cells reveals an example of such rules. The study found that NC cells chase the placodal cells by chemotaxis, while placodal cells run away from NC cells when contacted by them. Motivated by this observation, we construct and study a minimal one-dimensional cell-cell model comprised of two cells with each cell represented by two-beads-connected-by-an-active spring. The active spring for each moving cell models the stress fibers with their myosin-driven contractility (and alpha-actinin extendability), while the friction coefficients of the beads describe the catch/slip bond behavior of the integrins in focal adhesions. We also include a dynamic contact interaction between the two cells, as well as a chemotactic potential, to decipher the chase-and-run dynamics observed in the experiment. We then use our modeling to further generalize the rules governing the interplay between different cell types during collective cell migration.

  8. Tracking Traction Force Changes of Single Cells on the Liquid Crystal Surface

    Chin Fhong Soon

    2015-01-01

    Full Text Available Cell migration is a key contributor to wound repair. This study presents findings indicating that the liquid crystal based cell traction force transducer (LCTFT system can be used in conjunction with a bespoke cell traction force mapping (CTFM software to monitor cell/surface traction forces from quiescent state in real time. In this study, time-lapse photo microscopy allowed cell induced deformations in liquid crystal coated substrates to be monitored and analyzed. The results indicated that the system could be used to monitor the generation of cell/surface forces in an initially quiescent cell, as it migrated over the culture substrate, via multiple points of contact between the cell and the surface. Future application of this system is the real-time assaying of the pharmacological effects of cytokines on the mechanics of cell migration.

  9. Quantitative tracking of tumor cells in phase-contrast microscopy exploiting halo artifact pattern

    Kang, Mi-Sun; Song, Soo-Min; Lee, Hana; Kim, Myoung-Hee

    2012-03-01

    Tumor cell morphology is closely related to its invasiveness characteristics and migratory behaviors. An invasive tumor cell has a highly irregular shape, whereas a spherical cell is non-metastatic. Thus, quantitative analysis of cell features is crucial to determine tumor malignancy or to test the efficacy of anticancer treatment. We use phase-contrast microscopy to analyze single cell morphology and to monitor its change because it enables observation of long-term activity of living cells without photobleaching and phototoxicity, which is common in other fluorescence-labeled microscopy. Despite this advantage, there are image-level drawbacks to phase-contrast microscopy, such as local light effect and contrast interference ring, among others. Thus, we first applied a local filter to compensate for non-uniform illumination. Then, we used intensity distribution information to detect the cell boundary. In phase-contrast microscopy images, the cell normally appears as a dark region surrounded by a bright halo. As the halo artifact around the cell body is minimal and has an asymmetric diffusion pattern, we calculated the cross-sectional plane that intersected the center of each cell and was orthogonal to the first principal axis. Then, we extracted the dark cell region by level set. However, a dense population of cultured cells still rendered single-cell analysis difficult. Finally, we measured roundness and size to classify tumor cells into malignant and benign groups. We validated segmentation accuracy by comparing our findings with manually obtained results.

  10. SuperSegger: robust image segmentation, analysis and lineage tracking of bacterial cells.

    Stylianidou, Stella; Brennan, Connor; Nissen, Silas B; Kuwada, Nathan J; Wiggins, Paul A

    2016-11-01

    Many quantitative cell biology questions require fast yet reliable automated image segmentation to identify and link cells from frame-to-frame, and characterize the cell morphology and fluorescence. We present SuperSegger, an automated MATLAB-based image processing package well-suited to quantitative analysis of high-throughput live-cell fluorescence microscopy of bacterial cells. SuperSegger incorporates machine-learning algorithms to optimize cellular boundaries and automated error resolution to reliably link cells from frame-to-frame. Unlike existing packages, it can reliably segment microcolonies with many cells, facilitating the analysis of cell-cycle dynamics in bacteria as well as cell-contact mediated phenomena. This package has a range of built-in capabilities for characterizing bacterial cells, including the identification of cell division events, mother, daughter and neighbouring cells, and computing statistics on cellular fluorescence, the location and intensity of fluorescent foci. SuperSegger provides a variety of postprocessing data visualization tools for single cell and population level analysis, such as histograms, kymographs, frame mosaics, movies and consensus images. Finally, we demonstrate the power of the package by analyzing lag phase growth with single cell resolution. © 2016 John Wiley & Sons Ltd.

  11. Magnetic resonance imaging tracking of human adipose derived stromal cells within three-dimensional scaffolds for bone tissue engineering

    C Lalande

    2011-04-01

    Full Text Available For bone tissue engineering, human Adipose Derived Stem Cells (hADSCs are proposed to be associated with a scaffold for promoting bone regeneration. After implantation, cellularised scaffolds require a non-invasive method for monitoring their fate in vivo. The purpose of this study was to use Magnetic Resonance Imaging (MRI-based tracking of these cells, labelled with magnetic agents for in vivo longitudinal assessment. hADSCs were isolated from adipose tissue and labelled with USPIO-rhodamine (Ultrasmall SuperParamagnetic Iron Oxide. USPIO internalisation, absence of toxicity towards hADSCs, and osteogenic differentiation of the labelled cells were evaluated in standard culture conditions. Labelled cells were then seeded within a 3D porous polysaccharide-based scaffold and imaged in vitro using fluorescence microscopy and MRI. Cellularised scaffolds were implanted subcutaneously in nude mice and MRI analyses were performed from 1 to 28 d after implantation. In vitro, no effect of USPIO labelling on cell viability and osteogenic differentiation was found. USPIO were efficiently internalised by hADSCs and generated a high T2* contrast. In vivo MRI revealed that hADSCs remain detectable until 28 d after implantation and could migrate from the scaffold and colonise the area around it. These data suggested that this scaffold might behave as a cell carrier capable of both holding a cell fraction and delivering cells to the site of implantation. In addition, the present findings evidenced that MRI is a reliable technique to validate cell-seeding procedures in 3D porous scaffolds, and to assess the fate of hADSCs transplanted in vivo.

  12. Simulations of DSB Yields and Radiation-induced Chromosomal Aberrations in Human Cells Based on the Stochastic Track Structure Induced by HZE Particles

    Ponomarev, Artem; Plante, Ianik; George, Kerry; Wu, Honglu

    2014-01-01

    The formation of double-strand breaks (DSBs) and chromosomal aberrations (CAs) is of great importance in radiation research and, specifically, in space applications. We are presenting a new particle track and DNA damage model, in which the particle stochastic track structure is combined with the random walk (RW) structure of chromosomes in a cell nucleus. The motivation for this effort stems from the fact that the model with the RW chromosomes, NASARTI (NASA radiation track image) previously relied on amorphous track structure, while the stochastic track structure model RITRACKS (Relativistic Ion Tracks) was focused on more microscopic targets than the entire genome. We have combined chromosomes simulated by RWs with stochastic track structure, which uses nanoscopic dose calculations performed with the Monte-Carlo simulation by RITRACKS in a voxelized space. The new simulations produce the number of DSBs as function of dose and particle fluence for high-energy particles, including iron, carbon and protons, using voxels of 20 nm dimension. The combined model also calculates yields of radiation-induced CAs and unrejoined chromosome breaks in normal and repair deficient cells. The joined computational model is calibrated using the relative frequencies and distributions of chromosomal aberrations reported in the literature. The model considers fractionated deposition of energy to approximate dose rates of the space flight environment. The joined model also predicts of the yields and sizes of translocations, dicentrics, rings, and more complex-type aberrations formed in the G0/G1 cell cycle phase during the first cell division after irradiation. We found that the main advantage of the joined model is our ability to simulate small doses: 0.05-0.5 Gy. At such low doses, the stochastic track structure proved to be indispensable, as the action of individual delta-rays becomes more important.

  13. Simulations of DSB Yields and Radiation-induced Chromosomal Aberrations in Human Cells Based on the Stochastic Track Structure iIduced by HZE Particles

    Ponomarev, Artem; Plante, Ianik; George, Kerry; Wu, Honglu

    2014-01-01

    The formation of double-strand breaks (DSBs) and chromosomal aberrations (CAs) is of great importance in radiation research and, specifically, in space applications. We are presenting a new particle track and DNA damage model, in which the particle stochastic track structure is combined with the random walk (RW) structure of chromosomes in a cell nucleus. The motivation for this effort stems from the fact that the model with the RW chromosomes, NASARTI (NASA radiation track image) previously relied on amorphous track structure, while the stochastic track structure model RITRACKS (Relativistic Ion Tracks) was focused on more microscopic targets than the entire genome. We have combined chromosomes simulated by RWs with stochastic track structure, which uses nanoscopic dose calculations performed with the Monte-Carlo simulation by RITRACKS in a voxelized space. The new simulations produce the number of DSBs as function of dose and particle fluence for high-energy particles, including iron, carbon and protons, using voxels of 20 nm dimension. The combined model also calculates yields of radiation-induced CAs and unrejoined chromosome breaks in normal and repair deficient cells. The joined computational model is calibrated using the relative frequencies and distributions of chromosomal aberrations reported in the literature. The model considers fractionated deposition of energy to approximate dose rates of the space flight environment. The joined model also predicts of the yields and sizes of translocations, dicentrics, rings, and more complex-type aberrations formed in the G0/G1 cell cycle phase during the first cell division after irradiation. We found that the main advantage of the joined model is our ability to simulate small doses: 0.05-0.5 Gy. At such low doses, the stochastic track structure proved to be indispensable, as the action of individual delta-rays becomes more important.

  14. Tracking neuronal marker expression inside living differentiating cells using molecular beacons

    Ilieva, Mirolyuba; Della Vedova, Paolo; Hansen, Ole

    2013-01-01

    and tyrosine hydroxylase mRNAs were expressed 2 and 3 days post induction of differentiation, respectively. Oct 4 was not detected with MB in these cells and signal was not increased over time suggesting that MB are generally stable inside the cells. The gene expression changes measured using MBs were...... confirmed using qRT-PCR. These results suggest that MBs are simple to use sensors inside living cell, and particularly useful for studying dynamic gene expression in heterogeneous cell populations....

  15. CD24 tracks divergent pluripotent states in mouse and human cells

    Shakiba, Nika; White, Carl A; Lipsitz, Yonatan Y; Yachie-Kinoshita, Ayako; Tonge, Peter D; Hussein, Samer M I; Puri, Mira C; Elbaz, Judith; Morrissey-Scoot, James; Li, Mira; Munoz Peralta, Javier; Benevento, Marco; Rogers, Ian M; Hanna, Jacob H; Heck, Albert J R; Wollscheid, Bernd; Nagy, Andras; Zandstra, Peter W

    2015-01-01

    Reprogramming is a dynamic process that can result in multiple pluripotent cell types emerging from divergent paths. Cell surface protein expression is a particularly desirable tool to categorize reprogramming and pluripotency as it enables robust quantification and enrichment of live cells. Here we

  16. Quantitative Label-Free Cell Proliferation Tracking with a Versatile Electrochemical Impedance Detection Platform

    Caviglia, Claudia; Carminati, M; Heiskanen, Arto

    2012-01-01

    optimal detection strategies. Electrochemical Impedance Spectroscopy (EIS) has been used to monitor and compare adhesion of different cell lines. HeLa cells and 3T3 fibroblasts have been cultured for 12 hours on interdigitated electrode arrays integrated into a tailor-made cell culture platform. Both......Since the use of impedance measurements for label-free monitoring of cells has become widespread but still the choice of sensing configuration is not unique though crucial for a quantitative interpretation of data, we demonstrate the application of a novel custom multipotentiostat platform to study...... vertical and coplanar interdigitated sensing configuration approaches have been used and compared on the same cell populations....

  17. Long-term MRI cell tracking after intraventricular delivery in a patient with global cerebral ischemia and prospects for magnetic navigation of stem cells within the CSF.

    Miroslaw Janowski

    Full Text Available The purpose of the study was to evaluate the long-term clinical tracking of magnetically labeled stem cells after intracerebroventricular transplantation as well as to investigate in vitro feasibility for magnetic guidance of cell therapy within large fluid compartments.After approval by our Institutional Review Board, an 18-month-old patient, diagnosed as being in a vegetative state due to global cerebral ischemia, underwent cell transplantation to the frontal horn of the lateral ventricle, with umbilical cord blood-derived stem cells labeled with superparamagnetic iron oxide (SPIO contrast agent. The patient was followed over 33 months with clinical examinations and MRI. To evaluate the forces governing the distribution of cells within the fluid compartment of the ventricular system in vivo, a gravity-driven sedimentation assay and a magnetic field-driven cell attraction assay were developed in vitro.Twenty-four hours post-transplantation, MR imaging (MRI was able to detect hypointense cells in the occipital horn of the lateral ventricle. The signal gradually decreased over 4 months and became undetectable at 33 months. In vitro, no significant difference in cell sedimentation between SPIO-labeled and unlabeled cells was observed (p = NS. An external magnet was effective in attracting cells over distances comparable to the size of human lateral ventricles.MR imaging of SPIO-labeled cells allows monitoring of cells within lateral ventricles. While the initial biodistribution is governed by gravity-driven sedimentation, an external magnetic field may possibly be applied to further direct the distribution of labeled cells within large fluid compartments such as the ventricular system.

  18. On the track of transfer cell formation by specialized plant-parasitic nematodes.

    Rodiuc, Natalia; Vieira, Paulo; Banora, Mohamed Youssef; de Almeida Engler, Janice

    2014-01-01

    Transfer cells are ubiquitous plant cells that play an important role in plant development as well as in responses to biotic and abiotic stresses. They are highly specialized and differentiated cells playing a central role in the acquisition, distribution and exchange of nutrients. Their unique structural traits are characterized by augmented ingrowths of invaginated secondary wall material, unsheathed by an amplified area of plasma membrane enriched in a suite of solute transporters. Similar morphological features can be perceived in vascular root feeding cells induced by sedentary plant-parasitic nematodes, such as root-knot and cyst nematodes, in a wide range of plant hosts. Despite their close phylogenetic relationship, these obligatory biotrophic plant pathogens engage different approaches when reprogramming root cells into giant cells or syncytia, respectively. Both nematode feeding-cells types will serve as the main source of nutrients until the end of the nematode life cycle. In both cases, these nematodes are able to remarkably maneuver and reprogram plant host cells. In this review we will discuss the structure, function and formation of these specialized multinucleate cells that act as nutrient transfer cells accumulating and synthesizing components needed for survival and successful offspring of plant-parasitic nematodes. Plant cells with transfer-like functions are also a renowned subject of interest involving still poorly understood molecular and cellular transport processes.

  19. On the track of transfer cells formation by specialized plant-parasitic nematodes

    Natalia eRodiuc

    2014-05-01

    Full Text Available Transfer cells are ubiquitous plant cells that play an important role in plant development as well as in responses to biotic and abiotic stresses. They are highly specialized and differentiated cells playing a central role in the acquisition, distribution and exchange of nutrients. Their unique structural traits are characterized by augmented ingrowths of invaginated secondary wall material, unsheathed by an amplified area of plasma membrane enriched in a suite of solute transporters. Similar morphological features can be perceived in vascular root feeding cells induced by sedentary plant-parasitic nematodes, such as root-knot and cyst nematodes, in a wide range of plant hosts. Despite their close phylogenetic relationship, these obligatory biotrophic plant pathogens engage different approaches when reprogramming root cells into giant cells or syncytia, respectively. Both nematode feeding-cells types will serve as the main source of nutrients until the end of the nematode life cycle. In both cases, these nematodes are able to remarkably maneuver and reprogram plant host cells. In this review we will discuss the structural, functional and morphogenetic characteristics function and formation of these specialized multinucleate cells that act as nutrient transfer cells to accumulate and synthesize components needed for survival and successful offspring of plant-parasitic nematodes. Plant cells with transfer-like functions are also a renowned subject of interest involving still poorly understood molecular and cellular transport processes.

  20. In vivo tracking of 111In-oxine labeled mesenchymal stem cells following infusion in patients with advanced cirrhosis

    Gholamrezanezhad, Ali; Mirpour, Sahar; Bagheri, Mohammad; Mohamadnejad, Mehdi; Alimoghaddam, Kamran; Abdolahzadeh, Leila; Saghari, Mohsen; Malekzadeh, Reza

    2011-01-01

    Background: Several animal and few human studies suggest the beneficial role of bone marrow mesenchymal stem cells (MSCs) in liver cirrhosis. However, little is known about the fate of MSCs after infusion in cirrhotic patients. We evaluated stem cell biodistribution after peripheral infusion of MSCs in four cirrhotic patients. Methods: After three passages of MSCs, the patients received a total of 250-400x10 6 cells, of which only 50% of the cells were labeled. Specific activities of 0.21-0.67 MBq/10 6 cells were maintained for the injected labeled MSCs. Planar whole-body acquisitions (anterior/posterior projections) were acquired immediately following infusion as well as at 2 h, 4 h, 6 h, 24 h, 48 h, 7th and 10th days after cell infusion. Results: After intravenous infusion, the radioactivity was first observed to accumulate in the lungs. During the following hours to days, the radioactivity gradually increased in the liver and spleen, with spleen uptake exceeding that in the liver in all patients. Region-of-interest analysis showed that the percentage of cells homing to the liver (following decay and background corrections and geometric mean calculation) increased from 0.0%-2.8% at immediately post-infusion images to 13.0-17.4% in 10th-day post-infusion. Similarly, the residual activities in the spleen increased from 2.0%-10.2% at immediately post-infusion images to 30.1%-42.2% in 10th-day post-infusion. During the same period, the residual activities in the lungs decreased from 27.0-33.5% to 2.0-5.4%. Conclusion: The infusion of MSCs labeled with 111 In-oxine through a peripheral vein is safe in cirrhosis. Cell labeling with 111 In-oxine is a suitable method for tracking MSC distribution after infusion.

  1. In vivo tracking of {sup 111}In-oxine labeled mesenchymal stem cells following infusion in patients with advanced cirrhosis

    Gholamrezanezhad, Ali, E-mail: agholam1@jhmi.edu [Research Institute for Nuclear Medicine. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Mirpour, Sahar [Research Institute for Nuclear Medicine. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Bagheri, Mohammad; Mohamadnejad, Mehdi [Digestive Disease Research Center. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Alimoghaddam, Kamran [Hematology and BMT Research Center. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Abdolahzadeh, Leila [Digestive Disease Research Center. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Saghari, Mohsen [Research Institute for Nuclear Medicine. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of); Malekzadeh, Reza [Digestive Disease Research Center. Shariati Hospital. Tehran University of Medical Sciences, Tehran, 14114 (Iran, Islamic Republic of)

    2011-10-15

    Background: Several animal and few human studies suggest the beneficial role of bone marrow mesenchymal stem cells (MSCs) in liver cirrhosis. However, little is known about the fate of MSCs after infusion in cirrhotic patients. We evaluated stem cell biodistribution after peripheral infusion of MSCs in four cirrhotic patients. Methods: After three passages of MSCs, the patients received a total of 250-400x10{sup 6} cells, of which only 50% of the cells were labeled. Specific activities of 0.21-0.67 MBq/10{sup 6} cells were maintained for the injected labeled MSCs. Planar whole-body acquisitions (anterior/posterior projections) were acquired immediately following infusion as well as at 2 h, 4 h, 6 h, 24 h, 48 h, 7th and 10th days after cell infusion. Results: After intravenous infusion, the radioactivity was first observed to accumulate in the lungs. During the following hours to days, the radioactivity gradually increased in the liver and spleen, with spleen uptake exceeding that in the liver in all patients. Region-of-interest analysis showed that the percentage of cells homing to the liver (following decay and background corrections and geometric mean calculation) increased from 0.0%-2.8% at immediately post-infusion images to 13.0-17.4% in 10th-day post-infusion. Similarly, the residual activities in the spleen increased from 2.0%-10.2% at immediately post-infusion images to 30.1%-42.2% in 10th-day post-infusion. During the same period, the residual activities in the lungs decreased from 27.0-33.5% to 2.0-5.4%. Conclusion: The infusion of MSCs labeled with {sup 111}In-oxine through a peripheral vein is safe in cirrhosis. Cell labeling with {sup 111}In-oxine is a suitable method for tracking MSC distribution after infusion.

  2. Magnetic resonance imaging with superparamagnetic iron oxide fails to track the long-term fate of mesenchymal stem cells transplanted into heart.

    Ma, Ning; Cheng, Huaibing; Lu, Minjie; Liu, Qiong; Chen, Xiuyu; Yin, Gang; Zhu, Hao; Zhang, Lianfeng; Meng, Xianmin; Tang, Yue; Zhao, Shihua

    2015-03-12

    MRI for in vivo stem cell tracking remains controversial. Here we tested the hypothesis that MRI can track the long-term fate of the superparamagnetic iron oxide (SPIO) nanoparticles labelled mesenchymal stem cells (MSCs) following intramyocardially injection in AMI rats. MSCs (1 × 10(6)) from male rats doubly labeled with SPIO and DAPI were injected 2 weeks after myocardial infarction. The control group received cell-free media injection. In vivo serial MRI was performed at 24 hours before cell delivery (baseline), 3 days, 1, 2, and 4 weeks after cell delivery, respectively. Serial follow-up MRI demonstrated large persistent intramyocardial signal-voids representing SPIO during the follow-up of 4 weeks, and MSCs did not moderate the left ventricular dysfunction. The TUNEL analysis confirmed that MSCs engrafted underwent apoptosis. The histopathological studies revealed that the site of cell injection was infiltrated by inflammatory cells progressively and the iron-positive cells were macrophages identified by CD68 staining, but very few or no DAPI-positive stem cells at 4 weeks after cells transplantation. The presence of engrafted cells was confirmed by real-time PCR, which showed that the amount of Y-chromosome-specific SRY gene was consistent with the results. MRI may not reliably track the long-term fate of SPIO-labeled MSCs engraftment in heart.

  3. Tracking of [{sup 18}F]FDG-labeled natural killer cells to HER2/neu-positive tumors

    Meier, Reinhard [Department of Radiology, University of California San Francisco (United States)], E-mail: reinhardt.meier@gmail.com; Piert, Morand [Department of Radiology, Division of Nuclear Medicine, University of Michigan (United States); Piontek, Guido; Rudelius, Martina [Institute of Pathology, Klinikum rechts der Isar, Technische Universitaet Muenchen (Germany); Oostendorp, Robert A. [3rd Department of Internal Medicine, Klinikum rechts der Isar, Technische Universitaet Muenchen (Germany); Senekowitsch-Schmidtke, Reingard [Department of Nuclear Medicine, Klinikum rechts der Isar, Technische Universitaet Muenchen (Germany); Henning, Tobias D. [Department of Radiology, University of California San Francisco (United States); Wels, Winfried S.; Uherek, Christoph [Chemotherapeutisches Forschungsinstitut, Georg-Speyer-Haus, Frankfurt am Main (Germany); Rummeny, Ernst J. [Department of Radiology, Klinikum rechts der Isar, Technische Universitaet Muenchen (Germany); Daldrup-Link, Heike E. [Department of Radiology, University of California San Francisco (United States)

    2008-07-15

    Introduction: The objective of this study was to label the human natural killer (NK) cell line NK-92 with [{sup 18}F]fluoro-deoxy-glucose (FDG) for subsequent in vivo tracking to HER2/neu-positive tumors. Methods: NK-92 cells were genetically modified to NK-92-scFv(FRP5)-zeta cells, which express a chimeric antigen receptor that is specific to the tumor-associated ErbB2 (HER2/neu) antigen. NK-92 and NK-92-scFv(FRP5)-zeta cells were labeled with [{sup 18}F]FDG by simple incubation at different settings. Labeling efficiency was evaluated by a gamma counter. Subsequently, [{sup 18}F]FDG-labeled parental NK-92 or NK-92-scFv(FRP5)-zeta cells were intravenously injected into mice with implanted HER2/neu-positive NIH/3T3 tumors. Radioactivity in tumors was quantified by digital autoradiography and correlated with histopathology. Results: The NK-92 and NK-92-scFv(FRP5)-zeta cells could be efficiently labeled with [{sup 18}F]FDG by simple incubation. Optimal labeling efficiencies (80%) were achieved using an incubation period of 60 min and additional insulin (10 IU/ml). After injection of 5x10{sup 6} [{sup 18}F]FDG-labeled NK-92-scFv(FRP5)-zeta cells into tumor-bearing mice, digital autoradiography showed an increased uptake of radioactivity in HER2/neu-positive tumors at 60 min postinjection. Conversely, injection of 5x10{sup 6} NK-92 cells not directed against HER2/neu receptors did not result in increased uptake of radioactivity in the tumors. Histopathology confirmed an accumulation of the NK-92-scFv(FRP5)-zeta cells, but not the parental NK cells, in tumor tissues. Conclusion: The human NK cell line NK-92 can be directed against HER2/neu antigens by genetic modification. The genetically modified NK cells can be efficiently labeled with [{sup 18}F]FDG, and the accumulation of these labeled NK cells in HER2/neu-positive tumors can be monitored with autoradiography.

  4. Tracking of [18F]FDG-labeled natural killer cells to HER2/neu-positive tumors

    Meier, Reinhard; Piert, Morand; Piontek, Guido; Rudelius, Martina; Oostendorp, Robert A.; Senekowitsch-Schmidtke, Reingard; Henning, Tobias D.; Wels, Winfried S.; Uherek, Christoph; Rummeny, Ernst J.; Daldrup-Link, Heike E.

    2008-01-01

    Introduction: The objective of this study was to label the human natural killer (NK) cell line NK-92 with [ 18 F]fluoro-deoxy-glucose (FDG) for subsequent in vivo tracking to HER2/neu-positive tumors. Methods: NK-92 cells were genetically modified to NK-92-scFv(FRP5)-zeta cells, which express a chimeric antigen receptor that is specific to the tumor-associated ErbB2 (HER2/neu) antigen. NK-92 and NK-92-scFv(FRP5)-zeta cells were labeled with [ 18 F]FDG by simple incubation at different settings. Labeling efficiency was evaluated by a gamma counter. Subsequently, [ 18 F]FDG-labeled parental NK-92 or NK-92-scFv(FRP5)-zeta cells were intravenously injected into mice with implanted HER2/neu-positive NIH/3T3 tumors. Radioactivity in tumors was quantified by digital autoradiography and correlated with histopathology. Results: The NK-92 and NK-92-scFv(FRP5)-zeta cells could be efficiently labeled with [ 18 F]FDG by simple incubation. Optimal labeling efficiencies (80%) were achieved using an incubation period of 60 min and additional insulin (10 IU/ml). After injection of 5x10 6 [ 18 F]FDG-labeled NK-92-scFv(FRP5)-zeta cells into tumor-bearing mice, digital autoradiography showed an increased uptake of radioactivity in HER2/neu-positive tumors at 60 min postinjection. Conversely, injection of 5x10 6 NK-92 cells not directed against HER2/neu receptors did not result in increased uptake of radioactivity in the tumors. Histopathology confirmed an accumulation of the NK-92-scFv(FRP5)-zeta cells, but not the parental NK cells, in tumor tissues. Conclusion: The human NK cell line NK-92 can be directed against HER2/neu antigens by genetic modification. The genetically modified NK cells can be efficiently labeled with [ 18 F]FDG, and the accumulation of these labeled NK cells in HER2/neu-positive tumors can be monitored with autoradiography

  5. Molecular beacon nanosensors for live cell detection and tracking differentiation and reprogramming

    Ilieva, Mirolyuba

    2013-01-01

    open to closed state within living cells. Using MBs targeting pluripotent stem cell markers we demonstrated reverse into a more immature state of LUHMES induced by neurosphere-like growth conditions. Moreover, we have been able to trace localisation of this particular population during differentiation...... in separation of fluorophore from quencher and thereby emission of a fluorescent signal that can be detected. In this project the usability and applicability of MBs for live cell detection and tracing of gene expression was demonstrated. MBs library targeting gene markers for pluripotent stem cells as well...... and thus demonstrate the usability of MBs for monitoring cell behaviour within 3D clusters. Finally, MBs detection of expression of human pluripotent markers after reprograming of adult somatic cells with plasmid codding for mouse transcription factors was demonstrated. In conclusion, the method of using...

  6. Detecting carbon uptake and cellular allocation by individual algae in multispecies assemblages: Tracking carbon into single algal cells

    Murdock, Justin N. [USDA Agricultural Research Service, National Sedimentation Laboratory, Oxford Mississippi; Department of Biology, Tennessee Technological University, Cookeville Tennessee

    2015-11-03

    Algal species vary in carbon (C) need and uptake rates. Understanding differences in C uptake and cellular allocation among species from natural communities will bring new insight into many ecosystem process questions including how species changes will alter energy availability and C sequestration in aquatic ecosystems. A major limitation of current methods that measure algal C incorporation is the inability to separate the response of individual species from mixed-species assemblages. I used Fourier-transform infrared microspectroscopy to qualitatively measure inorganic 13C isotope incorporation into individual algal cells in single species, two species, and natural phytoplankton assemblages. Lateral shifts in spectral peaks from 13C treatments were observed in all species. Comparison of peaks associated with carbohydrates, proteins, and lipids allowed for the detection of which individuals took in C, and which macromolecules the C was used to make. For example, shifts in Spirogyra spectral peaks showed substantial C incorporation in carbohydrates. Further, shifts in peaks at 1160 cm-1, 1108 cm-1, 1080 cm-1, 1048 cm-1, and 1030 cm-1 suggested C was being allocated into cellulose. The natural phytoplankton assemblage demonstrated how C could be tracked into co-occurring species. A diatom had large shifts in protein and carbohydrate peaks, while a green alga and euglenoid had only a few shifts in protein related peaks. Fourier-transform infrared microspectroscopy is an established, label free method for measuring the chemical composition of algal cells. However, adding a label such as 13C isotope can greatly expand the technique's capabilities by qualitatively tracking C movement between inorganic and organic states within single cells.

  7. Development and assessment of transparent soil and particle image velocimetry in dynamic soil-structure interaction

    2007-02-01

    This research combines Particle Image Velocimetry (PIV) and transparent soil to investigate the dynamic rigid block and soil interaction. In order to get a low viscosity pore fluid for the transparent soil, 12 different types of chemical solvents wer...

  8. 3D Flow Field Measurements using Aerosol Correlation Velocimetry, Phase II

    National Aeronautics and Space Administration — AeroMancer Technologies proposes to develop a 3D Global Lidar Airspeed Sensor (3D-LGAS) using Aerosol Correlation Velocimetry for standoff sensing of high-resolution...

  9. Demonstration of Clean Particle Seeding for Particle Image Velocimetry in a Closed Circuit Supersonic Wind Tunnel

    McNiel, Charles M

    2007-01-01

    The purpose of this research was to determine whether solid carbon dioxide (CO2) particles might provide a satisfactory, and cleaner, alternative to traditional seed material for Particle Image Velocimetry (PIV...

  10. Molecular Imaging of Stem Cells: Tracking Survival, Biodistribution, Tumorigenicity, and Immunogenicity

    Eugene Gu, Wen-Yi Chen, Jay Gu, Paul Burridge, Joseph C. Wu

    2012-01-01

    Full Text Available Being able to self-renew and differentiate into virtually all cell types, both human embryonic stem cells (hESCs and induced pluripotent stem cells (iPSCs have exciting therapeutic implications for myocardial infarction, neurodegenerative disease, diabetes, and other disorders involving irreversible cell loss. However, stem cell biology remains incompletely understood despite significant advances in the field. Inefficient stem cell differentiation, difficulty in verifying successful delivery to the target organ, and problems with engraftment all hamper the transition from laboratory animal studies to human clinical trials. Although traditional histopathological techniques have been the primary approach for ex vivo analysis of stem cell behavior, these postmortem examinations are unable to further elucidate the underlying mechanisms in real time and in vivo. Fortunately, the advent of molecular imaging has led to unprecedented progress in understanding the fundamental behavior of stem cells, including their survival, biodistribution, immunogenicity, and tumorigenicity in the targeted tissues of interest. This review summarizes various molecular imaging technologies and how they have advanced the current understanding of stem cell survival, biodistribution, immunogenicity, and tumorigenicity.

  11. In vivo tracking of magnetically labeled mesenchmal stem cells injected via renal arteries in kidney failure rat

    Sun Junhui; Teng Gaojun; Ju Shenghong; Ma Zhanlong; Mai Xiaoli; Zhang Yu; Ma Ming

    2006-01-01

    Objective: To evaluate in vivo depiction and tracking for magnetically labeled bone marrow mesenchymal stern cells (MSCs) in a renal failure rat model injected intravascularly using a 1.5 T magnetic resonance imaging (MRI) system. Methods: Rat MSCs were isolated, purified, expanded and then incubated with home synthesized Fe 2 O 3 -PLL. Prussian blue stain was employed for identifying intracellular irons. An acute renal failure in rat was induced by intramuscular injection of glycerol and MSCs were injected into renal arteries of 11 recipients (labeled cells in six, unlabeled cells in five). MR images of kidneys were obtained respectively before injection of MSCs, and immediately, 1, 3, 5, and 8 clays after transplantation. MR imaging findings were analyzed, which were correlated with histological findings. Results: Rat MSCs were successfully labeled, and labeling efficiency was almost 100%. Prussian blue staining of Fe 2 O 3 -PLL labeled cells revealed the presence of iron-containing vesicles or endosomes in the cytoplasm. In the renal failure model of rats, the labeled MSCs were demonstrated as signal intensity loss in renal cortex on T 2 * -weighted MR images. The signal intensity decrease was visualized up to days 8 after transplantation. Histological analyses showed that most Prussian blue staining-positive cells were well correlated with the area where a signal intensity loss was observed in MRI. Signal intensity decrease was not detected after transplantation of unlabeled cells. Conclusion: The rat MSCs can be effectively labeled with Fe 2 O 3 -PLL. 1.5-T MR imaging seems to be a good technique to monitor the magnetically labeled MSCs in vivo in renal failure rat model intravascularly administered, which may have much more potential values for studying the engraftment of stem cells in kidneys. (authors)

  12. In vivo human adipose-derived mesenchymal stem cell tracking after intra-articular delivery in a rat osteoarthritis model

    Meng Li

    2016-11-01

    Full Text Available Abstract Background Human adipose-derived mesenchymal stem cells (haMSCs have shown efficacy in treating osteoarthritis (OA both preclinically and clinically via intra-articular (IA injection. However, understanding the mode of action of the cell therapy has been limited by cell tracking capability and correlation between the pharmacokinetics of the injected cells and the intended pharmacodynamics effect. This study aims to explore methodology and to understand in vivo biodistribution of clinical-grade haMSCs labeled with fluorescent dye and injected into an immunocompetent OA rat model. Methods haMSCs labeled with fluorescent dye were investigated for their proliferation and differentiation capabilities. Labeled cells were used to establish detection threshold of a noninvasive biofluorescent imaging system before the cells (2.5 × 106 were injected into a conventional rat OA model induced by medial meniscectomy for 8 weeks. We attempted to reveal the existence of labeled cells in vivo by imaging and a molecular biomarker approach, and to correlate with the in vivo efficacy and physical presence over a follow-up period up to 10 weeks. Results In vitro proliferation and differentiation of haMSCs were not affected by the labeling of DiD dye. Detection thresholds of the labeled cells in vitro and in vivo were determined to be 104 and 105 cells, respectively. When 2.5 × 106 haMSCs were injected into the joints of a rat OA model, fluorescent signals (or >105 cells lasted for about 10 weeks in the surgical knee joint at the same time as efficacy was observed. Signals in nonsurgical rats only lasted for 4 weeks. The human MSCs were shown to engraft to the rat joint tissues and were proliferative. Human FOXP2 gene was only detected in the knee joint tissue, suggesting limited biodistribution locally to the joints. Conclusions The current study represents the first attempt to correlate cell therapy efficacy on OA with the physical presence

  13. Estimating Horizontal Displacement between DEMs by Means of Particle Image Velocimetry Techniques

    Juan F. Reinoso

    2015-12-01

    Full Text Available To date, digital terrain model (DTM accuracy has been studied almost exclusively by computing its height variable. However, the largely ignored horizontal component bears a great influence on the positional accuracy of certain linear features, e.g., in hydrological features. In an effort to fill this gap, we propose a means of measurement different from the geomatic approach, involving fluid mechanics (water and air flows or aerodynamics. The particle image velocimetry (PIV algorithm is proposed as an estimator of horizontal differences between digital elevation models (DEM in grid format. After applying a scale factor to the displacement estimated by the PIV algorithm, the mean error predicted is around one-seventh of the cell size of the DEM with the greatest spatial resolution, and around one-nineteenth of the cell size of the DEM with the least spatial resolution. Our methodology allows all kinds of DTMs to be compared once they are transformed into DEM format, while also allowing comparison of data from diverse capture methods, i.e., LiDAR versus photogrammetric data sources.

  14. Tracking Solid Oxide Cell Microstructure Evolution by High Resolution 3D Nano-Tomography

    De Angelis, Salvatore

    . The degradation processes are mainly attributed to morphological changes occurring within the electrodes microstructure. Therefore, precise tracking of 3D microstructural evolution during operation is considered crucial to understanding the complex relationship between microstructure and performance. In this work......, X-ray ptychographic tomography is applied to SOC materials, demonstrating unprecedented spatial resolution and data quality. The eect of a complete redox cycle on the same Ni-YSZ microstructure is visualized ex-situ in 3D, showing major rearrangement of the nickel network after reduction......, the formation of cracks in the YSZ, and void formation in nickel oxide after oxidation. Capitalizing on the high resolution of ptychography, the eect of nickel coarsening on the Ni-YSZ microstructure evolution is studied ex-situ in three dimensions, while the sample is repeatedly scanned and treated at high...

  15. Stereotactic radiotherapy with real-time tumor tracking for non-small cell lung cancer: Clinical outcome

    Voort van Zyp, Noelle C. van der; Prevost, Jean-Briac; Hoogeman, Mischa S.; Praag, John; Holt, Bronno van der; Levendag, Peter C.; Klaveren, Robertus J. van; Pattynama, Peter; Nuyttens, Joost J.

    2009-01-01

    Purpose: To report the clinical outcome of treatment using real-time tumor tracking for 70 patients with inoperable stage I non-small cell lung cancer (NSCLC). Materials and methods: Seventy inoperable patients with peripherally located early-stage NSCLC were treated with 45 or 60 Gy in three fractions using CyberKnife. Pathology was available in 51% of patients. Thirty-nine patients had a T1-tumor and 31 had a T2-tumor. Markers were placed using the vascular, percutaneous intra-, or extra-pulmonary approach, depending on the risk of pneumothorax. Results: The actuarial 2-year local control rate for patients treated with 60 Gy was 96%, compared to 78% for patients treated with a total dose of 45 Gy (p = 0.197). All local recurrences (n = 4) occurred in patients with T2-tumors. Overall survival for the whole group at two years was 62% and the cause specific survival was 85%. The median follow-up was 15 months. Grade 3 toxicity occurred in two patients (3%) after marker placement. Treatment-related late grade 3 toxicity occurred in 7 patients (10%). No grade ≥4 toxicity occurred. Conclusion: Excellent local control of 96% at 1- and 2-years was achieved using 60 Gy in three fractions for NSCLC patients treated with the real-time tumor tracking. Toxicity was low.

  16. Homing and Tracking of Iron Oxide Labelled Mesenchymal Stem Cells After Infusion in Traumatic Brain Injury Mice: a Longitudinal In Vivo MRI Study.

    Mishra, Sushanta Kumar; Khushu, Subash; Singh, Ajay K; Gangenahalli, Gurudutta

    2018-06-17

    Stem cells transplantation has emerged as a promising alternative therapeutic due to its potency at injury site. The need to monitor and non-invasively track the infused stem cells is a significant challenge in the development of regenerative medicine. Thus, in vivo tracking to monitor infused stem cells is especially vital. In this manuscript, we have described an effective in vitro labelling method of MSCs, a serial in vivo tracking of implanted stem cells at traumatic brain injury (TBI) site through 7 T magnetic resonance imaging (MRI). Proper homing of infused MSCs was carried out at different time points using histological analysis and Prussian blue staining. Longitudinal in vivo tracking of infused MSCs were performed up to 21 days in different groups through MRI using relaxometry technique. Results demonstrated that MSCs incubated with iron oxide-poly-L-lysine complex (IO-PLL) at a ratio of 50:1.5 μg/ml and a time period of 6 h was optimised to increase labelling efficiency. T2*-weighted images and relaxation study demonstrated a significant signal loss and effective decrease in transverse relaxation time on day-3 at injury site after systemic transplantation, revealed maximum number of stem cells homing to the lesion area. MRI results further correlate with histological and Prussian blue staining in different time periods. Decrease in negative signal and increase in relaxation times were observed after day-14, may indicate damage tissue replacement with healthy tissue. MSCs tracking with synthesized negative contrast agent represent a great advantage during both in vitro and in vivo analysis. The proposed absolute bias correction based relaxometry analysis could be extrapolated for stem cell tracking and therapies in various neurodegenerative diseases.

  17. New robust algorithm for tracking cells in videos of Drosophila morphogenesis based on finding an ideal path in segmented spatio-temporal cellular structures.

    Bellaïche, Yohanns; Bosveld, Floris; Graner, François; Mikula, Karol; Remesíková, Mariana; Smísek, Michal

    2011-01-01

    In this paper, we present a novel algorithm for tracking cells in time lapse confocal microscopy movie of a Drosophila epithelial tissue during pupal morphogenesis. We consider a 2D + time video as a 3D static image, where frames are stacked atop each other, and using a spatio-temporal segmentation algorithm we obtain information about spatio-temporal 3D tubes representing evolutions of cells. The main idea for tracking is the usage of two distance functions--first one from the cells in the initial frame and second one from segmented boundaries. We track the cells backwards in time. The first distance function attracts the subsequently constructed cell trajectories to the cells in the initial frame and the second one forces them to be close to centerlines of the segmented tubular structures. This makes our tracking algorithm robust against noise and missing spatio-temporal boundaries. This approach can be generalized to a 3D + time video analysis, where spatio-temporal tubes are 4D objects.

  18. Analysis of bubbly flow using particle image velocimetry

    Todd, D.R.; Ortiz-Villafuerte, J.; Schmidl, W.D.; Hassan, Y.A. [Texas A and M University, Nuclear Engineering Dept., College Stagion, TX (United States); Sanchez-Silva, F. [ESIME, INP (Mexico)

    2001-07-01

    The local phasic velocities can be determined in two-phase flows if the phases can be separated during analysis. The continuous liquid velocity field can be captured using standard Particle Image Velocimetry (PIV) techniques in two-phase flows. PIV is now a well-established, standard flow measurement technique, which provides instantaneous velocity fields in a two-dimensional plane of finite thickness. PIV can be extended to three dimensions within the plane with special considerations. A three-dimensional shadow PIV (SPIV) measurement apparatus can be used to capture the dispersed phase flow parameters such as velocity and interfacial area. The SPIV images contain only the bubble images, and can be easily analyzed and the results used to separate the dispersed phase from the continuous phase in PIV data. An experimental system that combines the traditional PIV technique with SPIV will be described and sample data will be analyzed to demonstrate an advanced turbulence measurement method in a two-phase bubbly flow system. Also, a qualitative error analysis method that allows users to reduce the number of erroneous vectors obtained from the PIV measurements will be discussed. (authors)

  19. Particle Image Velocimetry (PIV) Measurements of Suspension-Feeding Velocities

    Du Clos, K.; Jones, I. T.; Carrier, T. J.; Jumars, P. A.

    2016-02-01

    Active suspension feeders, such as bivalves and tunicates, connect benthic and pelagic ecosystems by packaging suspended matter into larger fecal and pseudofecal particles, greatly enhancing the flux of carbon and nutrients from the water column to the benthos. The volume of water processed by a population of suspension feeders is commonly estimated by scaling up results from experiments that measure the clearance rate (the volume of water cleared of particles per time) of one or a few individual suspension feeders. Clearance rates vary, however, between species, within a species, and over time for a single individual; and the velocity fields produced by suspension feeders are likely to interact in complex ways. We measured the water velocity fields produced by two species of bivalve, Mya arenaria and Mercenaria mercenaria, and the tunicate Ciona intestinalis, using particle image velocimetry (PIV). We used these measurements to calculate flow rates and Reynolds numbers of inhalant and exhalant siphons. We also observed strong entrainment of water by M. arenaria's exhalant siphon jet that may help to explain how the clam avoids depleting the water around it of particles and oxygen as it feeds. We are using these measurements to inform computational fluid mechanics (CFD) models of suspension feeding, allowing us to examine the interactions of flow fields produced by multiple suspension feeders and other effects not quantified by clearance-rate measurements.

  20. Evaluation of magnetic resonance velocimetry for steady flow.

    Ku, D N; Biancheri, C L; Pettigrew, R I; Peifer, J W; Markou, C P; Engels, H

    1990-11-01

    Whole body magnetic resonance (MR) imaging has recently become an important diagnostic tool for cardiovascular diseases. The technique of magnetic resonance phase velocity encoding allows the quantitative measurement of velocity for an arbitrary component direction. A study was initiated to determine the ability and accuracy of MR velocimetry to measure a wide range of flow conditions including flow separation, three-dimensional secondary flow, high velocity gradients, and turbulence. A steady flow system pumped water doped with manganese chloride through a variety of test sections. Images were produced using gradient echo sequences on test sections including a straight tube, a curved tube, a smoothly converging-diverging nozzle, and an orifice. Magnetic resonance measurements of laminar and turbulent flows were depicted as cross-sectional velocity profiles. MR velocity measurements revealed such flow behavior as spatially varying velocity, recirculation and secondary flows over a wide range of conditions. Comparisons made with published experimental laser Doppler anemometry measurements and theoretical calculations for similar flow conditions revealed excellent accuracy and precision levels. The successful measurement of velocity profiles for a variety of flow conditions and geometries indicate that magnetic resonance imaging is an accurate, non-contacting velocimeter.

  1. Application of adaptive Kalman filter in vehicle laser Doppler velocimetry

    Fan, Zhe; Sun, Qiao; Du, Lei; Bai, Jie; Liu, Jingyun

    2018-03-01

    Due to the variation of road conditions and motor characteristics of vehicle, great root-mean-square (rms) error and outliers would be caused. Application of Kalman filter in laser Doppler velocimetry(LDV) is important to improve the velocity measurement accuracy. In this paper, the state-space model is built by using current statistical model. A strategy containing two steps is adopted to make the filter adaptive and robust. First, the acceleration variance is adaptively adjusted by using the difference of predictive observation and measured observation. Second, the outliers would be identified and the measured noise variance would be adjusted according to the orthogonal property of innovation to reduce the impaction of outliers. The laboratory rotating table experiments show that adaptive Kalman filter greatly reduces the rms error from 0.59 cm/s to 0.22 cm/s and has eliminated all the outliers. Road experiments compared with a microwave radar show that the rms error of LDV is 0.0218 m/s, and it proves that the adaptive Kalman filtering is suitable for vehicle speed signal processing.

  2. Analysis of bubbly flow using particle image velocimetry

    Todd, D.R.; Ortiz-Villafuerte, J.; Schmidl, W.D.; Hassan, Y.A.; Sanchez-Silva, F.

    2001-01-01

    The local phasic velocities can be determined in two-phase flows if the phases can be separated during analysis. The continuous liquid velocity field can be captured using standard Particle Image Velocimetry (PIV) techniques in two-phase flows. PIV is now a well-established, standard flow measurement technique, which provides instantaneous velocity fields in a two-dimensional plane of finite thickness. PIV can be extended to three dimensions within the plane with special considerations. A three-dimensional shadow PIV (SPIV) measurement apparatus can be used to capture the dispersed phase flow parameters such as velocity and interfacial area. The SPIV images contain only the bubble images, and can be easily analyzed and the results used to separate the dispersed phase from the continuous phase in PIV data. An experimental system that combines the traditional PIV technique with SPIV will be described and sample data will be analyzed to demonstrate an advanced turbulence measurement method in a two-phase bubbly flow system. Also, a qualitative error analysis method that allows users to reduce the number of erroneous vectors obtained from the PIV measurements will be discussed. (authors)

  3. Particle image velocimetry investigation of a finite amplitude pressure wave

    Thornhill, D.; Currie, T.; Fleck, R.; Chatfield, G.

    2006-03-01

    Particle image velocimetry is used to study the motion of gas within a duct subject to the passage of a finite amplitude pressure wave. The wave is representative of the pressure waves found in the exhaust systems of internal combustion engines. Gas particles are accelerated from stationary to 150 m/s and then back to stationary in 8 ms. It is demonstrated that gas particles at the head of the wave travel at the same velocity across the duct cross section at a given point in time. Towards the tail of the wave viscous effects are plainly evident causing the flow profile to tend towards parabolic. However, the instantaneous mean particle velocity across the section is shown to match well with the velocity calculated from a corresponding measured pressure history using 1D gas dynamic theory. The measured pressure history at a point in the duct was acquired using a high speed pressure transducer of the type typically used for engine research in intake and exhaust systems. It is demonstrated that these are unable to follow the rapid changes in pressure accurately and that they are prone to resonate under certain circumstances.

  4. Channel flow structure measurements using particle image velocimetry

    Norazizi Mohamed; Noraeini Mokhtar; Aziz Ibrahim; Ramli Abu Hassan

    1996-01-01

    Two different flow structures in a laboratory channel were examined using a flow visualization technique, known as Particle Image Velocimetry (PIV). The first channel flow structure was that of a steady flow over a horizontal channel bottom. Photographs of particle displacements were taken in the boundary layer in a plane parallel to the flow. These photographs were analyzed to give simultaneous measurements of two components of the velocity at hundreds of points in the plane. Averaging these photographs gave the velocity profile a few millimeters from the bottom of the channel to the water surface. The results gave good agreement with the known boundary layer theory. This technique is extended to the study of the structure under a progressive wave in the channel. A wavelength of the propagating wave is divided into sections by photographing it continously for a number of frames. Each frame is analyzed and a velocity field under this wave at various phase points were produced with their respective directions. The results show that velocity vectors in a plane under the wave could be achieved instantaneously and in good agreement with the small amplitude wave theory

  5. Zebrafish swimming in the flow: a particle image velocimetry study

    Violet Mwaffo

    2017-11-01

    Full Text Available Zebrafish is emerging as a species of choice for the study of a number of biomechanics problems, including balance development, schooling, and neuromuscular transmission. The precise quantification of the flow physics around swimming zebrafish is critical toward a mechanistic understanding of the complex swimming style of this fresh-water species. Although previous studies have elucidated the vortical structures in the wake of zebrafish swimming in placid water, the flow physics of zebrafish swimming against a water current remains unexplored. In an effort to illuminate zebrafish swimming in a dynamic environment reminiscent of its natural habitat, we experimentally investigated the locomotion and hydrodynamics of a single zebrafish swimming in a miniature water tunnel using particle image velocimetry. Our results on zebrafish locomotion detail the role of flow speed on tail beat undulations, heading direction, and swimming speed. Our findings on zebrafish hydrodynamics offer a precise quantification of vortex shedding during zebrafish swimming and demonstrate that locomotory patterns play a central role on the flow physics. This knowledge may help clarify the evolutionary advantage of burst and cruise swimming movements in zebrafish.

  6. Displacement fields from point cloud data: Application of particle imaging velocimetry to landslide geodesy

    Aryal, Arjun; Brooks, Benjamin A.; Reid, Mark E.; Bawden, Gerald W.; Pawlak, Geno

    2012-01-01

    Acquiring spatially continuous ground-surface displacement fields from Terrestrial Laser Scanners (TLS) will allow better understanding of the physical processes governing landslide motion at detailed spatial and temporal scales. Problems arise, however, when estimating continuous displacement fields from TLS point-clouds because reflecting points from sequential scans of moving ground are not defined uniquely, thus repeat TLS surveys typically do not track individual reflectors. Here, we implemented the cross-correlation-based Particle Image Velocimetry (PIV) method to derive a surface deformation field using TLS point-cloud data. We estimated associated errors using the shape of the cross-correlation function and tested the method's performance with synthetic displacements applied to a TLS point cloud. We applied the method to the toe of the episodically active Cleveland Corral Landslide in northern California using TLS data acquired in June 2005–January 2007 and January–May 2010. Estimated displacements ranged from decimeters to several meters and they agreed well with independent measurements at better than 9% root mean squared (RMS) error. For each of the time periods, the method provided a smooth, nearly continuous displacement field that coincides with independently mapped boundaries of the slide and permits further kinematic and mechanical inference. For the 2010 data set, for instance, the PIV-derived displacement field identified a diffuse zone of displacement that preceded by over a month the development of a new lateral shear zone. Additionally, the upslope and downslope displacement gradients delineated by the dense PIV field elucidated the non-rigid behavior of the slide.

  7. A three-dimensional strain measurement method in elastic transparent materials using tomographic particle image velocimetry.

    Azuma Takahashi

    Full Text Available The mechanical interaction between blood vessels and medical devices can induce strains in these vessels. Measuring and understanding these strains is necessary to identify the causes of vascular complications. This study develops a method to measure the three-dimensional (3D distribution of strain using tomographic particle image velocimetry (Tomo-PIV and compares the measurement accuracy with the gauge strain in tensile tests.The test system for measuring 3D strain distribution consists of two cameras, a laser, a universal testing machine, an acrylic chamber with a glycerol water solution for adjusting the refractive index with the silicone, and dumbbell-shaped specimens mixed with fluorescent tracer particles. 3D images of the particles were reconstructed from 2D images using a multiplicative algebraic reconstruction technique (MART and motion tracking enhancement. Distributions of the 3D displacements were calculated using a digital volume correlation. To evaluate the accuracy of the measurement method in terms of particle density and interrogation voxel size, the gauge strain and one of the two cameras for Tomo-PIV were used as a video-extensometer in the tensile test. The results show that the optimal particle density and interrogation voxel size are 0.014 particles per pixel and 40 × 40 × 40 voxels with a 75% overlap. The maximum measurement error was maintained at less than 2.5% in the 4-mm-wide region of the specimen.We successfully developed a method to experimentally measure 3D strain distribution in an elastic silicone material using Tomo-PIV and fluorescent particles. To the best of our knowledge, this is the first report that applies Tomo-PIV to investigate 3D strain measurements in elastic materials with large deformation and validates the measurement accuracy.

  8. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    Seo, Ji Hye; Joo, Sang-Woo [Department of Chemistry, Soongsil University, Seoul 156-743 (Korea, Republic of); Cho, Keunchang [Logos Biosystems, Incorporated, Anyang 431-070 (Korea, Republic of); Lee, So Yeong, E-mail: leeso@snu.ac.kr, E-mail: sjoo@ssu.ac.kr [Laboratory of Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742 (Korea, Republic of)

    2011-06-10

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  9. Concentration-dependent fluorescence live-cell imaging and tracking of intracellular nanoparticles

    Seo, Ji Hye; Joo, Sang-Woo; Cho, Keunchang; Lee, So Yeong

    2011-01-01

    Using live-cell imaging techniques we investigated concentration-dependent intracellular movements of fluorescence nanoparticles (NPs) in real-time after their entry into HeLa cells via incubation. Intracellular particle traces appeared to be a mixture of both random and fairly unidirectional movements of the particles. At rather low concentrations of NPs, a majority of the non-random intracellular particle trajectories are assumed to mostly go along microtubule networks after endocytosis, as evidenced from the inhibition test with nocodazole. On the other hand, as the concentrations of NPs increased, random motions were more frequently observed inside the cells.

  10. Hierarchical Load Tracking Control of a Grid-connected Solid Oxide Fuel Cell for Maximum Electrical Efficiency Operation

    Li, Yonghui; Wu, Qiuwei; Zhu, Haiyu

    2015-01-01

    efficiency operation obtained at different active power output levels, a hierarchical load tracking control scheme for the grid-connected SOFC was proposed to realize the maximum electrical efficiency operation with the stack temperature bounded. The hierarchical control scheme consists of a fast active...... power control and a slower stack temperature control. The active power control was developed by using a decentralized control method. The efficiency of the proposed hierarchical control scheme was demonstrated by case studies using the benchmark SOFC dynamic model......Based on the benchmark solid oxide fuel cell (SOFC) dynamic model for power system studies and the analysis of the SOFC operating conditions, the nonlinear programming (NLP) optimization method was used to determine the maximum electrical efficiency of the grid-connected SOFC subject...

  11. Fluorescent Nanodiamond: A Versatile Tool for Long-Term Cell Tracking, Super-Resolution Imaging, and Nanoscale Temperature Sensing.

    Hsiao, Wesley Wei-Wen; Hui, Yuen Yung; Tsai, Pei-Chang; Chang, Huan-Cheng

    2016-03-15

    Fluorescent nanodiamond (FND) has recently played a central role in fueling new discoveries in interdisciplinary fields spanning biology, chemistry, physics, and materials sciences. The nanoparticle is unique in that it contains a high density ensemble of negatively charged nitrogen-vacancy (NV(-)) centers as built-in fluorophores. The center possesses a number of outstanding optical and magnetic properties. First, NV(-) has an absorption maximum at ∼550 nm, and when exposed to green-orange light, it emits bright fluorescence at ∼700 nm with a lifetime of longer than 10 ns. These spectroscopic properties are little affected by surface modification but are distinctly different from those of cell autofluorescence and thus enable background-free imaging of FNDs in tissue sections. Such characteristics together with its excellent biocompatibility render FND ideal for long-term cell tracking applications, particularly in stem cell research. Next, as an artificial atom in the solid state, the NV(-) center is perfectly photostable, without photobleaching and blinking. Therefore, the NV-containing FND is suitable as a contrast agent for super-resolution imaging by stimulated emission depletion (STED). An improvement of the spatial resolution by 20-fold is readily achievable by using a high-power STED laser to deplete the NV(-) fluorescence. Such improvement is crucial in revealing the detailed structures of biological complexes and assemblies, including cellular organelles and subcellular compartments. Further enhancement of the resolution for live cell imaging is possible by manipulating the charge states of the NV centers. As the "brightest" member of the nanocarbon family, FND holds great promise and potential for bioimaging with unprecedented resolution and precision. Lastly, the NV(-) center in diamond is an atom-like quantum system with a total electron spin of 1. The ground states of the spins show a crystal field splitting of 2.87 GHz, separating the ms = 0 and

  12. Umbilical cord mesenchymal stem cells labeled with multimodal iron oxide nanoparticles with fluorescent and magnetic properties: application for in vivo cell tracking

    Sibov, Tatiana T; Pavon, Lorena F; Miyaki, Liza A; Mamani, Javier B; Nucci, Leopoldo P; Alvarim, Larissa T; Silveira, Paulo H; Marti, Luciana C; Gamarra, LF

    2014-01-01

    Here we describe multimodal iron oxide nanoparticles conjugated to Rhodamine-B (MION-Rh), their stability in culture medium, and subsequent validation of an in vitro protocol to label mesenchymal stem cells from umbilical cord blood (UC-MSC) with MION-Rh. These cells showed robust labeling in vitro without impairment of their functional properties, the viability of which were evaluated by proliferation kinetic and ultrastructural analyzes. Thus, labeled cells were infused into striatum of adult male rats of animal model that mimic late onset of Parkinson’s disease and, after 15 days, it was observed that cells migrated along the medial forebrain bundle to the substantia nigra as hypointense spots in T2 magnetic resonance imaging. These data were supported by short-term magnetic resonance imaging. Studies were performed in vivo, which showed that about 5 × 105 cells could be efficiently detected in the short term following infusion. Our results indicate that these labeled cells can be efficiently tracked in a neurodegenerative disease model. PMID:24531365

  13. Umbilical cord mesenchymal stem cells labeled with multimodal iron oxide nanoparticles with fluorescent and magnetic properties: application for in vivo cell tracking.

    Sibov, Tatiana T; Pavon, Lorena F; Miyaki, Liza A; Mamani, Javier B; Nucci, Leopoldo P; Alvarim, Larissa T; Silveira, Paulo H; Marti, Luciana C; Gamarra, Lf

    2014-01-01

    Here we describe multimodal iron oxide nanoparticles conjugated to Rhodamine-B (MION-Rh), their stability in culture medium, and subsequent validation of an in vitro protocol to label mesenchymal stem cells from umbilical cord blood (UC-MSC) with MION-Rh. These cells showed robust labeling in vitro without impairment of their functional properties, the viability of which were evaluated by proliferation kinetic and ultrastructural analyzes. Thus, labeled cells were infused into striatum of adult male rats of animal model that mimic late onset of Parkinson's disease and, after 15 days, it was observed that cells migrated along the medial forebrain bundle to the substantia nigra as hypointense spots in T2 magnetic resonance imaging. These data were supported by short-term magnetic resonance imaging. Studies were performed in vivo, which showed that about 5 × 10(5) cells could be efficiently detected in the short term following infusion. Our results indicate that these labeled cells can be efficiently tracked in a neurodegenerative disease model.

  14. Noninvasive in vivo tracking of mesenchymal stem cells and evaluation of cell therapeutic effects in a murine model using a clinical 3.0 T MRI.

    Drey, Florian; Choi, Yeong-Hoon; Neef, Klaus; Ewert, Birgit; Tenbrock, Arne; Treskes, Philipp; Bovenschulte, Henning; Liakopoulos, Oliver J; Brenkmann, Meike; Stamm, Christof; Wittwer, Thorsten; Wahlers, Thorsten

    2013-01-01

    Cardiac cell therapy with mesenchymal stem cells (MSCs) represents a promising treatment approach for end-stage heart failure. However, little is known about the underlying mechanisms and the fate of the transplanted cells. The objective of the presented work is to determine the feasibility of magnetic resonance imaging (MRI) and in vivo monitoring after transplantation into infarcted mouse hearts using a clinical 3.0 T MRI device. The labeling procedure of bone marrow-derived MSCs with micron-sized paramagnetic iron oxide particles (MPIOs) did not affect the viability of the cells and their cell type-defining properties when compared to unlabeled cells. Using a clinical 3.0 T MRI scanner equipped with a dedicated small animal solenoid coil, 10(5) labeled MSCs could be detected and localized in the mouse hearts for up to 4 weeks after intramyocardial transplantation. Weekly ECG-gated scans using T1-weighted sequences were performed, and left ventricular function was assessed. Histological analysis of hearts confirmed the survival of labeled MSCs in the target area up to 4 weeks after transplantation. In conclusion, in vivo tracking of labeled MSCs using a clinical 3.0 T MRI scanner is feasible. In combination with assessment of heart function, this technology allows the monitoring of the therapeutic efficacy of regenerative therapies in a small animal model.

  15. Molecular Tracking of Proteolysis During Breast Cancer Cell Extravasation: Blockage by Therapeutic Inhibitors

    Khokha, Rama

    2004-01-01

    ... (metastatic MDA- MB231 and non-metastatic MCF-7) transendothelial migration (TEM). Modulation of individual molecules demonstrates the functional cooperation of furin, cell surface adhesion molecules (alpha(sub v)Beta(sub3), CD44...

  16. MR tracking of stem cells labeled with superparamagnetic nanoparticles in ischemic brain

    Jendelová, Pavla; Růžičková, Kateřina; Urdzíková, Lucia; Kroupová, Jana; Herynek, V.; Dvořák, Petr; Hájek, M.; Syková, Eva

    č. 2 (2003), s. 35 ISSN 0894-1491. [European Meeting on Glia l Cell Function in Health and Disease /6./. Berlín, 03.09.2003-06.09.2003] R&D Projects: GA MŠk LN00A065; GA ČR GA304/03/1189 Institutional research plan: CEZ:AV0Z5039906; CEZ:MSM 111300004 Keywords : Stem cells * Nanoparticles Subject RIV: FH - Neurology Impact factor: 4.677, year: 2003

  17. Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina

    Hui Shi

    2018-04-01

    Full Text Available AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs in a mouse model of laser-induced retinal injury. METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6-carboxyfluorescein diacetate succinimidyl ester (CFSE, 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein (DiI-AcLDL, and green fluorescent protein (GFP. The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo. RESULTS: EPCs labeled with CFSE and DiI-AcLDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and DiI-AcLDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina. CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and DiI-AcLDL are suitable for short-term EPC-labeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.

  18. Chimeric Mouse model to track the migration of bone marrow derived cells in glioblastoma following anti-angiogenic treatments.

    Achyut, B R; Shankar, Adarsh; Iskander, A S M; Ara, Roxan; Knight, Robert A; Scicli, Alfonso G; Arbab, Ali S

    2016-01-01

    Bone marrow derived cells (BMDCs) have been shown to contribute in the tumor development. In vivo animal models to investigate the role of BMDCs in tumor development are poorly explored. We established a novel chimeric mouse model using as low as 5 × 10(6) GFP+ BM cells in athymic nude mice, which resulted in >70% engraftment within 14 d. In addition, chimera was established in NOD-SCID mice, which displayed >70% with in 28 d. Since anti-angiogenic therapies (AAT) were used as an adjuvant against VEGF-VEGFR pathway to normalize blood vessels in glioblastoma (GBM), which resulted into marked hypoxia and recruited BMDCs to the tumor microenvironment (TME). We exploited chimeric mice in athymic nude background to develop orthotopic U251 tumor and tested receptor tyrosine kinase inhibitors and CXCR4 antagonist against GBM. We were able to track GFP+ BMDCs in the tumor brain using highly sensitive multispectral optical imaging instrument. Increased tumor growth associated with the infiltration of GFP+ BMDCs acquiring suppressive myeloid and endothelial phenotypes was seen in TME following treatments. Immunofluorescence study showed GFP+ cells accumulated at the site of VEGF, SDF1 and PDGF expression, and at the periphery of the tumors following treatments. In conclusion, we developed a preclinical chimeric model of GBM and phenotypes of tumor infiltrated BMDCs were investigated in context of AATs. Chimeric mouse model could be used to study detailed cellular and molecular mechanisms of interaction of BMDCs and TME in cancer.

  19. "Sickle cell anemia: tracking down a mutation": an interactive learning laboratory that communicates basic principles of genetics and cellular biology.

    Jarrett, Kevin; Williams, Mary; Horn, Spencer; Radford, David; Wyss, J Michael

    2016-03-01

    "Sickle cell anemia: tracking down a mutation" is a full-day, inquiry-based, biology experience for high school students enrolled in genetics or advanced biology courses. In the experience, students use restriction endonuclease digestion, cellulose acetate gel electrophoresis, and microscopy to discover which of three putative patients have the sickle cell genotype/phenotype using DNA and blood samples from wild-type and transgenic mice that carry a sickle cell mutation. The inquiry-based, problem-solving approach facilitates the students' understanding of the basic concepts of genetics and cellular and molecular biology and provides experience with contemporary tools of biotechnology. It also leads to students' appreciation of the causes and consequences of this genetic disease, which is relatively common in individuals of African descent, and increases their understanding of the first principles of genetics. This protocol provides optimal learning when led by well-trained facilitators (including the classroom teacher) and carried out in small groups (6:1 student-to-teacher ratio). This high-quality experience can be offered to a large number of students at a relatively low cost, and it is especially effective in collaboration with a local science museum and/or university. Over the past 15 yr, >12,000 students have completed this inquiry-based learning experience and demonstrated a consistent, substantial increase in their understanding of the disease and genetics in general. Copyright © 2016 The American Physiological Society.

  20. A study of V79 cell survival after for proton and carbon ion beams as represented by the parameters of Katz' track structure model

    Grzanka, Leszek; Waligórski, M. P. R.; Bassler, Niels

    different sets of data obtained for the same cell line and different ions, measured at different laboratories, we have fitted model parameters to a set of carbon-irradiated V79 cells, published by Furusawa et al. (2), and to a set of proton-irradiated V79 cells, published by Wouters et al. (3), separately....... We found that values of model parameters best fitted to the carbon data of Furusawa et al. yielded predictions of V79 survival after proton irradiation which did not match the V79 proton data of Wouters et al. Fitting parameters to both sets combined did not improve the accuracy of model predictions...... carbon irradiation. 1. Katz, R., Track structure in radiobiology and in radiation detection. Nuclear Track Detection 2: 1-28 (1978). 2. Furusawa Y. et al. Inactivation of aerobic and hypoxic cells from three different cell lines by accelerated 3He-, 12C- and 20Ne beams. Radiat Res. 2012 Jan; 177...

  1. Thyroid cell irradiation by radioiodines: a new Monte Carlo electron track-structure code

    Champion, Christophe; Elbast, Mouhamad; Colas-Linhart, Nicole; Ting-Di Wu

    2007-01-01

    The most significant impact of the Chernobyl accident is the increased incidence of thyroid cancer among children who were exposed to short-lived radioiodines and 131-iodine. In order to accurately estimate the radiation dose provided by these radioiodines, it is necessary to know where iodine is incorporated. To do that, the distribution at the cellular level of newly organified iodine in the immature rat thyroid was performed using secondary ion mass microscopy (NanoSIMS 50 ). Actual dosimetric models take only into account the averaged energy and range of beta particles of the radio-elements and may, therefore, imperfectly describe the real distribution of dose deposit at the microscopic level around the point sources. Our approach is radically different since based on a track-structure Monte Carlo code allowing following-up of electrons down to low energies (∼= 10 eV) what permits a nanometric description of the irradiation physics. The numerical simulations were then performed by modelling the complete disintegrations of the short-lived iodine isotopes as well as of 131 I in new born rat thyroids in order to take into account accurate histological and biological data for the thyroid gland. (author)

  2. Using Sunlight and Cell Networks to Bring Fleeting Tracking to Small Scale Fisheries

    Garren, M.; Selbie, H.; Suchomel, D.; McDonald, W.; Solomon, D.

    2016-12-01

    Traditionally, the efforts of small scale fisheries have not been easily incorporated into the global picture of fishing effort and activity. That means that the activities of the vast majority ( 90%) of fishing vessels in the world have remained unquantified and largely opaque. With newly developed technology that harnesses solar power and cost-effective cellular networks to transmit data, it is becoming possible to provide vessel tracking systems on a large scale for vessels of all sizes. Furthermore, capitalizing on the relatively inexpensive cellular networks to transfer the data enables data of much higher granularity to be captured. By recording a vessel's position every few seconds, instead of minutes to hours as is typical of most satellite-based systems, we are able to resolve a diverse array of behaviors happening at sea including when and where fishing occurred and what type of fishing gear was used. This high granularity data is both incredibly useful and also a challenge to manage and mine. New approaches for handling and processing this continuous data stream of vessel positions are being developed to extract the most informative and actionable pieces of information for a variety of audiences including governing agencies, industry supply chains seeking transparency, non-profit organizations supporting conservation efforts, academic researchers and the fishers and boat owners.

  3. Full-Volume, Three-Dimensional, Transient Measurements of Bubbly Flows Using Particle Tracking Velocimetry and Shadow Image Velocimetry Coupled with Pattern Recognition Techniques

    Yassin Hassan

    2001-01-01

    Develop a state-of-the-art non-intrusive diagnostic tool to perform simultaneous measurements of both the temporal and three-dimensional spatial velocity of the two phases of a bubbly flow. These measurements are required to provide a foundation for studying the constitutive closure relations needed in computational fluid dynamics and best-estimate thermal hydraulic codes employed in nuclear reactor safety analysis and severe accident simulation. Such kinds of full-field measurements are not achievable through the commonly used point-measurement techniques, such as hot wire, conductance probe, laser Doppler anemometry, etc. The results can also be used in several other applications, such as the dynamic transport of pollutants in water or studies of the dispersion of hazardous waste

  4. Germline Transgenic Methods for Tracking Cells and Testing Gene Function during Regeneration in the Axolotl

    Khattak, Shahryar; Schuez, Maritta; Richter, Tobias; Knapp, Dunja; Haigo, Saori L.; Sandoval-Guzmán, Tatiana; Hradlikova, Kristyna; Duemmler, Annett; Kerney, Ryan; Tanaka, Elly M.

    2013-01-01

    The salamander is the only tetrapod that regenerates complex body structures throughout life. Deciphering the underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but the model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in the laboratory-bred salamander Ambystoma mexicanum (axolotl) that open up the cellular and molecular genetic dissection of regeneration. We demonstrate tissue-dependent control of gene expression in nerve, Schwann cells, oligodendrocytes, muscle, epidermis, and cartilage. Furthermore, we demonstrate the use of tamoxifen-induced Cre/loxP-mediated recombination to indelibly mark different cell types. Finally, we inducibly overexpress the cell-cycle inhibitor p16INK4a, which negatively regulates spinal cord regeneration. These tissue-specific germline axolotl lines and tightly inducible Cre drivers and LoxP reporter lines render this classical regeneration model molecularly accessible. PMID:24052945

  5. Cu2ZnSnS4 solar cells: Physics and technology by alternative tracks

    Crovetto, Andrea

    things: i) alternative solar absorbers (notably, Cu2SnS3) that are chemically related to CZTS and that have similar selling points; ii) other materials included in the device stack of CZTS solar cells. Here I list what I believe the main highlights of my work are. First, we achieve the highest reported...... power conversion eciency (5.2%) for a CZTS solar cell using pulsed laser deposition as a fabrication method for CZTS precursors. This is thanks to to joint work of PhD student Andrea Cazzaniga, PhD student Chang Yan (University of New South Wales, Australia) and myself. Perhaps more importantly, we...... finally understand, albeit very roughly, the "rules of the game" for successful pulsed laser deposition of high-quality chalcogenide precursors for solar cells. This kind of understanding is not evident in the existing literature and is mostly the result of the work of PhD student Andrea Cazzaniga. Second...

  6. Particle image velocimetry and infrared thermography in a levitated droplet with nanosilica suspensions

    Saha, Abhishek; Kumar, Ranganathan [University of Central Florida, Department of Mechanical Materials and Aerospace Engineering, Orlando, FL (United States); Basu, Saptarshi [Indian Institute of Science, Department of Mechanical Engineering, Bangalore (India)

    2012-03-15

    Preferential accumulation and agglomeration kinetics of nanoparticles suspended in an acoustically levitated water droplet under radiative heating has been studied. Particle image velocimetry performed to map the internal flow field shows a single cell recirculation with increasing strength for decreasing viscosities. Infrared thermography and high speed imaging show details of the heating process for various concentrations of nanosilica droplets. Initial stage of heating is marked by fast vaporization of liquid and sharp temperature rise. Following this stage, aggregation of nanoparticles is seen resulting in various structure formations. At low concentrations, a bowl structure of the droplet is dominant, maintained at a constant temperature. At high concentrations, viscosity of the solution increases, leading to rotation about the levitator axis due to the dominance of centrifugal motion. Such complex fluid motion inside the droplet due to acoustic streaming eventually results in the formation of a ring structure. This horizontal ring eventually reorients itself due to an imbalance of acoustic forces on the ring, exposing larger area for laser absorption and subsequent sharp temperature rise. (orig.)

  7. A tomographic particle image velocimetry investigation of the flow development over dual step cylinders

    Morton, C., E-mail: chris.morton@ucalgary.ca [Department of Mechanical and Manufacturing Engineering, University of Calgary, 2500 University Dr NW, Calgary, Alberta T2N 1N4 (Canada); Yarusevych, S. [Department of Mechanical and Mechatronics Engineering, University of Waterloo, 200 University Ave W, Waterloo, Ontario N2L 3G1 (Canada); Scarano, F. [Department of Aerospace Engineering, Delft University of Technology, 2628 Delft (Netherlands)

    2016-02-15

    This experimental study focuses on the near wake development of a dual step cylinder geometry consisting of a long base cylinder of diameter d to which a larger diameter (D) cylinder of length L is attached coaxially at mid-span. The experiments cover a range of Reynolds numbers, 2000 ≤ Re{sub D} ≤ 5000, diameter ratios, 1.33 ≤ D/d ≤ 2.0 and large cylinder aspect ratios, 0.5 ≤ L/D ≤ 5 using Tomographic particle image velocimetry. Distinct changes in wake topology are observed varying the above parameters. Supporting previous experimental studies on the same geometry involving flow visualization and planar measurements, four distinct flow regimes are identified to which a distinct three-dimensional wake topology can be associated. The vortex-dominated wake dynamical behaviour is investigated with Proper Orthogonal Decomposition (POD) and conditional averaging of three-dimensional velocity fields is used to exemplify the different shedding regimes. The conditionally averaged flow fields are shown to quantitatively resolve flow features equivalent to those obtained from a reduced order model consisting of the first ten to twenty POD modes, identifying the dominant vortex shedding cells and their interactions.

  8. Investigation of the hydrodynamic behavior of diatom aggregates using particle image velocimetry.

    Xiao, Feng; Li, Xiaoyan; Lam, Kitming; Wang, Dongsheng

    2012-01-01

    The hydrodynamic behavior of diatom aggregates has a significant influence on the interactions and flocculation kinetics of algae. However, characterization of the hydrodynamics of diatoms and diatom aggregates in water is rather difficult. In this laboratory study, an advanced visualization technique in particle image velocimetry (PIV) was employed to investigate the hydrodynamic properties of settling diatom aggregates. The experiments were conducted in a settling column filled with a suspension of fluorescent polymeric beads as seed tracers. A laser light sheet was generated by the PIV setup to illuminate a thin vertical planar region in the settling column, while the motions of particles were recorded by a high speed charge-coupled device (CCD) camera. This technique was able to capture the trajectories of the tracers when a diatom aggregate settled through the tracer suspension. The PIV results indicated directly the curvilinear feature of the streamlines around diatom aggregates. The rectilinear collision model largely overestimated the collision areas of the settling particles. Algae aggregates appeared to be highly porous and fractal, which allowed streamlines to penetrate into the aggregate interior. The diatom aggregates have a fluid collection efficiency of 10%-40%. The permeable feature of aggregates can significantly enhance the collisions and flocculation between the aggregates and other small particles including algal cells in water.

  9. Cell tracking in cardiac repair: What to image and how to image

    A. Ruggiero (Alessandro); D.L.J. Thorek (Daniel L.J.); J. Guenoun (Jamal); G.P. Krestin (Gabriel); M.R. Bernsen (Monique)

    2012-01-01

    textabstractStem cell therapies hold the great promise and interest for cardiac regeneration among scientists, clinicians and patients. However, advancement and distillation of a standard treatment regimen are not yet finalised. Into this breach step recent developments in the imaging biosciences.

  10. Labeling human embryonic stem-cell-derived cardiomyocytes for tracking with MR imaging

    Castaneda, Rosalinda T.; Daldrup-Link, Heike [Lucile Packard Children' s Hospital, Stanford School of Medicine, Pediatric Radiology, Stanford, CA (United States); Boddington, Sophie; Wendland, Mike; Mandrussow, Lydia [University of California, Department of Radiology and Biomedical Imaging, UCSF Medical Center, San Francisco, CA (United States); Henning, Tobias D. [University Hospital of Cologne, Department of Radiology and Neuroradiology, Cologne (Germany); Liu, Siyuan [National Institutes of Health, Language Section, Voice, Speech and Language Branch, National Institute on Deafness and Other Communication Disorders, Bethesda, MD (United States)

    2011-11-15

    Human embryonic stem cells (hESC) can generate cardiomyocytes (CM), which offer promising treatments for cardiomyopathies in children. However, challenges for clinical translation result from loss of transplanted cell from target sites and high cell death. An imaging technique that noninvasively and repetitively monitors transplanted hESC-CM could guide improvements in transplantation techniques and advance therapies. To develop a clinically applicable labeling technique for hESC-CM with FDA-approved superparamagnetic iron oxide nanoparticles (SPIO) by examining labeling before and after CM differentiation. Triplicates of hESC were labeled by simple incubation with 50 {mu}g/ml of ferumoxides before or after differentiation into CM, then imaged on a 7T MR scanner using a T2-weighted multi-echo spin-echo sequence. Viability, iron uptake and T2-relaxation times were compared between groups using t-tests. hESC-CM labeled before differentiation demonstrated significant MR effects, iron uptake and preserved function. hESC-CM labeled after differentiation showed no significant iron uptake or change in MR signal (P < 0.05). Morphology, differentiation and viability were consistent between experimental groups. hESC-CM should be labeled prior to CM differentiation to achieve a significant MR signal. This technique permits monitoring delivery and engraftment of hESC-CM for potential advancements of stem cell-based therapies in the reconstitution of damaged myocardium. (orig.)

  11. Genome-wide tracking of unmethylated DNA Alu repeats in normal and cancer cells

    Rodriguez, Jairo; Vives, Laura; Jordà, Mireia

    2008-01-01

    Methylation of the cytosine is the most frequent epigenetic modification of DNA in mammalian cells. In humans, most of the methylated cytosines are found in CpG-rich sequences within tandem and interspersed repeats that make up to 45% of the human genome, being Alu repeats the most common family....

  12. Labeling human embryonic stem-cell-derived cardiomyocytes for tracking with MR imaging

    Castaneda, Rosalinda T.; Daldrup-Link, Heike; Boddington, Sophie; Wendland, Mike; Mandrussow, Lydia; Henning, Tobias D.; Liu, Siyuan

    2011-01-01

    Human embryonic stem cells (hESC) can generate cardiomyocytes (CM), which offer promising treatments for cardiomyopathies in children. However, challenges for clinical translation result from loss of transplanted cell from target sites and high cell death. An imaging technique that noninvasively and repetitively monitors transplanted hESC-CM could guide improvements in transplantation techniques and advance therapies. To develop a clinically applicable labeling technique for hESC-CM with FDA-approved superparamagnetic iron oxide nanoparticles (SPIO) by examining labeling before and after CM differentiation. Triplicates of hESC were labeled by simple incubation with 50 μg/ml of ferumoxides before or after differentiation into CM, then imaged on a 7T MR scanner using a T2-weighted multi-echo spin-echo sequence. Viability, iron uptake and T2-relaxation times were compared between groups using t-tests. hESC-CM labeled before differentiation demonstrated significant MR effects, iron uptake and preserved function. hESC-CM labeled after differentiation showed no significant iron uptake or change in MR signal (P < 0.05). Morphology, differentiation and viability were consistent between experimental groups. hESC-CM should be labeled prior to CM differentiation to achieve a significant MR signal. This technique permits monitoring delivery and engraftment of hESC-CM for potential advancements of stem cell-based therapies in the reconstitution of damaged myocardium. (orig.)

  13. The Impact of the Geometrical Structure of the DNA on Parameters of the Track-Event Theory for Radiation Induced Cell Kill.

    Uwe Schneider

    Full Text Available When fractionation schemes for hypofractionation and stereotactic body radiotherapy are considered, a reliable cell survival model at high dose is needed for calculating doses of similar biological effectiveness. An alternative to the LQ-model is the track-event theory which is based on the probabilities for one- and two two-track events. A one-track-event (OTE is always represented by at least two simultaneous double strand breaks. A two-track-event (TTE results in one double strand break. Therefore at least two two-track-events on the same or different chromosomes are necessary to produce an event which leads to cell sterilization. It is obvious that the probabilities of OTEs and TTEs must somehow depend on the geometrical structure of the chromatin. In terms of the track-event theory the ratio ε of the probabilities of OTEs and TTEs includes the geometrical dependence and is obtained in this work by simple Monte Carlo simulations.For this work it was assumed that the anchors of loop forming chromatin are most sensitive to radiation induced cell deaths. Therefore two adjacent tetranucleosomes representing the loop anchors were digitized. The probability ratio ε of OTEs and TTEs was factorized into a radiation quality dependent part and a geometrical part: ε = εion ∙ εgeo. εgeo was obtained for two situations, by applying Monte Carlo simulation for DNA on the tetranucleosomes itself and for linker DNA. Low energy electrons were represented by randomly distributed ionizations and high energy electrons by ionizations which were simulated on rays. εion was determined for electrons by using results from nanodosimetric measurements. The calculated ε was compared to the ε obtained from fits of the track event model to 42 sets of experimental human cell survival data.When the two tetranucleosomes are in direct contact and the hits are randomly distributed εgeo and ε are 0.12 and 0.85, respectively. When the hits are simulated on rays

  14. Cryogenic flow rate measurement with a laser Doppler velocimetry standard

    Maury, R.; Strzelecki, A.; Auclercq, C.; Lehot, Y.; Loubat, S.; Chevalier, J.; Ben Rayana, F.

    2018-03-01

    A very promising alternative to the state-of-the-art static volume measurements for liquefied natural gas (LNG) custody transfer processes is the dynamic principle of flow metering. As the Designated Institute (DI) of the LNE (‘Laboratoire National de métrologie et d’Essais’, being the French National Metrology Institute) for high-pressure gas flow metering, Cesame-Exadebit is involved in various research and development programs. Within the framework of the first (2010-2013) and second (2014-2017) EURAMET Joint Research Project (JRP), named ‘Metrological support for LNG custody transfer and transport fuel applications’, Cesame-Exadebit explored a novel cryogenic flow metering technology using laser Doppler velocimetry (LDV) as an alternative to ultrasonic and Coriolis flow metering. Cesame-Exadebit is trying to develop this technique as a primary standard for cryogenic flow meters. Currently, cryogenic flow meters are calibrated at ambient temperatures with water. Results are then extrapolated to be in the Reynolds number range of real applications. The LDV standard offers a unique capability to perform online calibration of cryogenic flow meters in real conditions (temperature, pressure, piping and real flow disturbances). The primary reference has been tested on an industrial process in a LNG terminal during truck refuelling. The reference can calibrate Coriolis flow meters being used daily with all the real environmental constraints, and its utilisation is transparent for LNG terminal operators. The standard is traceable to Standard International units and the combined extended uncertainties have been determined and estimated to be lower than 0.6% (an ongoing improvement to reducing the correlation function uncertainty, which has a major impact in the uncertainty estimation).

  15. Digital particle image thermometry/velocimetry: a review

    Dabiri, Dana [University of Washington, Department of Aeronautics and Astronautics, Seattle, WA (United States)

    2009-02-15

    Digital particle image thermometry/velocimetry (DPIT/V) is a relatively new methodology that allows for measurements of simultaneous temperature and velocity within a two-dimensional domain, using thermochromic liquid crystal tracer particles as the temperature and velocity sensors. Extensive research has been carried out over recent years that have allowed the methodology and its implementation to grow and evolve. While there have been several reviews on the topic of liquid crystal thermometry (Moffat in Exp Therm Fluid Sci 3:14-32, 1990; Baughn in Int J Heat Fluid Flow 16:365-375, 1995; Roberts and East in J Spacecr Rockets 33:761-768, 1996; Wozniak et al. in Appl Sci Res 56:145-156, 1996; Behle et al. in Appl Sci Res 56:113-143, 1996; Stasiek in Heat Mass Transf 33:27-39, 1997; Stasiek and Kowalewski in Opto Electron Rev 10:1-10, 2002; Stasiek et al. in Opt Laser Technol 38:243-256, 2006; Smith et al. in Exp Fluids 30:190-201, 2001; Kowalewski et al. in Springer handbook of experimental fluid mechanics, 1st edn. Springer, Berlin, pp 487-561, 2007), the focus of the present review is to provide a relevant discussion of liquid crystals pertinent to DPIT/V. This includes a background on liquid crystals and color theory, a discussion of experimental setup parameters, a description of the methodology's most recent advances and processing methods affecting temperature measurements, and finally an explanation of its various implementations and applications. (orig.)

  16. Peak-locking reduction for particle image velocimetry

    Michaelis, Dirk; Wieneke, Bernhard; Neal, Douglas R

    2016-01-01

    A parametric study of the factors contributing to peak-locking, a known bias error source in particle image velocimetry (PIV), is conducted using synthetic data that are processed with a state-of-the-art PIV algorithm. The investigated parameters include: particle image diameter, image interpolation techniques, the effect of asymmetric versus symmetric window deformation, number of passes and the interrogation window size. Some of these parameters are found to have a profound effect on the magnitude of the peak-locking error. The effects for specific PIV cameras are also studied experimentally using a precision turntable to generate a known rotating velocity field. Image time series recorded using this experiment show a linear range of pixel and sub-pixel shifts ranging from 0 to  ±4 pixels. Deviations in the constant vorticity field (ω z ) reveal how peak-locking can be affected systematically both by varying parameters of the detection system such as the focal distance and f -number, and also by varying the settings of the PIV analysis. A new a priori technique for reducing the bias errors associated with peak-locking in PIV is introduced using an optical diffuser to avoid undersampled particle images during the recording of the raw images. This technique is evaluated against other a priori approaches using experimental data and is shown to perform favorably. Finally, a new a posteriori anti peak-locking filter (APLF) is developed and investigated, which shows promising results for both synthetic data and real measurements for very small particle image sizes. (paper)

  17. Poly (dopamine) coated superparamagnetic iron oxide nanocluster for noninvasive labeling, tracking, and targeted delivery of adipose tissue-derived stem cells

    Liao, Naishun; Wu, Ming; Pan, Fan; Lin, Jiumao; Li, Zuanfang; Zhang, Da; Wang, Yingchao; Zheng, Youshi; Peng, Jun; Liu, Xiaolong; Liu, Jingfeng

    2016-01-01

    Tracking and monitoring of cells in vivo after transplantation can provide crucial information for stem cell therapy. Magnetic resonance imaging (MRI) combined with contrast agents is believed to be an effective and non-invasive technique for cell tracking in living bodies. However, commercial superparamagnetic iron oxide nanoparticles (SPIONs) applied to label cells suffer from shortages such as potential toxicity, low labeling efficiency, and low contrast enhancing. Herein, the adipose tissue-derived stem cells (ADSCs) were efficiently labeled with SPIONs coated with poly (dopamine) (SPIONs cluster@PDA), without affecting their viability, proliferation, apoptosis, surface marker expression, as well as their self-renew ability and multi-differentiation potential. The labeled cells transplanted into the mice through tail intravenous injection exhibited a negative enhancement of the MRI signal in the damaged liver-induced by carbon tetrachloride, and subsequently these homed ADSCs with SPIONs cluster@PDA labeling exhibited excellent repair effects to the damaged liver. Moreover, the enhanced target-homing to tissue of interest and repair effects of SPIONs cluster@PDA-labeled ADSCs could be achieved by use of external magnetic field in the excisional skin wound mice model. Therefore, we provide a facile, safe, noninvasive and sensitive method for external magnetic field targeted delivery and MRI based tracking of transplanted cells in vivo.

  18. Poly (dopamine) coated superparamagnetic iron oxide nanocluster for noninvasive labeling, tracking, and targeted delivery of adipose tissue-derived stem cells.

    Liao, Naishun; Wu, Ming; Pan, Fan; Lin, Jiumao; Li, Zuanfang; Zhang, Da; Wang, Yingchao; Zheng, Youshi; Peng, Jun; Liu, Xiaolong; Liu, Jingfeng

    2016-01-05

    Tracking and monitoring of cells in vivo after transplantation can provide crucial information for stem cell therapy. Magnetic resonance imaging (MRI) combined with contrast agents is believed to be an effective and non-invasive technique for cell tracking in living bodies. However, commercial superparamagnetic iron oxide nanoparticles (SPIONs) applied to label cells suffer from shortages such as potential toxicity, low labeling efficiency, and low contrast enhancing. Herein, the adipose tissue-derived stem cells (ADSCs) were efficiently labeled with SPIONs coated with poly (dopamine) (SPIONs cluster@PDA), without affecting their viability, proliferation, apoptosis, surface marker expression, as well as their self-renew ability and multi-differentiation potential. The labeled cells transplanted into the mice through tail intravenous injection exhibited a negative enhancement of the MRI signal in the damaged liver-induced by carbon tetrachloride, and subsequently these homed ADSCs with SPIONs cluster@PDA labeling exhibited excellent repair effects to the damaged liver. Moreover, the enhanced target-homing to tissue of interest and repair effects of SPIONs cluster@PDA-labeled ADSCs could be achieved by use of external magnetic field in the excisional skin wound mice model. Therefore, we provide a facile, safe, noninvasive and sensitive method for external magnetic field targeted delivery and MRI based tracking of transplanted cells in vivo.

  19. Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking

    Ece Akhan

    2015-01-01

    Full Text Available Mesenchymal stem cells (MSCs are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.

  20. Tracking the Oxygen Status in the Cell Nucleus with a Hoechst-Tagged Phosphorescent Ruthenium Complex.

    Hara, Daiki; Umehara, Yui; Son, Aoi; Asahi, Wataru; Misu, Sotaro; Kurihara, Ryohsuke; Kondo, Teruyuki; Tanabe, Kazuhito

    2018-05-04

    Molecular oxygen in living cells is distributed and consumed inhomogeneously, depending on the activity of each organelle. Therefore, tractable methods that can be used to monitor the oxygen status in each organelle are needed to understand cellular function. Here we report the design of a new oxygen-sensing probe for use in the cell nucleus. We prepared "Ru-Hoechsts", each consisting of a phosphorescent ruthenium complex linked to a Hoechst 33258 moiety, and characterized their properties as oxygen sensors. The Hoechst unit shows strong DNA-binding properties in the nucleus, and the ruthenium complex shows oxygen-dependent phosphorescence. Thus, Ru-Hoechsts accumulated in the cell nucleus and showed oxygen-dependent signals that could be monitored. Of the Ru-Hoechsts prepared in this study, Ru-Hoechst b, in which the ruthenium complex and the Hoechst unit were linked through a hexyl chain, showed the most suitable properties for monitoring the oxygen status. Ru-Hoechsts are probes with high potential for visualizing oxygen fluctuations in the nucleus. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Tracking cell surface mobility of GPCRs using α-bungarotoxin-linked fluorophores.

    Hannan, Saad; Wilkins, Megan E; Thomas, Philip; Smart, Trevor G

    2013-01-01

    GABA(B) receptors are G-protein-coupled receptors (GPCRs) that are activated by GABA, the principal inhibitory neurotransmitter in the central nervous system. Cell surface mobility of GABA(B) receptors is a key determinant of the efficacy of slow and prolonged synaptic inhibition initiated by GABA. Therefore, experimentally monitoring receptor mobility and how this can be regulated is of primary importance for understanding the roles of GABA(B) receptors in the brain, and how they may be therapeutically exploited. Unusually for a GPCR, heterodimerization between the R1 and R2 subunits is required for the cell surface expression and signaling by prototypical GABA(B) receptors. Here, we describe a minimal epitope-tagging method, based on the incorporation of an α-bungarotoxin binding site (BBS) into the GABA(B) receptor, to study receptor internalization in live cells using a range of imaging approaches. We demonstrate how this technique can be adapted by modifying the BBS to monitor the simultaneous movement of both R1 and R2 subunits, revealing that GABA(B) receptors are internalized as heteromers. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. SU-D-210-04: Using Radiotherapy Biomaterials to Brand and Track Deadly Cancer Cells

    Altundal, Y; Sajo, E [Univ Massachusetts Lowell, Lowell, MA (United States); Ngwa, W [Univ Massachusetts Lowell, Lowell, MA (United States); Brigham and Women’s Hospital, Dana Farber Cancer Institute, Harvard Medical, Boston, MA (United States)

    2015-06-15

    Purpose: Metastasis accounts for over 90% of all cancer associated suffering and death and arguably presents the most formidable challenges in cancer management. The detection of metastatic or rare circulating tumor cells (CTCs) in blood or lymph nodes remains a formidable technological challenge. In this study, we investigated the time needed to label each cancer cell in-situ (right at the source tumor) with sufficient number of GNPs that will allow enhanced non-invasive detection via photoacoustic imaging in the lymph nodes. Such in-situ labeling can be achieved via sustained release of the GNPs from Radiotherapy (RT) biomaterials (e.g. fiducials, spacers) coated/loaded with the GNP. Methods: The minimum concentration (1000 GNPs/cell for 50nm GNPs) to detect GNPs with photoacoustic imaging method was experimentally measured by Mallidi et al. and fixed at the tumor sub-volume periphery. In this work, the GNPs were assumed to diffuse from a point source, placed in the middle of a 2–3cm tumor, with an initial concentration of 7–30 mg/g. The time required to label the cells with GNPs was calculated by solving the three dimensional diffusion-reaction equation analytically. The diffusion coefficient of 10nm GNPs was experimentally determined previously. Stokes-Einstein equation was used to calculate the diffusion coefficients for other sizes (2–50nm) of GNPs. The cellular uptake rate constants for several sizes of GNPs were experimentally measured by Jin et al. Results: The time required to label the cells was found 0.635–15.91 days for 2–50nm GNPs with an initial concentration of 7 mg/g GNPs in a 2 cm tumor; 1.379–34.633 days for 2–50nm GNPs with an initial concentration of 30 mg/g GNPs in a 3cm tumor. Conclusion: Our results highlight new potential for labeling CTCs with GNPs released from smart RT biomaterials (i.e. fiducials or spacers loaded with the GNP) towards enhanced non-invasive imaging/detection via photoacoustic imaging.

  3. SU-D-210-04: Using Radiotherapy Biomaterials to Brand and Track Deadly Cancer Cells

    Altundal, Y; Sajo, E; Ngwa, W

    2015-01-01

    Purpose: Metastasis accounts for over 90% of all cancer associated suffering and death and arguably presents the most formidable challenges in cancer management. The detection of metastatic or rare circulating tumor cells (CTCs) in blood or lymph nodes remains a formidable technological challenge. In this study, we investigated the time needed to label each cancer cell in-situ (right at the source tumor) with sufficient number of GNPs that will allow enhanced non-invasive detection via photoacoustic imaging in the lymph nodes. Such in-situ labeling can be achieved via sustained release of the GNPs from Radiotherapy (RT) biomaterials (e.g. fiducials, spacers) coated/loaded with the GNP. Methods: The minimum concentration (1000 GNPs/cell for 50nm GNPs) to detect GNPs with photoacoustic imaging method was experimentally measured by Mallidi et al. and fixed at the tumor sub-volume periphery. In this work, the GNPs were assumed to diffuse from a point source, placed in the middle of a 2–3cm tumor, with an initial concentration of 7–30 mg/g. The time required to label the cells with GNPs was calculated by solving the three dimensional diffusion-reaction equation analytically. The diffusion coefficient of 10nm GNPs was experimentally determined previously. Stokes-Einstein equation was used to calculate the diffusion coefficients for other sizes (2–50nm) of GNPs. The cellular uptake rate constants for several sizes of GNPs were experimentally measured by Jin et al. Results: The time required to label the cells was found 0.635–15.91 days for 2–50nm GNPs with an initial concentration of 7 mg/g GNPs in a 2 cm tumor; 1.379–34.633 days for 2–50nm GNPs with an initial concentration of 30 mg/g GNPs in a 3cm tumor. Conclusion: Our results highlight new potential for labeling CTCs with GNPs released from smart RT biomaterials (i.e. fiducials or spacers loaded with the GNP) towards enhanced non-invasive imaging/detection via photoacoustic imaging

  4. Umbilical cord mesenchymal stem cells labeled with multimodal iron oxide nanoparticles with fluorescent and magnetic properties: application for in vivo cell tracking

    Sibov TT

    2014-01-01

    Full Text Available Tatiana T Sibov,1,2 Lorena F Pavon,1 Liza A Miyaki,1 Javier B Mamani,1 Leopoldo P Nucci,1,2 Larissa T Alvarim,1,3 Paulo H Silveira,1 Luciana C Marti,1 LF Gamarra1–31Hospital Israelita Albert Einstein, São Paulo, Brazil; 2Departamento de Neurologia e Neurociências, Universidade Federal de São Paulo, São Paulo, Brazil; 3Faculdade de Ciências Médicas da Santa Casa de São Paulo, São Paulo, BrazilAbstract: Here we describe multimodal iron oxide nanoparticles conjugated to Rhodamine-B (MION-Rh, their stability in culture medium, and subsequent validation of an in vitro protocol to label mesenchymal stem cells from umbilical cord blood (UC-MSC with MION-Rh. These cells showed robust labeling in vitro without impairment of their functional properties, the viability of which were evaluated by proliferation kinetic and ultrastructural analyzes. Thus, labeled cells were infused into striatum of adult male rats of animal model that mimic late onset of Parkinson's disease and, after 15 days, it was observed that cells migrated along the medial forebrain bundle to the substantia nigra as hypointense spots in T2 magnetic resonance imaging. These data were supported by short-term magnetic resonance imaging. Studies were performed in vivo, which showed that about 5 × 105 cells could be efficiently detected in the short term following infusion. Our results indicate that these labeled cells can be efficiently tracked in a neurodegenerative disease model.Keywords: mesenchymal stem cells, multimodal iron oxide nanoparticles, Rhodamine, magnetic resonance imaging, Parkinson's disease

  5. Cell tracking and therapy evaluation of bone marrow monocytes and stromal cells using SPECT and CMR in a canine model of myocardial infarction

    Merrifield Peter

    2009-04-01

    Full Text Available Abstract Background The clinical application of stem cell therapy for myocardial infarction will require the development of methods to monitor treatment and pre-clinical assessment in a large animal model, to determine its effectiveness and the optimum cell population, route of delivery, timing, and flow milieu. Objectives To establish a model for a in vivo tracking to monitor cell engraftment after autologous transplantation and b concurrent measurement of infarct evolution and remodeling. Methods We evaluated 22 dogs (8 sham controls, 7 treated with autologous bone marrow monocytes, and 7 with stromal cells using both imaging of 111Indium-tropolone labeled cells and late gadolinium enhancement CMR for up to12 weeks after a 3 hour coronary occlusion. Hearts were also examined using immunohistochemistry for capillary density and presence of PKH26 labeled cells. Results In vivo Indium imaging demonstrated an effective biological clearance half-life from the injection site of ~5 days. CMR demonstrated a pattern of progressive infarct shrinkage over 12 weeks, ranging from 67–88% of baseline values with monocytes producing a significant treatment effect. Relative infarct shrinkage was similar through to 6 weeks in all groups, following which the treatment effect was manifest. There was a trend towards an increase in capillary density with cell treatment. Conclusion This multi-modality approach will allow determination of the success and persistence of engraftment, and a correlation of this with infarct size shrinkage, regional function, and left ventricular remodeling. There were overall no major treatment effects with this particular model of transplantation immediately post-infarct.

  6. Simultaneous Multi-Species Tracking in Live Cells with Quantum Dot Conjugates

    Clausen, M. P.; Christensen, Eva Arnspang; Ballou, B.

    2014-01-01

    Quantum dots are available in a range of spectrally separated emission colors and with a range of water-stabilizing surface coatings that offers great flexibility for enabling bio-specificity. In this study, we have taken advantage of this flexibility to demonstrate that it is possible to perform...... a simultaneous investigation of the lateral dynamics in the plasma membrane of i) the transmembrane epidermal growth factor receptor, ii) the glucosylphospatidylinositol-anchored protein CD59, and iii) ganglioside G(M1)-cholera toxin subunit B clusters in a single cell. We show that a large number...

  7. FAST TRACK COMMUNICATION: Eight-logic memory cell based on multiferroic junctions

    Yang, Feng; Zhou, Y. C.; Tang, M. H.; Liu, Fen; Ma, Ying; Zheng, X. J.; Zhao, W. F.; Xu, H. Y.; Sun, Z. H.

    2009-04-01

    A model is proposed for a device combining a multiferroic tunnel junction with a magnetoelectric (ME) film in which the magnetic configuration is controlled by the electric field. Calculations embodying the Green's function approach show that the magnetic polarization can be switched on and off by an electric field in the ME film due to the effect of elastic coupling interaction. Using a model including the spin-filter effect and screening of polarization charges, we have produced eight logic states of tunnelling resistance in the tunnel junction and have obtained corresponding laws that control them. The results provide some insights into the realization of an eight-logic memory cell.

  8. Superparamagnetic iron oxide nanoparticle-labeled cells as an effective vehicle for tracking the GFP gene marker using magnetic resonance imaging

    Zhang, Z; Mascheri, N; Dharmakumar, R; Fan, Z; Paunesku, T; Woloschak, G; Li, D

    2010-01-01

    Background Detection of a gene using magnetic resonance imaging (MRI) is hindered by the magnetic resonance (MR) targeting gene technique. Therefore it may be advantageous to image gene-expressing cells labeled with superparamagnetic iron oxide (SPIO) nanoparticles by MRI. Methods The GFP-R3230Ac (GFP) cell line was incubated for 24 h using SPIO nanoparticles at a concentration of 20 μg Fe/mL. Cell samples were prepared for iron content analysis and cell function evaluation. The labeled cells were imaged using fluorescent microscopy and MRI. Results SPIO was used to label GFP cells effectively, with no effects on cell function and GFP expression. Iron-loaded GFP cells were successfully imaged with both fluorescent microscopy and T2*-weighted MRI. Prussian blue staining showed intracellular iron accumulation in the cells. All cells were labeled (100% labeling efficiency). The average iron content per cell was 4.75±0.11 pg Fe/cell (P<0.05 versus control). Discussion This study demonstrates that the GFP expression of cells is not altered by the SPIO labeling process. SPIO-labeled GFP cells can be visualized by MRI; therefore, GFP, a gene marker, was tracked indirectly with the SPIO-loaded cells using MRI. The technique holds promise for monitoring the temporal and spatial migration of cells with a gene marker and enhancing the understanding of cell- and gene-based therapeutic strategies. PMID:18956269

  9. Stereoscopic Feature Tracking System for Retrieving Velocity of Surface Waters

    Zuniga Zamalloa, C. C.; Landry, B. J.

    2017-12-01

    The present work is concerned with the surface velocity retrieval of flows using a stereoscopic setup and finding the correspondence in the images via feature tracking (FT). The feature tracking provides a key benefit of substantially reducing the level of user input. In contrast to other commonly used methods (e.g., normalized cross-correlation), FT does not require the user to prescribe interrogation window sizes and removes the need for masking when specularities are present. The results of the current FT methodology are comparable to those obtained via Large Scale Particle Image Velocimetry while requiring little to no user input which allowed for rapid, automated processing of imagery.

  10. Thyroid cell irradiation by radioiodines: a new Monte Carlo electron track-structure code

    Christophe Champion

    2007-09-01

    Full Text Available The most significant impact of the Chernobyl accident is the increased incidence of thyroid cancer among children who were exposed to short-lived radioiodines and 131-iodine. In order to accurately estimate the radiation dose provided by these radioiodines, it is necessary to know where iodine is incorporated. To do that, the distribution at the cellular level of newly organified iodine in the immature rat thyroid was performed using secondary ion mass microscopy (NanoSIMS50. Actual dosimetric models take only into account the averaged energy and range of beta particles of the radio-elements and may, therefore, imperfectly describe the real distribution of dose deposit at the microscopic level around the point sources. Our approach is radically different since based on a track-structure Monte Carlo code allowing following-up of electrons down to low energies (~ 10eV what permits a nanometric description of the irradiation physics. The numerical simulations were then performed by modelling the complete disintegrations of the short-lived iodine isotopes as well as of 131I in new born rat thyroids in order to take into account accurate histological and biological data for the thyroid gland.O impacto mais significante do acidente de Chernobyl é o crescimento da incidência de câncer de tireóide em crianças que foram expostas a radioiodos de vida curta e ao Iodo-131. Na estimativa precisa da dose de radiação fornecida por esses radioiodos, é necessário conhecer onde o iodo está incorporado. Para obtermos esse resultado, a distribuição em nível celular de iodo recentemente organificado na tireóde de ratos imaturos foi realizada usando microscopia de massa iônica secundária (NanoSIMS50. Modelos dosimétricos atuais consideram apenas a energia média das partículas beta dos radioelementos e pode, imperfeitamente descrever a distribuição real de dose ao nível microscópico em torno dos pontos pesquisados. Nossa abordagem

  11. Click chemistry-based tracking reveals putative cell wall-located auxin binding sites in expanding cells

    Mravec, J.; Kračun, S. K.; Zemlyanskaya, E.; Rydahl, M. G.; Guo, X.; Pičmanová, M.; Sørensen, K.; Růžička, Kamil; Willats, W.G.T.

    2017-01-01

    Roč. 7, NOV 22 (2017), č. článku 15988. ISSN 2045-2322 R&D Projects: GA MŠk(CZ) LQ1601 Institutional support: RVO:61389030 Keywords : MEMBRANE H+-ATPASE * BIOLOGICAL-ACTIVITY * AZIDO AUXINS Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 4.259, year: 2016

  12. Multiparticle imaging technique for two-phase fluid flows using pulsed laser speckle velocimetry. Final report, September 1988--November 1992

    Hassan, T.A.

    1992-12-01

    The practical use of Pulsed Laser Velocimetry (PLV) requires the use of fast, reliable computer-based methods for tracking numerous particles suspended in a fluid flow. Two methods for performing tracking are presented. One method tracks a particle through multiple sequential images (minimum of four required) by prediction and verification of particle displacement and direction. The other method, requiring only two sequential images uses a dynamic, binary, spatial, cross-correlation technique. The algorithms are tested on computer-generated synthetic data and experimental data which was obtained with traditional PLV methods. This allowed error analysis and testing of the algorithms on real engineering flows. A novel method is proposed which eliminates tedious, undersirable, manual, operator assistance in removing erroneous vectors. This method uses an iterative process involving an interpolated field produced from the most reliable vectors. Methods are developed to allow fast analysis and presentation of sets of PLV image data. Experimental investigation of a two-phase, horizontal, stratified, flow regime was performed to determine the interface drag force, and correspondingly, the drag coefficient. A horizontal, stratified flow test facility using water and air was constructed to allow interface shear measurements with PLV techniques. The experimentally obtained local drag measurements were compared with theoretical results given by conventional interfacial drag theory. Close agreement was shown when local conditions near the interface were similar to space-averaged conditions. However, theory based on macroscopic, space-averaged flow behavior was shown to give incorrect results if the local gas velocity near the interface as unstable, transient, and dissimilar from the average gas velocity through the test facility.

  13. Image-preprocessing method for near-wall particle image velocimetry (PIV) image interrogation with very large in-plane displacement

    Zhu, Yiding; Yuan, Huijing; Zhang, Chuanhong; Lee, Cunbiao

    2013-01-01

    Accurate particle image velocimetry (PIV) measurements very near the wall are still a great challenge. The problem is compounded by the very large in-plane displacement on PIV images commonly encountered in measurements in hypersonic boundary layers. An improved image-preprocessing method is presented in this paper which expands the traditional window deformation iterative multigrid scheme to PIV images with very large displacement. Before the interrogation, stationary artificial particles of uniform size are added homogeneously in the wall region. The mean squares of the intensities of signals in the flow and in the wall region are postulated to be equal when half the initial interrogation window overlaps the wall region. The initial estimation near the wall is then smoothed by data from both sides of the shear layer to reduce the large random uncertainties. Interrogations in the following iterative steps then converge to the correct results to provide accurate predictions for particle tracking velocimetries. Significant improvement is seen in Monte Carlo simulations and experimental tests. The algorithm successfully extracted the small flow structures of the second-mode wave in the hypersonic boundary layer from PIV images with low signal-noise-ratios when the traditional method was not successful. (paper)

  14. 2D ratiometric fluorescent pH sensor for tracking of cells proliferation and metabolism.

    Ma, Jun; Ding, Changqin; Zhou, Jie; Tian, Yang

    2015-08-15

    Extracellular pH plays a vital role no matter in physiological or pathological studies. In this work, a hydrogel, CD@Nile-FITC@Gel (Gel sensor), entrapping the ratiometric fluorescent probe CD@Nile-FITC was developed. The Gel sensor was successfully used for real-time extracellular pH monitoring. In the case of CD@Nile-FITC, pH-sensitive fluorescent dye fluorescein isothiocyanate (FITC) was chosen as the response signal for H(+) and Nile blue chloride (Nile) as the reference signal. The developed fluorescent probe exhibited high selectivity for pH over other metal ions and amino acids. Meanwhile, the carbon-dots-based inorganic-organic probe demonstrated excellent photostability against long-term light illumination. In order to study the extracellular pH change in processes of cell proliferation and metabolism, CD@Nile-FITC probe was entrapped in sodium alginate gel and consequently formed CD@Nile-FITC@Gel. The MTT assay showed low cytotoxicity of the Gel and the pH titration indicated that it could monitor the pH fluctuations linearly and rapidly within the pH range of 6.0-9.0, which is valuable for physiological pH determination. As expected, the real-time bioimaging of the probe was successfully achieved. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. T cell-based tracking of multidrug resistant tuberculosis infection after brief exposure.

    Richeldi, Luca; Ewer, Katie; Losi, Monica; Bergamini, Barbara M; Roversi, Pietro; Deeks, Jonathan; Fabbri, Leonardo M; Lalvani, Ajit

    2004-08-01

    Molecular epidemiology indicates significant transmission of Mycobacterium tuberculosis after casual contact with infectious tuberculosis cases. We investigated M. tuberculosis transmission after brief exposure using a T cell-based assay, the enzyme-linked-immunospot (ELISPOT) for IFN-gamma. After childbirth, a mother was diagnosed with sputum smear-positive multidrug-resistant tuberculosis. Forty-one neonates and 47 adults were present during her admission on the maternity unit; 11 weeks later, all underwent tuberculin skin testing (TST) and ELISPOT. We correlated test results with markers of exposure to the index case. The participants, who were asymptomatic and predominantly had no prior tuberculosis exposure, had 6.05 hours mean exposure (range: 0-65 hours) to the index case. Seventeen individuals, including two newborns, were ELISPOT-positive, and ELISPOT results correlated significantly with three of four predefined measures of tuberculosis exposure. For each hour sharing room air with the index case, the odds of a positive ELISPOT result increased by 1.05 (95% CI: 1.02-1.09, p = 0.003). Only four adults were TST-positive and TST results did not correlate with exposure. Thus, ELISPOT, but not TST, suggested quite extensive nosocomial transmission of multidrug-resistant M. tuberculosis after brief exposure. These results help to explain the apparent importance of casual contact for tuberculosis transmission, and may have implications for prevention.

  16. Implementation of a new blood cooler insert and tracking technology with educational initiatives and its effect on reducing red blood cell wastage.

    Fadeyi, Emmanuel A; Emery, Wanda; Simmons, Julie H; Jones, Mary Rose; Pomper, Gregory J

    2017-10-01

    The objective was to report a successful implementation of a blood cooler insert and tracking technology with educational initiatives and its effect on reducing red blood cell (RBC) wastage. The blood bank database was used to quantify and categorize total RBC units issued in blood coolers from January 2010 to December 2015 with and without the new inserts throughout the hospital. Radiofrequency identification tags were used with special software to monitor blood cooler tracking. An educational policy on how to handle the coolers was initiated. Data were gathered from the software that provided a real-time location monitoring of the blood coolers with inserts throughout the institution. The implementation of the blood cooler with inserts and tracking device reduced mean yearly RBC wastage by fourfold from 0.64% to 0.17% between 2010 and 2015. The conserved RBCs corresponded to a total cost savings of $167,844 during the 3-year postimplementation period. The implementation of new blood cooler inserts, tracking system, and educational initiatives substantially reduced the mean annual total RBC wastage. The cost to implement this initiative may be small if there is an existing institutional infrastructure to monitor and track hospital equipment into which the blood bank intervention can be adapted when compared to the cost of blood wastage. © 2017 AABB.

  17. Hierarchical Load Tracking Control of a Grid-Connected Solid Oxide Fuel Cell for Maximum Electrical Efficiency Operation

    Yonghui Li

    2015-03-01

    Full Text Available Based on the benchmark solid oxide fuel cell (SOFC dynamic model for power system studies and the analysis of the SOFC operating conditions, the nonlinear programming (NLP optimization method was used to determine the maximum electrical efficiency of the grid-connected SOFC subject to the constraints of fuel utilization factor, stack temperature and output active power. The optimal operating conditions of the grid-connected SOFC were obtained by solving the NLP problem considering the power consumed by the air compressor. With the optimal operating conditions of the SOFC for the maximum efficiency operation obtained at different active power output levels, a hierarchical load tracking control scheme for the grid-connected SOFC was proposed to realize the maximum electrical efficiency operation with the stack temperature bounded. The hierarchical control scheme consists of a fast active power control and a slower stack temperature control. The active power control was developed by using a decentralized control method. The efficiency of the proposed hierarchical control scheme was demonstrated by case studies using the benchmark SOFC dynamic model.

  18. Dual-wavelength OR-PAM with compressed sensing for cell tracking in a 3D cell culture system

    Huang, Rou-Xuan; Fu, Ying; Liu, Wang; Ma, Yu-Ting; Hsieh, Bao-Yu; Chen, Shu-Ching; Sun, Mingjian; Li, Pai-Chi

    2018-02-01

    Monitoring dynamic interactions of T cells migrating toward tumor is beneficial to understand how cancer immunotherapy works. Optical-resolution photoacoustic microscope (OR-PAM) can provide not only high spatial resolution but also deeper penetration than conventional optical microscopy. With the aid of exogenous contrast agents, the dual-wavelength OR-PAM can be applied to map the distribution of CD8+ cytotoxic T lymphocytes (CTLs) with gold nanospheres (AuNS) under 523nm laser irradiation and Hepta1-6 tumor spheres with indocyanine green (ICG) under 800nm irradiation. However, at 1K laser PRF, it takes approximately 20 minutes to obtain a full sample volume of 160 × 160 × 150 μm3 . To increase the imaging rate, we propose a random non-uniform sparse sampling mechanism to achieve fast sparse photoacoustic data acquisition. The image recovery process is formulated as a low-rank matrix recovery (LRMR) based on compressed sensing (CS) theory. We show that it could be stably recovered via nuclear-norm minimization optimization problem to maintain image quality from a significantly fewer measurement. In this study, we use the dual-wavelength OR-PAM with CS to visualize T cell trafficking in a 3D culture system with higher temporal resolution. Data acquisition time is reduced by 40% in such sample volume where sampling density is 0.5. The imaging system reveals the potential to understand the dynamic cellular process for preclinical screening of anti-cancer drugs.

  19. Convergence of lateral dynamic measurements in the plasma membrane of live cells from single particle tracking and STED-FCS

    Lagerholm, B Christoffer; Eggeling, Christian; Andrade, Débora M; Clausen, Mathias P

    2017-01-01

    Fluorescence correlation spectroscopy (FCS) in combination with the super-resolution imaging method STED (STED-FCS), and single-particle tracking (SPT) are able to directly probe the lateral dynamics of lipids and proteins in the plasma membrane of live cells at spatial scales much below the diffraction limit of conventional microscopy. However, a major disparity in interpretation of data from SPT and STED-FCS remains, namely the proposed existence of a very fast (unhindered) lateral diffusion coefficient, ⩾5 µ m 2 s −1 , in the plasma membrane of live cells at very short length scales, ≈⩽ 100 nm, and time scales, ≈1–10 ms. This fast diffusion coefficient has been advocated in several high-speed SPT studies, for lipids and membrane proteins alike, but the equivalent has not been detected in STED-FCS measurements. Resolving this ambiguity is important because the assessment of membrane dynamics currently relies heavily on SPT for the determination of heterogeneous diffusion. A possible systematic error in this approach would thus have vast implications in this field. To address this, we have re-visited the analysis procedure for SPT data with an emphasis on the measurement errors and the effect that these errors have on the measurement outputs. We subsequently demonstrate that STED-FCS and SPT data, following careful consideration of the experimental errors of the SPT data, converge to a common interpretation which for the case of a diffusing phospholipid analogue in the plasma membrane of live mouse embryo fibroblasts results in an unhindered, intra-compartment, diffusion coefficient of  ≈0.7–1.0 µ m 2 s −1 , and a compartment size of about 100–150 nm. (topical review)

  20. Effect of non-Poisson samples on turbulence spectra from laser velocimetry

    Sree, Dave; Kjelgaard, Scott O.; Sellers, William L., III

    1994-01-01

    Spectral analysis of laser velocimetry (LV) data plays an important role in characterizing a turbulent flow and in estimating the associated turbulence scales, which can be helpful in validating theoretical and numerical turbulence models. The determination of turbulence scales is critically dependent on the accuracy of the spectral estimates. Spectral estimations from 'individual realization' laser velocimetry data are typically based on the assumption of a Poisson sampling process. What this Note has demonstrated is that the sampling distribution must be considered before spectral estimates are used to infer turbulence scales.

  1. An innovative experimental setup for Large Scale Particle Image Velocimetry measurements in riverine environments

    Tauro, Flavia; Olivieri, Giorgio; Porfiri, Maurizio; Grimaldi, Salvatore

    2014-05-01

    Large Scale Particle Image Velocimetry (LSPIV) is a powerful methodology to nonintrusively monitor surface flows. Its use has been beneficial to the development of rating curves in riverine environments and to map geomorphic features in natural waterways. Typical LSPIV experimental setups rely on the use of mast-mounted cameras for the acquisition of natural stream reaches. Such cameras are installed on stream banks and are angled with respect to the water surface to capture large scale fields of view. Despite its promise and the simplicity of the setup, the practical implementation of LSPIV is affected by several challenges, including the acquisition of ground reference points for image calibration and time-consuming and highly user-assisted procedures to orthorectify images. In this work, we perform LSPIV studies on stream sections in the Aniene and Tiber basins, Italy. To alleviate the limitations of traditional LSPIV implementations, we propose an improved video acquisition setup comprising a telescopic, an inexpensive GoPro Hero 3 video camera, and a system of two lasers. The setup allows for maintaining the camera axis perpendicular to the water surface, thus mitigating uncertainties related to image orthorectification. Further, the mast encases a laser system for remote image calibration, thus allowing for nonintrusively calibrating videos without acquiring ground reference points. We conduct measurements on two different water bodies to outline the performance of the methodology in case of varying flow regimes, illumination conditions, and distribution of surface tracers. Specifically, the Aniene river is characterized by high surface flow velocity, the presence of abundant, homogeneously distributed ripples and water reflections, and a meagre number of buoyant tracers. On the other hand, the Tiber river presents lower surface flows, isolated reflections, and several floating objects. Videos are processed through image-based analyses to correct for lens

  2. Labeling Human Mesenchymal Stem Cells with Gold Nanocages for in vitro and in vivo Tracking by Two-Photon Microscopy and Photoacoustic Microscopy

    Zhang, Yu Shrike; Wang, Yu; Wang, Lidai; Wang, Yucai; Cai, Xin; Zhang, Chi; Wang, Lihong V.; Xia, Younan

    2013-01-01

    Stem cell tracking is a highly important subject. Current techniques based on nanoparticle-labeling, such as magnetic resonance imaging, fluorescence microscopy, and micro-computed tomography, are plagued by limitations including relatively low sensitivity or penetration depth, involvement of ionizing irradiation, and potential cytotoxicity of the nanoparticles. Here we introduce a new class of contrast agents based on gold nanocages (AuNCs) with hollow interiors and porous walls to label human mesenchymal stem cells (hMSCs) for both in vitro and in vivo tracking using two-photon microscopy and photoacoustic microscopy. As demonstrated by the viability assay, the AuNCs showed negligible cytotoxicity under a reasonable dose, and did not alter the differentiation potential of the hMSCs into desired lineages. We were able to image the cells labeled with AuNCs in vitro for at least 28 days in culture, as well as to track the cells that homed to the tumor region in nude mice in vivo. PMID:23946820

  3. Evaluation of a BGO-Based PET System for Single-Cell Tracking Performance by Simulation and Phantom Studies

    Yu Ouyang PhD

    2016-05-01

    Full Text Available A recent method based on positron emission was reported for tracking moving point sources using the Inveon PET system. However, the effect of scanner background noise was not further explored. Here, we evaluate tracking with the Genisys4, a bismuth germanate-based PET system, which has no significant intrinsic background and may be better suited to tracking lower and/or faster activity sources. Position-dependent sensitivity of the Genisys4 was simulated in Geant4 Application for Tomographic Emission (GATE using a static 18F point source. Trajectories of helically moving point sources with varying activity and rotation speed were reconstructed from list-mode data as described previously. Simulations showed that the Inveon’s ability to track sources within 2 mm of localization error is limited to objects with a velocity-to-activity ratio < 0.13 mm/decay, compared to < 0.29 mm/decay for the Genisys4. Tracking with the Genisys4 was then validated using a physical phantom of helically moving [18F] fluorodeoxyglucose-in-oil droplets (< 0.24 mm diameter, 139-296 Bq, yielding < 1 mm localization error under the tested conditions, with good agreement between simulated sensitivity and measured activity (Pearson correlation R = .64, P << .05 in a representative example. We have investigated the tracking performance with the Genisys4, and results suggest the feasibility of tracking low activity, point source-like objects with this system.

  4. Particle tracking

    Mais, H.; Ripken, G.; Wrulich, A.; Schmidt, F.

    1986-02-01

    After a brief description of typical applications of particle tracking in storage rings and after a short discussion of some limitations and problems related with tracking we summarize some concepts and methods developed in the qualitative theory of dynamical systems. We show how these concepts can be applied to the proton ring HERA. (orig.)

  5. Timber tracking

    Düdder, Boris; Ross, Omry

    2017-01-01

    Managing and verifying forest products in a value chain is often reliant on easily manipulated document or digital tracking methods - Chain of Custody Systems. We aim to create a new means of tracking timber by developing a tamper proof digital system based on Blockchain technology. Blockchain...

  6. QUANTITATIVE FLOW-ANALYSIS AROUND AQUATIC ANIMALS USING LASER SHEET PARTICLE IMAGE VELOCIMETRY

    STAMHUIS, EJ; VIDELER, JJ

    Two alternative particle image velocimetry (PIV) methods have been developed, applying laser light sheet illumination of particle-seeded flows around marine organisms, Successive video images, recorded perpendicular to a light sheet parallel to the main stream, were digitized and processed to map

  7. Analysis of propeller-induced ground vortices by particle image velocimetry

    Yang, Y.; Sciacchitano, A.; Veldhuis, L.L.M.; Eitelberg, G.

    2017-01-01

    Abstract: The interaction between a propeller and its self-induced vortices originating on the ground is investigated in a scaled experiment. The velocity distribution in the flow field in two different planes containing the self-induced vortices is measured by particle image velocimetry (PIV).

  8. Endovascular Device Testing with Particle Image Velocimetry Enhances Undergraduate Biomedical Engineering Education

    Nair, Priya; Ankeny, Casey J.; Ryan, Justin; Okcay, Murat; Frakes, David H.

    2016-01-01

    We investigated the use of a new system, HemoFlow™, which utilizes state of the art technologies such as particle image velocimetry to test endovascular devices as part of an undergraduate biomedical engineering curriculum. Students deployed an endovascular stent into an anatomical model of a cerebral aneurysm and measured intra-aneurysmal flow…

  9. Three-dimensional particle image velocimetry in a generic can-type gas turbine combustor

    Meyers, BC

    2009-09-01

    Full Text Available The three-dimensional flow field inside a generic can-type, forward flow, experimental combustor was measured. A stereoscopic Particle Image Velocimetry (PIV) system was used to obtain the flow field of the combustor in the non-reacting condition...

  10. Rainbow particle imaging velocimetry for dense 3D fluid velocity imaging

    Xiong, Jinhui

    2017-07-21

    Despite significant recent progress, dense, time-resolved imaging of complex, non-stationary 3D flow velocities remains an elusive goal. In this work we tackle this problem by extending an established 2D method, Particle Imaging Velocimetry, to three dimensions by encoding depth into color. The encoding is achieved by illuminating the flow volume with a continuum of light planes (a

  11. Particle imaging velocimetry experiments and lattice-Boltzmann simulations on a single sphere settling under gravity

    Ten Cate, A.; Nieuwstad, C.H.; Derksen, J.J.; Van den Akker, H.E.A.

    2002-01-01

    A comparison is made between experiments and simulations on a single sphere settling in silicon oil in a box. Cross-correlation particle imaging velocimetry measurements were carried out at particle Reynolds numbers ranging from 1.5 to 31.9. The particle Stokes number varied from 0.2 to 4 and at

  12. Particle image velocimetry measurements and numerical modeling of a saline density current

    Gerber, G

    2011-03-01

    Full Text Available Particle image velocimetry scalar measurements were carried out on the body of a stably stratified density current with an inlet Reynolds number of 2,300 and bulk Richardson number of 0.1. These measurements allowed the mass and momentum transport...

  13. Three-component particle image velocimetry in a generic can-type gas turbine combustor

    Meyers, Bronwyn C

    2012-11-01

    Full Text Available -1 Proceedings of the Institution of Mechanical Engineers, Part A: Journal of Power and Energy November 2012/ Vol. 226(7) Three-componentParticle Image Velocimetry in a Generic Can-type Gas Turbine Combustor B C Meyers 1, 2* , G C Snedden 1 , J P...

  14. Fluid Flow Characterization of High Turbulent Intensity Compressible Flow Using Particle Image Velocimetry

    2015-08-01

    completed in order to begin further experimentation. A 10 kHz Time Resolved Particle Image Velocimetry (TR-PIV) system and a 3 kHz Planer Laser ...9 2.3.2 Planar Laser Induced Fluorescence (PLIF...35 Figure 4.4: Solenoid valve (a), proportional control valve (b) and flowmeter (c) ...................................... 36 Figure 4.5

  15. Drag coefficient accuracy improvement by means of particle image velocimetry for a transonic NACA0012 airfoil

    Ragni, D; Van Oudheusden, B W; Scarano, F

    2011-01-01

    A method to improve the reliability of the drag coefficient computation by means of particle image velocimetry measurements is made using experimental data acquired on a NACA0012 airfoil tested in the transonic regime, using the combination of a variable pulse separation with a new high-order Poisson spectral pressure reconstruction algorithm. (technical design note)

  16. Application of stereoscopic particle image velocimetry to studies of transport in a dusty (complex) plasma

    Thomas, Edward Jr.; Williams, Jeremiah D.; Silver, Jennifer

    2004-01-01

    Over the past 5 years, two-dimensional particle image velocimetry (PIV) techniques [E. Thomas, Jr., Phys. Plasmas 6, 2672 (1999)] have been used to obtain detailed measurements of microparticle transport in dusty plasmas. This Letter reports on an extension of these techniques to a three-dimensional velocity vector measurement approach using stereoscopic PIV. Initial measurements using the stereoscopic PIV diagnostic are presented

  17. Basics and principles of particle image velocimetry (PIV) for mapping biogenic and biologically relevant flows

    Stamhuis, Eize J.

    2006-01-01

    Particle image velocimetry (PIV) has proven to be a very useful technique in mapping animal-generated flows or flow patterns relevant to biota. Here, theoretical background is provided and experimental details of 2-dimensional digital PIV are explained for mapping flow produced by or relevant to

  18. The application of particle image velocimetry for the analysis of high-speed craft hydrodynamics

    Jacobi, G.; Thill, C.H.; Huijsmans, R.H.M.; Huijsmans, R.H.M.

    2016-01-01

    The particle image velocimetry (PIV) technique has become a reliable method for capturing the velocity field and its derivatives, even in complex flows and is now also widely used for validation of numerical codes. As the imaging system is sensitive to vibrations, the application in environments

  19. PIV as a method for quantifying root cell growth and particle displacement in confocal images.

    Bengough, A Glyn; Hans, Joachim; Bransby, M Fraser; Valentine, Tracy A

    2010-01-01

    Particle image velocimetry (PIV) quantifies displacement of patches of pixels between successive images. We evaluated PIV as a tool for microscopists by measuring displacements of cells and of a surrounding granular medium in confocal laser scanning microscopy images of Arabidopsis thaliana roots labeled with cell-membrane targeted green fluorescent protein. Excellent accuracy (e.g., displacement standard deviation PIV-predicted and actual displacements (r(2) > 0.83). Root mean squared error for these distorted images was 0.4-1.1 pixels, increasing at higher magnification factors. Cell growth and rhizosphere deformation were tracked with good temporal (e.g., 1-min interval) and spatial resolution, with PIV patches located on recognizable cell features being tracked more successfully. Appropriate choice of GFP-label was important to decrease small-scale biological noise due to intracellular motion. PIV of roots grown in stiff 2% versus 0.7% agar showed patterns of cell expansion consistent with physically impeded roots of other species. Roots in glass ballotini underwent rapid changes in growth direction on a timescale of minutes, associated with localized arching of ballotini. By tracking cell vertices, we monitored automatically cell length, width, and area every minute for 0.5 h for cells in different stages of development. In conclusion, PIV measured displacements successfully in images of living root cells and the external granular medium, revealing much potential for use by microscopists. (c) 2009 Wiley-Liss, Inc.

  20. Extracellular NGFR Spacers Allow Efficient Tracking and Enrichment of Fully Functional CAR-T Cells Co-Expressing a Suicide Gene.

    Casucci, Monica; Falcone, Laura; Camisa, Barbara; Norelli, Margherita; Porcellini, Simona; Stornaiuolo, Anna; Ciceri, Fabio; Traversari, Catia; Bordignon, Claudio; Bonini, Chiara; Bondanza, Attilio

    2018-01-01

    Chimeric antigen receptor (CAR)-T cell immunotherapy is at the forefront of innovative cancer therapeutics. However, lack of standardization of cellular products within the same clinical trial and lack of harmonization between different trials have hindered the clear identification of efficacy and safety determinants that should be unveiled in order to advance the field. With the aim of facilitating the isolation and in vivo tracking of CAR-T cells, we here propose the inclusion within the CAR molecule of a novel extracellular spacer based on the low-affinity nerve-growth-factor receptor (NGFR). We screened four different spacer designs using as target antigen the CD44 isoform variant 6 (CD44v6). We successfully generated NGFR-spaced CD44v6 CAR-T cells that could be efficiently enriched with clinical-grade immuno-magnetic beads without negative consequences on subsequent expansion, immuno-phenotype, in vitro antitumor reactivity, and conditional ablation when co-expressing a suicide gene. Most importantly, these cells could be tracked with anti-NGFR monoclonal antibodies in NSG mice, where they expanded, persisted, and exerted potent antitumor effects against both high leukemia and myeloma burdens. Similar results were obtained with NGFR-enriched CAR-T cells specific for CD19 or CEA, suggesting the universality of this strategy. In conclusion, we have demonstrated that the incorporation of the NGFR marker gene within the CAR sequence allows for a single molecule to simultaneously work as a therapeutic and selection/tracking gene. Looking ahead, NGFR spacer enrichment might allow good manufacturing procedures-manufacturing of standardized CAR-T cell products with high therapeutic potential, which could be harmonized in different clinical trials and used in combination with a suicide gene for future application in the allogeneic setting.

  1. Extracellular NGFR Spacers Allow Efficient Tracking and Enrichment of Fully Functional CAR-T Cells Co-Expressing a Suicide Gene

    Monica Casucci

    2018-03-01

    Full Text Available Chimeric antigen receptor (CAR-T cell immunotherapy is at the forefront of innovative cancer therapeutics. However, lack of standardization of cellular products within the same clinical trial and lack of harmonization between different trials have hindered the clear identification of efficacy and safety determinants that should be unveiled in order to advance the field. With the aim of facilitating the isolation and in vivo tracking of CAR-T cells, we here propose the inclusion within the CAR molecule of a novel extracellular spacer based on the low-affinity nerve-growth-factor receptor (NGFR. We screened four different spacer designs using as target antigen the CD44 isoform variant 6 (CD44v6. We successfully generated NGFR-spaced CD44v6 CAR-T cells that could be efficiently enriched with clinical-grade immuno-magnetic beads without negative consequences on subsequent expansion, immuno-phenotype, in vitro antitumor reactivity, and conditional ablation when co-expressing a suicide gene. Most importantly, these cells could be tracked with anti-NGFR monoclonal antibodies in NSG mice, where they expanded, persisted, and exerted potent antitumor effects against both high leukemia and myeloma burdens. Similar results were obtained with NGFR-enriched CAR-T cells specific for CD19 or CEA, suggesting the universality of this strategy. In conclusion, we have demonstrated that the incorporation of the NGFR marker gene within the CAR sequence allows for a single molecule to simultaneously work as a therapeutic and selection/tracking gene. Looking ahead, NGFR spacer enrichment might allow good manufacturing procedures-manufacturing of standardized CAR-T cell products with high therapeutic potential, which could be harmonized in different clinical trials and used in combination with a suicide gene for future application in the allogeneic setting.

  2. Extracellular NGFR Spacers Allow Efficient Tracking and Enrichment of Fully Functional CAR-T Cells Co-Expressing a Suicide Gene

    Casucci, Monica; Falcone, Laura; Camisa, Barbara; Norelli, Margherita; Porcellini, Simona; Stornaiuolo, Anna; Ciceri, Fabio; Traversari, Catia; Bordignon, Claudio; Bonini, Chiara; Bondanza, Attilio

    2018-01-01

    Chimeric antigen receptor (CAR)-T cell immunotherapy is at the forefront of innovative cancer therapeutics. However, lack of standardization of cellular products within the same clinical trial and lack of harmonization between different trials have hindered the clear identification of efficacy and safety determinants that should be unveiled in order to advance the field. With the aim of facilitating the isolation and in vivo tracking of CAR-T cells, we here propose the inclusion within the CAR molecule of a novel extracellular spacer based on the low-affinity nerve-growth-factor receptor (NGFR). We screened four different spacer designs using as target antigen the CD44 isoform variant 6 (CD44v6). We successfully generated NGFR-spaced CD44v6 CAR-T cells that could be efficiently enriched with clinical-grade immuno-magnetic beads without negative consequences on subsequent expansion, immuno-phenotype, in vitro antitumor reactivity, and conditional ablation when co-expressing a suicide gene. Most importantly, these cells could be tracked with anti-NGFR monoclonal antibodies in NSG mice, where they expanded, persisted, and exerted potent antitumor effects against both high leukemia and myeloma burdens. Similar results were obtained with NGFR-enriched CAR-T cells specific for CD19 or CEA, suggesting the universality of this strategy. In conclusion, we have demonstrated that the incorporation of the NGFR marker gene within the CAR sequence allows for a single molecule to simultaneously work as a therapeutic and selection/tracking gene. Looking ahead, NGFR spacer enrichment might allow good manufacturing procedures-manufacturing of standardized CAR-T cell products with high therapeutic potential, which could be harmonized in different clinical trials and used in combination with a suicide gene for future application in the allogeneic setting. PMID:29619024

  3. Aldehyde dehydrogenase 1 is a marker for normal and malignant human colonic stem cells (SC) and tracks SC overpopulation during colon tumorigenesis.

    Huang, Emina H; Hynes, Mark J; Zhang, Tao; Ginestier, Christophe; Dontu, Gabriela; Appelman, Henry; Fields, Jeremy Z; Wicha, Max S; Boman, Bruce M

    2009-04-15

    Although the concept that cancers originate from stem cells (SC) is becoming scientifically accepted, mechanisms by which SC contribute to tumor initiation and progression are largely unknown. For colorectal cancer (CRC), investigation of this problem has been hindered by a paucity of specific markers for identification and isolation of SC from normal and malignant colon. Accordingly, aldehyde dehydrogenase 1 (ALDH1) was investigated as a possible marker for identifying colonic SC and for tracking them during cancer progression. Immunostaining showed that ALDH1(+) cells are sparse and limited to the normal crypt bottom, where SCs reside. During progression from normal epithelium to mutant (APC) epithelium to adenoma, ALDH1(+) cells increased in number and became distributed farther up the crypt. CD133(+) and CD44(+) cells, which are more numerous and broadly distributed in normal crypts, showed similar changes during tumorigenesis. Flow cytometric isolation of cancer cells based on enzymatic activity of ALDH (Aldefluor assay) and implantation of these cells in nonobese diabetic-severe combined immunodeficient mice (a) generated xenograft tumors (Aldefluor(-) cells did not), (b) generated them after implanting as few as 25 cells, and (c) generated them dose dependently. Further isolation of cancer cells using a second marker (CD44(+) or CD133(+) serially) only modestly increased enrichment based on tumor-initiating ability. Thus, ALDH1 seems to be a specific marker for identifying, isolating, and tracking human colonic SC during CRC development. These findings also support our original hypothesis, derived previously from mathematical modeling of crypt dynamics, that progressive colonic SC overpopulation occurs during colon tumorigenesis and drives CRC development.

  4. Why tracks

    Burchart, J.; Kral, J.

    1979-01-01

    A comparison is made of two methods of determining the age of rocks, ie., the krypton-argon method and the fission tracks method. The former method is more accurate but is dependent on the temperature and on the grain size of the investigated rocks (apatites, biotites, muscovites). As for the method of fission tracks, the determination is not dependent on grain size. This method allows dating and the determination of uranium concentration and distribution in rocks. (H.S.)

  5. Near-ground tornado-like vortex structure resolved by particle image velocimetry (PIV)

    Zhang, Wei [Iowa State University, Aerospace Engineering Department, Ames, IA (United States); University of Minnesota, Saint Anthony Falls Laboratory, Minneapolis, MN (United States); Sarkar, Partha P. [Iowa State University, Aerospace Engineering Department, Ames, IA (United States)

    2012-02-15

    The near-ground flow structure of tornadoes is of utmost interest because it determines how and to what extent civil structures could get damaged in tornado events. We simulated tornado-like vortex flow at the swirl ratios of S = 0.03-0.3 (vane angle {theta}{sub v} = 15 -60 ), using a laboratory tornado simulator and investigated the near-ground-vortex structure by particle imaging velocimetry. Complicated near-ground flow was measured in two orthogonal views: horizontal planes at various elevations (z = 11, 26 and 53 mm above the ground) and the meridian plane. We observed two distinct vortex structures: a single-celled vortex at the lowest swirl ratio (S = 0.03, {theta}{sub v} = 15 ) and multiple suction vortices rotating around the primary vortex (two-celled vortex) at higher swirl ratios (S = 0.1-0.3, {theta}{sub v} = 30 -60 ). We quantified the effects of vortex wandering on the mean flow and found that vortex wandering was important and should be taken into account in the low swirl ratio case. The tangential velocity, as the dominant velocity component, has the peak value about three times that of the maximum radial velocity regardless of the swirl ratio. The maximum velocity variance is about twice at the high swirl ratio ({theta}{sub v} = 45 ) that at the low swirl ratio ({theta}{sub v} = 15 ), which is contributed significantly by the multiple small-scale secondary vortices. Here, the results show that not only the intensified mean flow but greatly enhanced turbulence occurs near the surface in the tornado-like vortex flow. The intensified mean flow and enhanced turbulence at the ground level, correlated with the ground-vortex interaction, may cause dramatic damage of the civil structures in tornadoes. This work provides detailed characterization of the tornado-like vortex structure, which has not been fully revealed in previous field studies and laboratory simulations. It would be helpful in improving the understanding of the interaction between the

  6. Optimal Load-Tracking Operation of Grid-Connected Solid Oxide Fuel Cells through Set Point Scheduling and Combined L1-MPC Control

    Siwei Han

    2018-03-01

    Full Text Available An optimal load-tracking operation strategy for a grid-connected tubular solid oxide fuel cell (SOFC is studied based on the steady-state analysis of the system thermodynamics and electrochemistry. Control of the SOFC is achieved by a two-level hierarchical control system. In the upper level, optimal setpoints of output voltage and the current corresponding to unit load demand is obtained through a nonlinear optimization by minimizing the SOFC’s internal power waste. In the lower level, a combined L1-MPC control strategy is designed to achieve fast set point tracking under system nonlinearities, while maintaining a constant fuel utilization factor. To prevent fuel starvation during the transient state resulting from the output power surging, a fuel flow constraint is imposed on the MPC with direct electron balance calculation. The proposed control schemes are testified on the grid-connected SOFC model.

  7. Track detection on the cells exposed to high Linear Energy Transfer heavy-ions by Cr-39 plastic and terminal deoxynucleotidyl transferase(Td T)

    Mehnati, P.; Keshtkar, A.; Mesbahi, A.; Sasaki, H.

    2006-01-01

    The fatal effect of ionizing radiation on cells depends on Linear Energy Transfer level. The distribution of ionizing radiation is sparse and homogeneous for low Linear Energy Transfer radiations such as X or y, but it is dense and concentrated for high Linear Energy Transfer radiation such as heavy-ions radiation. Materials and Methods: Chinese hamster ovary cells (CHO-K1) were exposed to 4 Gy Fe-ion 2000 keV/μm. The Cr-39 is a special and sensitive plastic used to verify exact position of heavy-ions traversal. Terminal deoxynucleotidyl transferase is an enzyme labeled with [3 H ] d ATP for detection of cellular DNA damage by autoradiography assay. Results: The track of heavy ions traversals presented by pit size was almost similar for all different doses of radiation. No pits to show the track of traversal were found in 20% of the cell nuclei of the irradiation. Apparently these fractions of cells wave not hit by heavy ions. Conclusion: This study indicated the possible usefulness of both the Cr-39 plastics and DNA labeling with Terminal deoxynucleotidyl transferase method for evaluating the biological effect of heavy-ions in comparison with low Linear Energy Transfer ionizing radiation

  8. Magnetic resonance cell-tracking studies: spectrophotometry-based method for the quantification of cellular iron content after loading with superparamagnetic iron oxide nanoparticles.

    Böhm, Ingrid

    2011-08-01

    The purpose of this article is to present a user-friendly tool for quantifying the iron content of superparamagnetic labeled cells before cell tracking by magnetic resonance imaging (MRI). Iron quantification was evaluated by using Prussian blue staining and spectrophotometry. White blood cells were labeled with superparamagnetic iron oxide (SPIO) nanoparticles. Labeling was confirmed by light microscopy. Subsequently, the cells were embedded in a phantom and scanned on a 3 T magnetic resonance tomography (MRT) whole-body system. Mean peak wavelengths λ(peak) was determined at A(720 nm) (range 719-722 nm). Linearity was proven for the measuring range 0.5 to 10 μg Fe/mL (r  =  .9958; p  =  2.2 × 10(-12)). The limit of detection was 0.01 μg Fe/mL (0.1785 mM), and the limit of quantification was 0.04 μg Fe/mL (0.714 mM). Accuracy was demonstrated by comparison with atomic absorption spectrometry. Precision and robustness were also proven. On T(2)-weighted images, signal intensity varied according to the iron concentration of SPIO-labeled cells. Absorption spectrophotometry is both a highly sensitive and user-friendly technique that is feasible for quantifying the iron content of magnetically labeled cells. The presented data suggest that spectrophotometry is a promising tool for promoting the implementation of magnetic resonance-based cell tracking in routine clinical applications (from bench to bedside).

  9. Magnetic Resonance Cell-Tracking Studies: Spectrophotometry-Based Method for the Quantification of Cellular Iron Content after Loading with Superparamagnetic Iron Oxide Nanoparticles

    Ingrid Böhm

    2011-07-01

    Full Text Available The purpose of this article is to present a user-friendly tool for quantifying the iron content of superparamagnetic labeled cells before cell tracking by magnetic resonance imaging (MRI. Iron quantification was evaluated by using Prussian blue staining and spectrophotometry. White blood cells were labeled with superparamagnetic iron oxide (SPIO nanoparticles. Labeling was confirmed by light microscopy. Subsequently, the cells were embedded in a phantom and scanned on a 3 T magnetic resonance tomography (MRT whole-body system. Mean peak wavelengths Λpeak was determined at A720nm (range 719–722 nm. Linearity was proven for the measuring range 0.5 to 10 μg Fe/mL (r = .9958; p = 2.2 × 10−12. The limit of detection was 0.01 μg Fe/mL (0.1785 mM, and the limit of quantification was 0.04 μg Fe/mL (0.714 mM. Accuracy was demonstrated by comparison with atomic absorption spectrometry. Precision and robustness were also proven. On T2-weighted images, signal intensity varied according to the iron concentration of SPIO-labeled cells. Absorption spectrophotometry is both a highly sensitive and user-friendly technique that is feasible for quantifying the iron content of magnetically labeled cells. The presented data suggest that spectrophotometry is a promising tool for promoting the implementation of magnetic resonance-based cell tracking in routine clinical applications (from bench to bedside.

  10. A parallel algorithm for 3D particle tracking and Lagrangian trajectory reconstruction

    Barker, Douglas; Zhang, Yuanhui; Lifflander, Jonathan; Arya, Anshu

    2012-01-01

    Particle-tracking methods are widely used in fluid mechanics and multi-target tracking research because of their unique ability to reconstruct long trajectories with high spatial and temporal resolution. Researchers have recently demonstrated 3D tracking of several objects in real time, but as the number of objects is increased, real-time tracking becomes impossible due to data transfer and processing bottlenecks. This problem may be solved by using parallel processing. In this paper, a parallel-processing framework has been developed based on frame decomposition and is programmed using the asynchronous object-oriented Charm++ paradigm. This framework can be a key step in achieving a scalable Lagrangian measurement system for particle-tracking velocimetry and may lead to real-time measurement capabilities. The parallel tracking algorithm was evaluated with three data sets including the particle image velocimetry standard 3D images data set #352, a uniform data set for optimal parallel performance and a computational-fluid-dynamics-generated non-uniform data set to test trajectory reconstruction accuracy, consistency with the sequential version and scalability to more than 500 processors. The algorithm showed strong scaling up to 512 processors and no inherent limits of scalability were seen. Ultimately, up to a 200-fold speedup is observed compared to the serial algorithm when 256 processors were used. The parallel algorithm is adaptable and could be easily modified to use any sequential tracking algorithm, which inputs frames of 3D particle location data and outputs particle trajectories

  11. Combined Lorentz force and ultrasound Doppler velocimetry in a vertical convection liquid metal flow

    Zürner, Till; Vogt, Tobias; Resagk, Christian; Eckert, Sven; Schumacher, Jörg

    2017-11-01

    We report experimental studies on turbulent vertical convection flow in the liquid metal alloy gallium-indium-tin. Flow measurements were conducted by a combined use of local Lorentz force velocimetry (LLFV) and ultrasound Doppler velocimetry (UDV). It is known that the forced convection flow in a duct generates a force on the LLFV magnet system, that grows proportional to the flow velocity. We show that for the slower flow of natural convection LLFV retains this linear dependence in the range of micronewtons. Furthermore experimental results on the scaling of heat and momentum transport with the thermal driving are presented. The results cover a range of Rayleigh numbers 3 ×105 Deutsche Forschungsgemeinschaft under Grant No. GRK 1567.

  12. Pulsed operation of high-power light emitting diodes for imaging flow velocimetry

    Willert, C; Klinner, J; Moessner, S; Stasicki, B

    2010-01-01

    High-powered light emitting diodes (LED) are investigated for possible uses as light sources in flow diagnostics, in particular, as an alternative to laser-based illumination in particle imaging flow velocimetry in side-scatter imaging arrangements. Recent developments in solid state illumination resulted in mass-produced LEDs that provide average radiant power in excess of 10 W. By operating these LEDs with short duration, pulsed currents that are considerably beyond their continuous current damage threshold, light pulses can be generated that are sufficient to illuminate and image micron-sized particles in flow velocimetry. Time-resolved PIV measurements in water at a framing rate of 2kHz are presented. The feasibility of LED-based PIV measurements in air is also demonstrated

  13. Assessment of Doppler velocimetry versus nonstress test in antepartum surveillance of high risk pregnancy

    Nishi Choudhury; Barun Kumar Sharma; Bikram Kishor Kanungo; Ruby Yadav; Hafizur Rahman

    2017-01-01

    Background: High risk pregnancies increase the maternal and fetal morbidity and mortality; and there is a need for appropriate investigation which can diagnose it early and predicts the morbidity and mortality. The objectives of this study were to compare the efficacy of Doppler velocimetry studies and NST in predicting fetal compromise in utero and compare their ability in predicting the perinatal outcome in cases of high risk pregnancies. Methods: It was a prospective cross-sectional ho...

  14. Abnormal Doppler flow velocimetry in the growth restricted foetus as a predictor for necrotising enterocolitis.

    Bhatt A

    2002-07-01

    Full Text Available BACKGROUND: Obstetric decision- making for the growth restricted foetus has to take into consideration the benefits and risks of waiting for pulmonary maturity and continued exposure to hostile intra-uterine environment. Necrotising Enterocolitis (NEC results from continued exposure to hostile environment and is an important cause of poor neonatal outcome. AIMS: To evaluate the predictive value of abnormal Doppler flow velocimetry of the foetal umbilical artery for NEC and neonatal mortality. SETTINGS AND DESIGN: A retrospective study carried out at a tertiary care centre for obstetric and neonatal care. MATERIALS AND METHOD: Seventy-seven neonates with birth weight less than 2000 gm, born over a period of 18 months were studied. These pregnancies were identified as having growth abnormalities of the foetus. Besides other tests of foetal well-being, they were also subjected to Doppler flow velocimetry of the foeto-placental vasculature. Obstetric outcome was evaluated with reference to period of gestation and route of delivery. The neonatal outcome was reviewed with reference to birth weight, Apgar scores and evidence of NEC. STATISTICAL ANALYSIS USED: Chi square test. RESULTS: In the group of patients with Absent or Reverse End Diastolic Frequencies (A/R EDF in the umbilical arteries, positive predictive value for NEC was 52.6%, (RR 30.2; OR 264. The mortality from NEC was 50%. When umbilical artery velocimetry did not show A/REDF, there were no cases of NEC or mortality. Abnormal umbilical or uterine artery flow increased the rate of caesarean section to 62.5% as compared to 17.6% in cases where umbilical artery flow was normal. CONCLUSION: In antenatally identified pregnancies at risk for foetal growth restriction, abnormal Doppler velocimetry in the form of A/REDF in the umbilical arteries is a useful guide to predict NEC and mortality in the early neonatal period.

  15. Tomographic Particle Image Velocimetry using Pulsed, High Power LED Volume Illumination

    Buchmann, N. A.; Willert, C.; Soria, J.

    2011-01-01

    This paper investigates the use of high-power light emitting diode (LED) illumination in Particle Image Velocimetry (PIV) as an alternative to traditional laser-based illumination. The solid-state LED devices can provide averaged radiant power in excess of 10W and by operating the LEDs with short current pulses, considerably higher than in continuous operation, light pulses of sufficient energy suitable for imaging micron-sized particles can be generated. The feasibility of this LED-based ill...

  16. A novel approach to tracking antigen-experienced CD4 T cells into functional compartments via tandem deep and shallow TCR clonotyping.

    Estorninho, Megan; Gibson, Vivienne B; Kronenberg-Versteeg, Deborah; Liu, Yuk-Fun; Ni, Chester; Cerosaletti, Karen; Peakman, Mark

    2013-12-01

    Extensive diversity in the human repertoire of TCRs for Ag is both a cornerstone of effective adaptive immunity that enables host protection against a multiplicity of pathogens and a weakness that gives rise to potential pathological self-reactivity. The complexity arising from diversity makes detection and tracking of single Ag-specific CD4 T cells (ASTs) involved in these immune responses challenging. We report a tandem, multistep process to quantify rare TCRβ-chain variable sequences of ASTs in large polyclonal populations. The approach combines deep high-throughput sequencing (HTS) within functional CD4 T cell compartments, such as naive/memory cells, with shallow, multiple identifier-based HTS of ASTs identified by activation marker upregulation after short-term Ag stimulation in vitro. We find that clonotypes recognizing HLA class II-restricted epitopes of both pathogen-derived Ags and self-Ags are oligoclonal and typically private. Clonotype tracking within an individual reveals private AST clonotypes resident in the memory population, as would be expected, representing clonal expansions (identical nucleotide sequence; "ultraprivate"). Other AST clonotypes share CDR3β amino acid sequences through convergent recombination and are found in memory populations of multiple individuals. Tandem HTS-based clonotyping will facilitate studying AST dynamics, epitope spreading, and repertoire changes that arise postvaccination and following Ag-specific immunotherapies for cancer and autoimmune disease.

  17. Non-invasive tracking of human haemopoietic CD34{sup +} stem cells in vivo in immunodeficient mice by using magnetic resonance imaging

    Niemeyer, Markus; Jacobs, Volker R.; Timmer, Sebastian; Kiechle, Marion [Technische Universitaet Muenchen, Department of Gynaecology, Klinikum rechts der Isar, Munich (Germany); Oostendorp, Robert A.J.; Hippauf, Sandra; Bekker-Ruz, Viktoria [Technische Universitaet Muenchen, Department of Oncology, Klinikum rechts der Isar, Munich (Germany); Kremer, Markus [Technische Universitaet Muenchen, Department of Pathology, Klinikum rechts der Isar, Munich (Germany); Baurecht, Hansjoerg [Technische Universitaet Muenchen, Department of Statistics, Klinikum rechts der Isar, Institute for Medical Statistics and Epidemiology, Munich (Germany); Ludwig, Georg; Rummeny, Ernst J. [Technische Universitaet Muenchen, Department of Radiology, Klinikum rechts der Isar, Munich (Germany); Piontek, Guido [Technische Universitaet Muenchen, Department of Neuropathology, Munich (Germany); Beer, Ambros J. [Technische Universitaet Muenchen, Department of Radiology, Klinikum rechts der Isar, Munich (Germany); Technische Universitaet Muenchen, Department of Nuclear Medicine, Munich (Germany)

    2010-09-15

    To assess migration of CD34{sup +} human stem cells to the bone marrow of athymic mice by using magnetic resonance (MR) imaging and Resovist, a contrast agent containing superparamagnetic iron oxide (SPIO) particles. All animal and human procedures were approved by our institution's ethics committee, and women had given consent to donate umbilical cord blood (UCB). Balb/c-AnN Foxn1{sup nu}/Crl mice received intravenous injection of 1 x 10{sup 6} (n = 3), 5 x 10{sup 6} (n = 3) or 1 x 10{sup 7} (n = 3) human Resovist-labelled CD34{sup +} cells; control mice received Resovist (n = 3). MR imaging was performed before, 2 and 24 h after transplantation. Signal intensities of liver, muscle and bone marrow were measured and analysed by ANOVA and post hoc Student's t tests. MR imaging data were verified by histological and immunological detection of both human cell surface markers and carboxydextran-coating of the contrast agent. CD34{sup +} cells were efficiently labelled by Resovist without impairment of functionality. Twenty-four hours after administration of labelled cells, MR imaging revealed a significant signal decline in the bone marrow, and histological and immunological analyses confirmed the presence of transplanted human CD34{sup +} cells. Intravenously administered Resovist-labelled CD34{sup +} cells home to bone marrow of mice. Homing can be tracked in vivo by using clinical 1.5-T MR imaging technology. (orig.)

  18. Velocimetry using scintillation of a laser beam for a laser-based gas-flux monitor

    Kagawa, Naoki; Wada, Osami; Koga, Ryuji

    1999-05-01

    This paper describes a velocimetry system using scintillation of a laser-beam with spatial filters based on sensor arrays for a laser- based gas flux monitor. In the eddy correlation method, gas flux is obtained by mutual relation between the gas density and the flow velocity. The velocimetry system is developed to support the flow velocity monitor portion of the laser-based gas flux monitor with a long span for measurement. In order to sense not only the flow velocity but also the flow direction, two photo diode arrays are arranged with difference of a quarter period of the weighting function between them; the two output signals from the sensor arrays have phase difference of either (pi) /2 or -(pi) /2 depending on the sense of flow direction. In order to obtain the flow velocity and the flow direction instantly, an electronic apparatus built by the authors extracts frequency and phase from crude outputs of the pair of sensors. A feasibility of the velocimetry was confirmed indoors by measurement of the flow- velocity vector of the convection. Measured flow-velocity vector of the upward flow agreed comparatively with results of an ultrasonic anemometer.

  19. In-Vivo Detection and Tracking of T Cells in Various Organs in a Melanoma Tumor Model by 19F-Fluorine MRS/MRI.

    Christine Gonzales

    Full Text Available 19F-MRI and 19F-MRS can identify specific cell types after in-vitro or in-vivo 19F-labeling. Knowledge on the potential to track in-vitro 19F-labeled immune cells in tumor models by 19F-MRI/MRS is scarce.To study 19F-based MR techniques for in-vivo tracking of adoptively transferred immune cells after in-vitro 19F-labeling, i.e. to detect and monitor their migration non-invasively in melanoma-bearing mice.Splenocytes (SP were labeled in-vitro with a perfluorocarbon (PFC and IV-injected into non-tumor bearing mice. In-vitro PFC-labeled ovalbumin (OVA-specific T cells from the T cell receptor-transgenic line OT-1, activated with anti-CD3 and anti-CD28 antibodies (Tact or OVA-peptide pulsed antigen presenting cells (TOVA-act, were injected into B16 OVA melanoma-bearing mice. The distribution of the 19F-labelled donor cells was determined in-vivo by 19F-MRI/MRS. In-vivo 19F-MRI/MRS results were confirmed by ex-vivo 19F-NMR and flow cytometry.SP, Tact, and TOVA-act were successfully PFC-labeled in-vitro yielding 3x1011-1.4x1012 19F-atoms/cell in the 3 groups. Adoptively transferred 19F-labeled SP, TOVA-act, and Tact were detected by coil-localized 19F-MRS in the chest, abdomen, and left flank in most animals (corresponding to lungs, livers, and spleens, respectively, with highest signal-to-noise for SP vs TOVA-act and Tact, p<0.009 for both. SP and Tact were successfully imaged by 19F-MRI (n = 3; liver. These in-vivo data were confirmed by ex-vivo high-resolution 19F-NMR-spectroscopy. By flow cytometric analysis, however, TOVA-act tended to be more abundant versus SP and Tact (liver: p = 0.1313; lungs: p = 0.1073; spleen: p = 0.109. Unlike 19F-MRI/MRS, flow cytometry also identified transferred immune cells (SP, Tact, and TOVA-act in the tumors.SP, Tact, and TOVA-act were successfully PFC-labeled in-vitro and detected in-vivo by non-invasive 19F-MRS/MRI in liver, lung, and spleen. The portion of 19F-labeled T cells in the adoptively transferred cell

  20. Magentic Cell labeling of primary and stem cell-derived pig hepatocytes for MRI-based cell tracking of heptocytes transplantation

    Pig hepatocytes are an important investigational tool for optimizing hepatocyte transplantation schemes in both allogeneic and xenogeneic transplant scenarios. MRI can be used to serially monitor the transplanted cells, but only if the hepatocytes can be labeled with a magnetic particle. In this wo...

  1. Online Tracking

    ... can disable blocking on those sites. Tagged with: computer security , cookies , Do Not Track , personal information , privacy June ... email Looking for business guidance on privacy and ... The Federal Trade Commission (FTC) is the nation’s consumer protection agency. The FTC works to prevent fraudulent, deceptive ...

  2. Design, Construction and Effectiveness Analysis of Hybrid Automatic Solar Tracking System for Amorphous and Crystalline Solar Cells

    Bhupendra Gupta

    2013-01-01

    - This paper concerns the design and construction of a Hybrid solar tracking system. The constructed device was implemented by integrating it with Amorphous & Crystalline Solar Panel, three dimensional freedom mechanism and microcontroller. The amount of power available from a photovoltaic panel is determined by three parameters, the type of solar tracker, materials of solar panel and the intensity of the sunlight. The objective of this paper is to present analysis on the use of two differ...

  3. In vivo tracking of genetically engineered, anti-HER2/neu directed natural killer cells to HER2/neu positive mammary tumors with magnetic resonance imaging

    Daldrup-Link, Heike E.; Meier, Reinhardt; Metz, Stephan; Settles, Marcus; Rummeny, Ernst J.; Rudelius, Martina; Piontek, Guido; Schlegel, Juergen; Piert, Morand; Uherek, Christoph; Wels, Winfried

    2005-01-01

    agents, and the accumulation of these labeled cells in murine tumors can be monitored in vivo with MR imaging. This MR cell tracking technique may be applied to monitor NK-cell based immunotherapies in patients in order to assess the presence and extent of NK-cell tumor accumulations and, thus, to determine therapy response early and non-invasively. (orig.)

  4. In vivo tracking of genetically engineered, anti-HER2/neu directed natural killer cells to HER2/neu positive mammary tumors with magnetic resonance imaging

    Daldrup-Link, Heike E. [UCSF Medical Center, Department of Radiology, San Francisco, CA (United States); Meier, Reinhardt; Metz, Stephan; Settles, Marcus; Rummeny, Ernst J. [Technical University Munich, Department of Radiology, Munich (Germany); Rudelius, Martina; Piontek, Guido; Schlegel, Juergen [Technical University Munich, Institute of Pathology, Division of Neuropathology, Munich (Germany); Piert, Morand [Technical University Munich, Department of Nuclear Medicine, Munich (Germany); Uherek, Christoph; Wels, Winfried [University of Frankfurt, Georg Speyer House, Frankfurt (Germany)

    2005-01-01

    labeled with clinically applicable iron-oxide contrast agents, and the accumulation of these labeled cells in murine tumors can be monitored in vivo with MR imaging. This MR cell tracking technique may be applied to monitor NK-cell based immunotherapies in patients in order to assess the presence and extent of NK-cell tumor accumulations and, thus, to determine therapy response early and non-invasively. (orig.)

  5. NucliTrack: an integrated nuclei tracking application.

    Cooper, Sam; Barr, Alexis R; Glen, Robert; Bakal, Chris

    2017-10-15

    Live imaging studies give unparalleled insight into dynamic single cell behaviours and fate decisions. However, the challenge of reliably tracking single cells over long periods of time limits both the throughput and ease with which such studies can be performed. Here, we present NucliTrack, a cross platform solution for automatically segmenting, tracking and extracting features from fluorescently labelled nuclei. NucliTrack performs similarly to other state-of-the-art cell tracking algorithms, but NucliTrack's interactive, graphical interface makes it significantly more user friendly. NucliTrack is available as a free, cross platform application and open source Python package. Installation details and documentation are at: http://nuclitrack.readthedocs.io/en/latest/ A video guide can be viewed online: https://www.youtube.com/watch?v=J6e0D9F-qSU Source code is available through Github: https://github.com/samocooper/nuclitrack. A Matlab toolbox is also available at: https://uk.mathworks.com/matlabcentral/fileexchange/61479-samocooper-nuclitrack-matlab. sam@socooper.com. Supplementary data are available at Bioinformatics online. © The Author(s) 2017. Published by Oxford University Press.

  6. Measurement of 3-D Flow inside a Micro Curved-tube using Digital Micro Holographic Particle Tracking Velocimetry

    Lee, Sang Woo; Chae, Byoung Joo

    2007-01-01

    The effect of incidence angle on the three-dimensional flow and aerodynamic loss in the downstream region of a high-turning turbine rotor blade has been investigated with a straight miniature five-hole probe. The incidence angle is changed to be +10, +5, 0, -10, -20, -30 and -40 degrees. The results show that the positive incidence reinforces the three-dimensional vortical flows within the turbine passage including the passage vortex, but the negative incidence weaken them significantly. A small increment in the positive incidence angle results in a remarkable aerodynamic loss increase, while increasing the incidence angle in the negative range leads to a very small change in the aerodynamic loss

  7. A 2D/3D image analysis system to track fluorescently labeled structures in rod-shaped cells: application to measure spindle pole asymmetry during mitosis.

    Schmitter, Daniel; Wachowicz, Paulina; Sage, Daniel; Chasapi, Anastasia; Xenarios, Ioannis; Simanis; Unser, Michael

    2013-01-01

    The yeast Schizosaccharomyces pombe is frequently used as a model for studying the cell cycle. The cells are rod-shaped and divide by medial fission. The process of cell division, or cytokinesis, is controlled by a network of signaling proteins called the Septation Initiation Network (SIN); SIN proteins associate with the SPBs during nuclear division (mitosis). Some SIN proteins associate with both SPBs early in mitosis, and then display strongly asymmetric signal intensity at the SPBs in late mitosis, just before cytokinesis. This asymmetry is thought to be important for correct regulation of SIN signaling, and coordination of cytokinesis and mitosis. In order to study the dynamics of organelles or large protein complexes such as the spindle pole body (SPB), which have been labeled with a fluorescent protein tag in living cells, a number of the image analysis problems must be solved; the cell outline must be detected automatically, and the position and signal intensity associated with the structures of interest within the cell must be determined. We present a new 2D and 3D image analysis system that permits versatile and robust analysis of motile, fluorescently labeled structures in rod-shaped cells. We have designed an image analysis system that we have implemented as a user-friendly software package allowing the fast and robust image-analysis of large numbers of rod-shaped cells. We have developed new robust algorithms, which we combined with existing methodologies to facilitate fast and accurate analysis. Our software permits the detection and segmentation of rod-shaped cells in either static or dynamic (i.e. time lapse) multi-channel images. It enables tracking of two structures (for example SPBs) in two different image channels. For 2D or 3D static images, the locations of the structures are identified, and then intensity values are extracted together with several quantitative parameters, such as length, width, cell orientation, background fluorescence and

  8. Improvement of image velocimetry based on a spatio-temporal correlation method; Jikukan sokan ni motozuku ryushi gazo sokudoba keisokuho no kaiseki seino kaizen

    Yamada, H. [Tokuyama College of Technology, Yamaguchi (Japan); Arifuku, T. [Komatsu Ltd., Tokyo (Japan); Koga, K. [Yamaguchi University, Yamaguchi (Japan). Faculty of Engineering

    1998-05-31

    In the image velocimetry, it is generally required to detect the various velocity in each position of the flow field. But the maximum velocity which the usual velocimetry can detect has been limited in about 1 pixel per frame. Then, in order to measure the wide range of velocity vectors from the dynamic image, the improvement of performance in the image velocimetry based on a spatio-temporal correlation method would be attempted by enlarging the analytical region and by interpolating the new frame. The analytical performance of velocimetry was estimated by measuring the velocity from the flow dynamic image made artificially on the personal computer so as to simulate the flow of fluid containing a lot of small particles. As the results, the velocity range of the improved velocimetry became larger than that of the usual velocimetry. 21 refs., 13 figs., 1 tab.

  9. A REVIEW OF EXPERIMENTAL STUDY OF POWER QUALITY MEASUREMENT OF PHOTOVOLTAIC CELLS WITH MAXIMUM POWER POINT TRACKING SYSTEM

    Smriti Dwivedi; Prof. Sunil Kumar Bhatt

    2016-01-01

    Maximum power point tracking (MPPT) is a technique that charge controllers use for wind turbines and PV solar systems to maximize power output. PV solar systems exist in several different configurations. The most basic version sends power from collector panels directly to the DC-AC inverter and from there directly to the electrical grid. A second version, called a hybrid inverter, might split the power at the inverter, where a percentage of the power goes to the grid and the remainder goes to...

  10. Real-Time Tracking of BODIPY-C12 Long-Chain Fatty Acid in Human Term Placenta Reveals Unique Lipid Dynamics in Cytotrophoblast Cells.

    Kevin Kolahi

    Full Text Available While the human placenta must provide selected long-chain fatty acids to support the developing fetal brain, little is known about the mechanisms underlying the transport process. We tracked the movement of the fluorescently labeled long-chain fatty acid analogue, BODIPY-C12, across the cell layers of living explants of human term placenta. Although all layers took up the fatty acid, rapid esterification of long-chain fatty acids and incorporation into lipid droplets was exclusive to the inner layer cytotrophoblast cells rather than the expected outer syncytiotrophoblast layer. Cytotrophoblast is a progenitor cell layer previously relegated to a repair role. As isolated cytotrophoblasts differentiated into syncytialized cells in culture, they weakened their lipid processing capacity. Syncytializing cells suppress previously active genes that regulate fatty-acid uptake (SLC27A2/FATP2, FABP4, ACSL5 and lipid metabolism (GPAT3, LPCAT3. We speculate that cytotrophoblast performs a previously unrecognized role in regulating placental fatty acid uptake and metabolism.

  11. Intracellular dynamics and fate of polystyrene nanoparticles in A549 Lung epithelial cells monitored by image (cross-) correlation spectroscopy and single particle tracking.

    Deville, Sarah; Penjweini, Rozhin; Smisdom, Nick; Notelaers, Kristof; Nelissen, Inge; Hooyberghs, Jef; Ameloot, Marcel

    2015-10-01

    Novel insights in nanoparticle (NP) uptake routes of cells, their intracellular trafficking and subcellular targeting can be obtained through the investigation of their temporal and spatial behavior. In this work, we present the application of image (cross-) correlation spectroscopy (IC(C)S) and single particle tracking (SPT) to monitor the intracellular dynamics of polystyrene (PS) NPs in the human lung carcinoma A549 cell line. The ensemble kinetic behavior of NPs inside the cell was characterized by temporal and spatiotemporal image correlation spectroscopy (TICS and STICS). Moreover, a more direct interpretation of the diffusion and flow detected in the NP motion was obtained by SPT by monitoring individual NPs. Both techniques demonstrate that the PS NP transport in A549 cells is mainly dependent on microtubule-assisted transport. By applying spatiotemporal image cross-correlation spectroscopy (STICCS), the correlated motions of NPs with the early endosomes, late endosomes and lysosomes are identified. PS NPs were equally distributed among the endolysosomal compartment during the time interval of the experiments. The cotransport of the NPs with the lysosomes is significantly larger compared to the other cell organelles. In the present study we show that the complementarity of ICS-based techniques and SPT enables a consistent elaborate model of the complex behavior of NPs inside biological systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Advances in using MRI probes and sensors for in vivo cell tracking as applied to regenerative medicine.

    Srivastava, Amit K; Kadayakkara, Deepak K; Bar-Shir, Amnon; Gilad, Assaf A; McMahon, Michael T; Bulte, Jeff W M

    2015-04-01

    The field of molecular and cellular imaging allows molecules and cells to be visualized in vivo non-invasively. It has uses not only as a research tool but in clinical settings as well, for example in monitoring cell-based regenerative therapies, in which cells are transplanted to replace degenerating or damaged tissues, or to restore a physiological function. The success of such cell-based therapies depends on several critical issues, including the route and accuracy of cell transplantation, the fate of cells after transplantation, and the interaction of engrafted cells with the host microenvironment. To assess these issues, it is necessary to monitor transplanted cells non-invasively in real-time. Magnetic resonance imaging (MRI) is a tool uniquely suited to this task, given its ability to image deep inside tissue with high temporal resolution and sensitivity. Extraordinary efforts have recently been made to improve cellular MRI as applied to regenerative medicine, by developing more advanced contrast agents for use as probes and sensors. These advances enable the non-invasive monitoring of cell fate and, more recently, that of the different cellular functions of living cells, such as their enzymatic activity and gene expression, as well as their time point of cell death. We present here a review of recent advancements in the development of these probes and sensors, and of their functioning, applications and limitations. © 2015. Published by The Company of Biologists Ltd.

  13. Superparamagnetic iron oxide nanoparticles function as a long-term, multi-modal imaging label for non-invasive tracking of implanted progenitor cells.

    Christina A Pacak

    Full Text Available The purpose of this study was to determine the ability of superparamagnetic iron oxide (SPIO nanoparticles to function as a long-term tracking label for multi-modal imaging of implanted engineered tissues containing muscle-derived progenitor cells using magnetic resonance imaging (MRI and X-ray micro-computed tomography (μCT. SPIO-labeled primary myoblasts were embedded in fibrin sealant and imaged to obtain intensity data by MRI or radio-opacity information by μCT. Each imaging modality displayed a detection gradient that matched increasing SPIO concentrations. Labeled cells were then incorporated in fibrin sealant, injected into the atrioventricular groove of rat hearts, and imaged in vivo and ex vivo for up to 1 year. Transplanted cells were identified in intact animals and isolated hearts using both imaging modalities. MRI was better able to detect minuscule amounts of SPIO nanoparticles, while μCT more precisely identified the location of heavily-labeled cells. Histological analyses confirmed that iron oxide particles were confined to viable, skeletal muscle-derived cells in the implant at the expected location based on MRI and μCT. These analyses showed no evidence of phagocytosis of labeled cells by macrophages or release of nanoparticles from transplanted cells. In conclusion, we established that SPIO nanoparticles function as a sensitive and specific long-term label for MRI and μCT, respectively. Our findings will enable investigators interested in regenerative therapies to non-invasively and serially acquire complementary, high-resolution images of transplanted cells for one year using a single label.

  14. Tracking of In-111-labeled human umbilical tissue-derived cells (hUTC) in a rat model of cerebral ischemia using SPECT imaging

    Arbab, Ali S; Chopp, Michael; Thiffault, Christine; Navia, Bradford; Victor, Stephen J; Hong, Klaudyne; Zhang, Li; Jiang, Quan; Varma, Nadimpalli RS; Iskander, ASM

    2012-01-01

    In order to increase understanding of how infused cells work, it becomes important to track their initial movement, localization, and engraftment efficiency following transplantation. However, the available in vivo cell tracking techniques are suboptimal. The study objective was to determine the biodistribution of intravenously administered Indium-111 (In-111) oxine labeled human umbilical tissue-derived cells (hUTC) in a rat model of transient middle cerebral occlusion (tMCAo) using single photon emission computed tomography (SPECT). Rats received 3 million In-111 labeled hUTC (i.v.) 48 hrs after tMCAo. Following the administration of either hUTC or equivalent dose of In-111-oxine (18.5 MBq), animals underwent SPECT imaging on days 0, 1, and 3. Radioactivity in various organs as well as in the stroke area and contralateral hemisphere was determined, decay corrected and normalized to the total (whole body including head) radioactivity on day 0. Immunohistochemical analysis was also performed to confirm the beneficial effects of hUTC on vascular and synaptic density, and apoptosis. Most of the radioactivity (43.36±23.07% on day 0) trafficked to the lungs immediately following IV administration of In-111 labeled hUTC (day 0) and decreased drastically to 8.81±7.75 and 4.01±4.52% on days 1 and 3 post-injection, respectively. In contrast, radioactivity measured in the lung of animals that received In-111-oxine alone remained relatively unchanged from day 0 to day 1 (18.38±5.45% at day 0 to 12.59±5.94%) and decreased to 8.34±4.25% on day 3. Significantly higher radioactivity was observed in stroke areas of animals that received In-111 labeled hUTC indicating the presence of cells at the site of injury representing approximately 1% of total administered dose. In addition, there was significant increase in vascular and synaptophysin immunoreactivity in stroke areas of rats that received In-111 labeled hUTC. The present studies showed the tracking of In-111 labeled h

  15. Evaluation of gold nanotracers to track adipose-derived stem cells in a PEGylated fibrin gel for dermal tissue engineering applications

    Chung E

    2013-01-01

    Full Text Available Eunna Chung,1 Seung Yun Nam,1,2 Laura M Ricles,1 Stanislav Y Emelianov,1,2 Laura J Suggs11Department of Biomedical Engineering, 2Department of Electrical and Computer Engineering, The University of Texas at Austin, Austin, TX, USAAbstract: Evaluating the regenerative capacity of a tissue-engineered device in a noninvasive and synchronous manner is critical to determining the mechanisms for success in clinical applications. In particular, directly tracking implanted cells in a three-dimensional (3D scaffold is desirable in that it enables the monitoring of cellular activity in a specific and localized manner. The authors' group has previously demonstrated that the PEGylation of fibrin results in a 3D scaffold that supports morphologic and phenotypic changes in mesenchymal stem cells that may be advantageous in wound healing applications. Recently, the authors have evaluated adipose-derived stem cells (ASCs as a mesenchymal cell source to regenerate skin and blood vessels due to their potential for proliferation, differentiation, and production of growth factors. However, tracking and monitoring ASCs in a 3D scaffold, such as a PEGylated fibrin gel, have not yet been fully investigated. In the current paper, nanoscale gold spheres (20 nm as cell tracers for ASCs cultured in a PEGylated fibrin gel were evaluated. An advanced dual-imaging modality combining ultrasound and photoacoustic imaging was utilized to monitor rat ASCs over time. The ASCs took up gold nanotracers and could be detected up to day 16 with high sensitivity using photoacoustic imaging. There were no detrimental effects on ASC morphology, network formation, proliferation, and protein expression/secretion (ie, smooth muscle α-actin, vascular endothelial growth factor, matrix metalloproteinase-2, and matrix metalloproteinase-9 associated with gold nanotracers. Therefore, utilization of gold nanotracers can be an effective strategy to monitor the regenerative process of a stem cell

  16. Steep Dose-Response Relationship for Stage I Non-Small-Cell Lung Cancer Using Hypofractionated High-Dose Irradiation by Real-Time Tumor-Tracking Radiotherapy

    Onimaru, Rikiya; Fujino, Masaharu; Yamazaki, Koichi; Onodera, Yuya; Taguchi, Hiroshi; Katoh, Norio; Hommura, Fumihiro; Oizumi, Satoshi; Nishimura, Masaharu; Shirato, Hiroki

    2008-01-01

    Purpose: To investigate the clinical outcomes of patients with pathologically proven, peripherally located, Stage I non-small-cell lung cancer who had undergone stereotactic body radiotherapy using real-time tumor tracking radiotherapy during the developmental period. Methods and Materials: A total of 41 patients (25 with Stage T1 and 16 with Stage T2) were admitted to the study between February 2000 and June 2005. A 5-mm planning target volume margin was added to the clinical target volume determined with computed tomography at the end of the expiratory phase. The gating window ranged from ±2 to 3 mm. The dose fractionation schedule was 40 or 48 Gy in four fractions within 1 week. The dose was prescribed at the center of the planning target volume, giving more than an 80% dose at the planning target volume periphery. Results: For 28 patients treated with 48 Gy in four fractions, the overall actuarial survival rate at 3 years was 82% for those with Stage IA and 32% for those with Stage IB. For patients treated with 40 Gy in four fractions within 1 week, the overall actuarial survival rate at 3 years was 50% for those with Stage IA and 0% for those with Stage IB. A significant difference was found in local control between those with Stage IB who received 40 Gy vs. 48 Gy (p = 0.0015) but not in those with Stage IA (p = 0.5811). No serious radiation morbidity was observed with either dose schedule. Conclusion: The results of our study have shown that 48 Gy in four fractions within 1 week is a safe and effective treatment for peripherally located, Stage IA non-small-cell lung cancer. A steep dose-response curve between 40 and 48 Gy using a daily dose of 12 Gy delivered within 1 week was identified for Stage IB non-small-cell lung cancer in stereotactic body radiotherapy using real-time tumor tracking radiotherapy

  17. Use of track-end alpha particles from 241Am to study radiosensitive sites in CHO cells

    Datta, R.; Cole, A.; Robinson, S.

    1976-01-01

    Monolayers of CHO cells placed on membrane filters were irradiated with alpha particles from a 241 Am source. Particle penetration into the cells was controlled by placing the cell sample at various distances from the source. Dosimetric and spectrometric measurements were performed at comparable positions using a parallel plate ionization chamber and a scintillation crystal spectrometer. Cell survival, as measured by conventional cloning techniques, was single hit in form. A pronounced minimum in mean lethal dose of 29 rad was observed for alpha particle beams that penetrated only about 3 μm into the cell. A pronounced maximum in inactivation cross section of 90 μm 2 , equal to about half the projected area of the nucleus, occurred for beams that penetrated only 5 to 7 μm into the cell. Thus, a single alpha particle penetration several micrometers within the cell nucleus was effective in killing the cell, while fully penetrating beams were actually less efficient; the latter beams required multiple particle traversals and about three times the cell dose to achieve the same effect. These results support the proposal that radiosensitive sites are located in a thin peripheral region of the nucleus

  18. ultraLM and miniLM: Locator tools for smart tracking of fluorescent cells in correlative light and electron microscopy.

    Brama, Elisabeth; Peddie, Christopher J; Wilkes, Gary; Gu, Yan; Collinson, Lucy M; Jones, Martin L

    2016-12-13

    In-resin fluorescence (IRF) protocols preserve fluorescent proteins in resin-embedded cells and tissues for correlative light and electron microscopy, aiding interpretation of macromolecular function within the complex cellular landscape. Dual-contrast IRF samples can be imaged in separate fluorescence and electron microscopes, or in dual-modality integrated microscopes for high resolution correlation of fluorophore to organelle. IRF samples also offer a unique opportunity to automate correlative imaging workflows. Here we present two new locator tools for finding and following fluorescent cells in IRF blocks, enabling future automation of correlative imaging. The ultraLM is a fluorescence microscope that integrates with an ultramicrotome, which enables 'smart collection' of ultrathin sections containing fluorescent cells or tissues for subsequent transmission electron microscopy or array tomography. The miniLM is a fluorescence microscope that integrates with serial block face scanning electron microscopes, which enables 'smart tracking' of fluorescent structures during automated serial electron image acquisition from large cell and tissue volumes.

  19. Tracking and Functional Characterization of Epithelial-Mesenchymal Transition and Mesenchymal Tumor Cells During Prostate Cancer Metastasis

    Ruscetti, Marcus; Quach, Bill; Dadashian, Eman L.; Mulholland, David J.; Wu, Hong

    2015-01-01

    The epithelial-mesenchymal transition (EMT) has been postulated as a mechanism by which cancer cells acquire the invasive and stem-like traits necessary for distant metastasis. However, direct in vivo evidence for the role of EMT in the formation of cancer stem-like cells (CSC) and the metastatic cascade remains lacking. Here we report the first isolation and characterization of mesenchymal and EMT tumor cells, which harbor both epithelial and mesenchymal characteristics, in an autochthonous murine model of prostate cancer. By crossing the established Pb-Cre+/−;PtenL/L;KrasG12D/+ prostate cancer model with a vimentin-GFP reporter strain, generating CPKV mice, we were able to isolate epithelial, EMT and mesenchymal cancer cells based on expression of vimentin and EpCAM. CPKV mice (but not mice with Pten deletion alone) exhibited expansion of cells with EMT (EpCAM+/Vim-GFP+) and mesenchymal (EpCAM−/Vim-GFP+) characteristics at the primary tumor site and in circulation. These EMT and mesenchymal tumor cells displayed enhanced stemness and invasive character compared to epithelial tumor cells. Moreover, they displayed an enriched tumor-initiating capacity and could regenerate epithelial glandular structures in vivo, indicative of epithelia-mesenchyme plasticity. Interestingly, while mesenchymal tumor cells could persist in circulation and survive in the lung following intravenous injection, only epithelial and EMT tumor cells could form macrometastases. Our work extends the evidence that mesenchymal and epithelial states in cancer cells contribute differentially to their capacities for tumor initiation and metastatic seeding, respectively, and that EMT tumor cells exist with plasticity that can contribute to multiple stages of the metastatic cascade. PMID:25948589

  20. Flow-Tagging Velocimetry for Hypersonic Flows Using Fluorescence of Nitric Oxide

    Danehy, P. M.; OByrne, S.; Houwing, A. F. P.

    2001-01-01

    We investigate a new type of flow-tagging velocimetry technique for hypersonic flows. The technique involves exciting a thin line of nitric oxide molecules with a laser beam and then, after some delay, acquiring an image of the displaced line. One component of velocity is determined from the time of flight. This method is applied to measure the velocity profile in a Mach 8.5 laminar, hypersonic boundary layer in the Australian National Universities T2 free-piston shock tunnel. The velocity is measured with an uncertainty of approximately 2%. Comparison with a CFD simulation of the flow shows reasonable agreement.

  1. Spatial filtering velocimetry of objective speckles for measuring out-of-plane motion

    Jakobsen, Michael Linde; Yura, H. T.; Hanson, Steen Grüner

    2012-01-01

    This paper analyzes the dynamics of objective laser speckles as the distance between the object and the observation plane continuously changes. With the purpose of applying optical spatial filtering velocimetry to the speckle dynamics, in order to measure out-of-plane motion in real time......, a rotational symmetric spatial filter is designed. The spatial filter converts the speckle dynamics into a photocurrent with a quasi-sinusoidal response to the out-of-plane motion. The spatial filter is here emulated with a CCD camera, and is tested on speckles arising from a real application. The analysis...

  2. Surge Flow in a Centrifugal Compressor Measured by Digital Particle Image Velocimetry

    Wernet, Mark P.

    2000-01-01

    A planar optical velocity measurement technique known as Particle Image Velocimetry (PIV) is being used to study transient events in compressors. In PIV, a pulsed laser light sheet is used to record the positions of particles entrained in a fluid at two instances in time across a planar region of the flow. Determining the recorded particle displacement between exposures yields an instantaneous velocity vector map across the illuminated plane. Detailed flow mappings obtained using PIV in high-speed rotating turbomachinery components are used to improve the accuracy of computational fluid dynamics (CFD) simulations, which in turn, are used to guide advances in state-of-the-art aircraft engine hardware designs.

  3. New Developments In Particle Image Velocimetry (PIV) For The Study Of Complex Plasmas

    Thomas, Edward Jr.; Fisher, Ross; Shaw, Joseph; Jefferson, Robert; Cianciosa, Mark; Williams, Jeremiah

    2011-01-01

    Particle Image Velocimetry (PIV) is a fluid measurement technique in which the average displacement of small groups of particles is made by comparing a pair of images that are separated in time by an interval Δt. For over a decade, a several variations of the PIV technique, e.g., two-dimensional, stereoscopic, and tomographic PIV, have been used to characterize particle transport, instabilities, and the thermal properties of complex plasmas. This paper describes the basic principles involved in the PIV analysis technique and discusses potential future applications of PIV to the study of complex plasmas.

  4. Correcting for particle size effects on plasma actuator particle image velocimetry measurements

    Masati, A.; Sedwick, R. J.

    2018-01-01

    Particle image velocimetry (PIV) is often used to characterize plasma actuator flow, but particle charging effects are rarely taken into account. A parametric study was conducted to determine the effects of particle size on the velocity results of plasma actuator PIV experiments. Results showed that smaller particles more closely match air flow velocities than larger particles. The measurement uncertainty was quantified by deconvolving the particle image diameter from the correlation diameter. The true air velocity was calculated by linearly extrapolating to the zero-size particle diameter.

  5. Time-resolved Particle Image Velocimetry measurements of the 3D random Richtmyer-Meshkov Instability

    Sewell, Everest; Krivets, Vitaliy; Jacobs, Jeffrey

    2017-11-01

    The vertical shock tube at the University of Arizona is used to perform experiments on the multi-mode three-dimensional Richtmyer-Meshkov Instability (RMI). An interface of air and sulfur hexafluoride is formed in a counter flow configuration, and is excited using voice coils to produce faraday-like multi-modal perturbations.This interface is shock accelerated by an approximately Mach 1.2 shockwave to form the RMI. Time resolved Particle Image Velocimetry (PIV) is used to perform analysis of the evolving instability.

  6. GTSE1 is a microtubule plus-end tracking protein that regulates EB1-dependent cell migration.

    Massimilano Scolz

    Full Text Available The regulation of cell migration is a highly complex process that is often compromised when cancer cells become metastatic. The microtubule cytoskeleton is necessary for cell migration, but how microtubules and microtubule-associated proteins regulate multiple pathways promoting cell migration remains unclear. Microtubule plus-end binding proteins (+TIPs are emerging as important players in many cellular functions, including cell migration. Here we identify a +TIP, GTSE1, that promotes cell migration. GTSE1 accumulates at growing microtubule plus ends through interaction with the EB1+TIP. The EB1-dependent +TIP activity of GTSE1 is required for cell migration, as well as for microtubule-dependent disassembly of focal adhesions. GTSE1 protein levels determine the migratory capacity of both nontransformed and breast cancer cell lines. In breast cancers, increased GTSE1 expression correlates with invasive potential, tumor stage, and time to distant metastasis, suggesting that misregulation of GTSE1 expression could be associated with increased invasive potential.

  7. Characterization of the early events in dengue virus cell entry by biochemical assays and single-virus tracking

    van der Schaar, Hilde M.; Rust, Michael J.; Waarts, Barry-Lee; van der Ende-Metselaarl, Heidi; Kuhn, Richard J.; Wilschut, Jan; Zhuang, Xiaowei; Smit, Jolanda M.

    In this study, we investigated the cell entry characteristics of dengue virus (DENV) type 2 strain SI on mosquito, BHK-15, and BS-C-1 cells. The concentration of virus particles measured by biochemical assays was found to be substantially higher than the number of infectious particles determined by

  8. Characterization of the early events in dengue virus cell entry by biochemical assays and single-virus tracking

    van der Schaar, Hilde M.; Rust, Michael J.; Waarts, Barry-Lee; van der Ende-Metselaarl, Heidi; Kuhn, Richard J.; Wilschut, Jan; Zhuang, Xiaowei; Smit, Jolanda M.

    2007-01-01

    In this study, we investigated the cell entry characteristics of dengue virus (DENV) type 2 strain SI on mosquito, BHK-15, and BS-C-1 cells. The concentration of virus particles measured by biochemical assays was found to be substantially higher than the number of infectious particles determined by

  9. Motion tracking to enable pre-surgical margin mapping in basal cell carcinoma using optical imaging modalities: initial feasibility study using optical coherence tomography

    Duffy, M.; Richardson, T. J.; Craythorne, E.; Mallipeddi, R.; Coleman, A. J.

    2014-02-01

    A system has been developed to assess the feasibility of using motion tracking to enable pre-surgical margin mapping of basal cell carcinoma (BCC) in the clinic using optical coherence tomography (OCT). This system consists of a commercial OCT imaging system (the VivoSight 1500, MDL Ltd., Orpington, UK), which has been adapted to incorporate a webcam and a single-sensor electromagnetic positional tracking module (the Flock of Birds, Ascension Technology Corp, Vermont, USA). A supporting software interface has also been developed which allows positional data to be captured and projected onto a 2D dermoscopic image in real-time. Initial results using a stationary test phantom are encouraging, with maximum errors in the projected map in the order of 1-2mm. Initial clinical results were poor due to motion artefact, despite attempts to stabilise the patient. However, the authors present several suggested modifications that are expected to reduce the effects of motion artefact and improve the overall accuracy and clinical usability of the system.

  10. Integrating UF6 Cylinder RF Tracking With Continuous Load Cell Monitoring for Verifying Declared UF6 Feed and Withdrawal Operations Verifying Declared UF6 Feed and Withdrawal Operations

    Krichinsky, Alan M.; Miller, Paul; Pickett, Chris A.; Richardson, Dave; Rowe, Nathan C.; Whitaker, J. Michael; Younkin, James R.

    2009-01-01

    Oak Ridge National Laboratory is demonstrating the integration of UF6 cylinder tracking, using RF technology, with continuous load cell monitoring (CLCM) at mock UF6 feed and withdrawal (F and W) stations. CLCM and cylinder tracking are two of several continuous-monitoring technologies that show promise in providing integrated safeguards of F and W operations at enrichment plants. Integrating different monitoring technologies allows advanced, automated event processing to screen innocuous events thereby minimizing false alerts to independent inspectors. Traditionally, international inspectors rely on batch verification of material inputs and outputs derived from operator declarations and periodic on-site inspections at uranium enrichment plants or other nuclear processing facilities. Continuously monitoring F and W activities between inspections while providing filtered alerts of significant operational events will substantially increase the amount of valuable information available to inspectors thereby promising to enhance the effectiveness of safeguards and to improve efficiency in conducting on-site inspections especially at large plants for ensuring that all operations are declared.

  11. Latent tracks in polymeric etched track detectors

    Yamauchi, Tomoya

    2013-01-01

    Track registration properties in polymeric track detectors, including Poly(allyl diglycol carbonate), Bispenol A polycarbonate, Poly(ethylen terephtarate), and Polyimide, have been investigated by means of Fourie transform Infararede FT-IR spectrometry. Chemical criterion on the track formation threshold has been proposes, in stead of the conventional physical track registration models. (author)

  12. Tracking telecommuting

    Stastny, P.

    2007-03-15

    Many employees are now choosing to work from home using laptops and telephones. Employers in the oil and gas industry are now reaping a number of benefits from their telecommuting employees, including increased productivity; higher levels of employee satisfaction, and less absenteeism. Providing a telecommunication option can prove to be advantageous for employers wishing to hire or retain employees. Telecommuting may also help to reduce greenhouse gas (GHG) emissions. This article provided details of Teletrips Inc., a company that aids in the production of corporate social responsibility reports. Teletrips provides reports that document employee savings in time, vehicle depreciation maintenance, and gasoline costs. Teletrips currently tracks 12 companies in Calgary, and plans to grow through the development of key technology partnerships. The company is also working with the federal government to provide their clients with emission trading credits, and has forged a memorandum of understanding with the British Columbia government for tracking emissions. Calgary now openly supports telecommuting and is encouraging businesses in the city to adopt telecommuting on a larger scale. It was concluded that the expanding needs for road infrastructure and the energy used by cars to move workers in and out of the city are a massive burden to the city's tax base. 1 fig.

  13. INNER TRACKING

    P. Sharp

    The CMS Inner Tracking Detector continues to make good progress. The Objective for 2006 was to complete all of the CMS Tracker sub-detectors and to start the integration of the sub-detectors into the Tracker Support Tube (TST). The Objective for 2007 is to deliver to CMS a completed, installed, commissioned and calibrated Tracking System (Silicon Strip and Pixels) aligned to < 100µ in April 2008 ready for the first physics collisions at LHC. In November 2006 all of the sub-detectors had been delivered to the Tracker Integration facility (TIF) at CERN and the tests and QA procedures to be carried out on each sub-detector before integration had been established. In December 2006, TIB/TID+ was integrated into TOB+, TIB/TID- was being prepared for integration, and TEC+ was undergoing tests at the final tracker operating temperature (-100 C) in the Lyon cold room. In February 2007, TIB/TID- has been integrated into TOB-, and the installation of the pixel support tube and the services for TI...

  14. Application of magnetic carriers to two examples of quantitative cell analysis

    Zhou, Chen; Qian, Zhixi; Choi, Young Suk; David, Allan E. [Department of Chemical Engineering, 212 Ross Hall, Auburn University, Auburn, AL 36849 (United States); Todd, Paul, E-mail: pwtodd@hotmail.com [Techshot, Inc., 7200 Highway 150, Greenville, IN 47124 (United States); Hanley, Thomas R. [Department of Chemical Engineering, 212 Ross Hall, Auburn University, Auburn, AL 36849 (United States)

    2017-04-01

    The use of magnetophoretic mobility as a surrogate for fluorescence intensity in quantitative cell analysis was investigated. The objectives of quantitative fluorescence flow cytometry include establishing a level of labeling for the setting of parameters in fluorescence activated cell sorters (FACS) and the determination of levels of uptake of fluorescently labeled substrates by living cells. Likewise, the objectives of quantitative magnetic cytometry include establishing a level of labeling for the setting of parameters in flowing magnetic cell sorters and the determination of levels of uptake of magnetically labeled substrates by living cells. The magnetic counterpart to fluorescence intensity is magnetophoretic mobility, defined as the velocity imparted to a suspended cell per unit of magnetic ponderomotive force. A commercial velocimeter available for making this measurement was used to demonstrate both applications. Cultured Gallus lymphoma cells were immunolabeled with commercial magnetic beads and shown to have adequate magnetophoretic mobility to be separated by a novel flowing magnetic separator. Phagocytosis of starch nanoparticles having magnetic cores by cultured Chinese hamster ovary cells, a CHO line, was quantified on the basis of magnetophoretic mobility. - Highlights: • Commercial particle tracking velocimetry measures magnetophoretic mobility of labeled cells. • Magnetically labeled tumor cells were shown to have adequate mobility for capture in a specific sorter. • The kinetics of nonspecific endocytosis of magnetic nanomaterials by CHO cells was characterized. • Magnetic labeling of cells can be used like fluorescence flow cytometry for quantitative cell analysis.

  15. In-Vivo Imaging Of Transplanted Human Hepatic Stem Cells: Negative Contrast Labeling And 7t Micro-MRI Tracking

    McClelland, Randall E; Wauthier, Eliane; Reid, Lola; Hsu, Edward

    2004-01-01

    Stem cell therapies have great potential as alternative options to whole organ transplantations in treating dysfunction or failure, and alleviating the chronic shortage of donor availability of organs such as the...

  16. Tracking multi-walled carbon nanotubes inside oat (Avena sativa L.) plants and assessing their effect on growth, yield, and mammalian (human) cell viability

    Joshi, Anjali; Kaur, Simranjeet; Singh, Pargat; Dharamvir, Keya; Nayyar, Harsh; Verma, Gaurav

    2018-05-01

    Our findings show that oxidized multi-walled carbon nanotubes (MWCNT) having serpent-like morphology and smaller sizes (diameter of 35 nm and lengths of 200-300 nm) are compatible with oat plant tissues. Applied by seed-priming method as 90 µg/ml concentration, these serpentine MWCNT (having open-end caps) enter the oat plant and traverse the cells. Tracking of MWCNT inside sections and tissues during growth of oat plant has been done using special sample preparation. We present clear images of MWCNT inside the primed seeds and vascular bundles, the conducting tissues of root and shoot of oat. A dye fluorescein isothiocyanate non-covalently bonded to MWCNT also helped in detecting the path through circumferential perimeters of the oat channels, using fluorescence and confocal microscopy. The presence of MWCNT inside oat enhanced the growth of xylem cells by about 1.85-fold in vasculature of shoots. Compared to controls, the chlorophyll content increased by 57%, while photosynthetic activity enhanced by 15% for the same sample in MWCNT-primed plants. Overall, the growth factors were also augmented leading to significant increase in yield components. No toxic effects of MWCNT were observed in the DNA of the primed plants, and in the human cell lines treated with grains harvested from the MWCNT-primed plants. Our study provides some new insights about the role of MWCNT in plants and their potential benefits in agriculture.

  17. Longitudinal tracking of subpopulation dynamics and molecular changes during LNCaP cell castration and identification of inhibitors that could target the PSA−/lo castration-resistant cells

    Rycaj, Kiera; Cho, Eun Jeong; Liu, Xin; Chao, Hsueh-Ping; Liu, Bigang; Li, Qiuhui; Devkota, Ashwini K.; Zhang, Dingxiao; Chen, Xin; Moore, John; Dalby, Kevin N.; Tang, Dean G.

    2016-01-01

    We have recently demonstrated that the undifferentiated PSA−/lo prostate cancer (PCa) cell population harbors self-renewing long-term tumor-propagating cells that are refractory to castration, thus representing a therapeutic target. Our goals here are, by using the same lineage-tracing reporter system, to track the dynamic changes of PSA−/lo and PSA+ cells upon castration in vitro, investigate the molecular changes accompanying persistent castration, and develop large numbers of PSA−/lo PCa cells for drug screening. To these ends, we treated LNCaP cells infected with the PSAP-GFP reporter with three regimens of castration, i.e., CDSS, CDSS plus bicalutamide, and MDV3100 continuously for up to ~21 months. We observed that in the first ~7 months, castration led to time-dependent increases in PSA−/lo cells, loss of AR and PSA expression, increased expression of cancer stem cell markers, and many other molecular changes. Meanwhile, castrated LNCaP cells became resistant to high concentrations of MDV3100, chemotherapeutic drugs, and other agents. However, targeted and medium-throughput library screening identified several kinase (e.g., IGF-1R, AKT, PI3K/mTOR, Syk, GSK3) inhibitors as well as the BCL2 inhibitor that could effectively sensitize the LNCaP-CRPC cells to killing. Of interest, LNCaP cells castrated for >7 months showed evidence of cyclic changes in AR and the mTOR/AKT signaling pathways potentially involving epigenetic mechanisms. These observations indicate that castration elicits numerous molecular changes and leads to enrichment of PSA−/lo PCa cells. The ability to generate large numbers of PSA−/lo PCa cells should allow future high-throughput screening to identify novel therapeutics that specifically target this population. PMID:26871947

  18. Fundamental uncertainty limit of optical flow velocimetry according to Heisenberg's uncertainty principle.

    Fischer, Andreas

    2016-11-01

    Optical flow velocity measurements are important for understanding the complex behavior of flows. Although a huge variety of methods exist, they are either based on a Doppler or a time-of-flight measurement principle. Doppler velocimetry evaluates the velocity-dependent frequency shift of light scattered at a moving particle, whereas time-of-flight velocimetry evaluates the traveled distance of a scattering particle per time interval. Regarding the aim of achieving a minimal measurement uncertainty, it is unclear if one principle allows to achieve lower uncertainties or if both principles can achieve equal uncertainties. For this reason, the natural, fundamental uncertainty limit according to Heisenberg's uncertainty principle is derived for Doppler and time-of-flight measurement principles, respectively. The obtained limits of the velocity uncertainty are qualitatively identical showing, e.g., a direct proportionality for the absolute value of the velocity to the power of 32 and an indirect proportionality to the square root of the scattered light power. Hence, both measurement principles have identical potentials regarding the fundamental uncertainty limit due to the quantum mechanical behavior of photons. This fundamental limit can be attained (at least asymptotically) in reality either with Doppler or time-of-flight methods, because the respective Cramér-Rao bounds for dominating photon shot noise, which is modeled as white Poissonian noise, are identical with the conclusions from Heisenberg's uncertainty principle.

  19. Reconstruction of an acoustic pressure field in a resonance tube by particle image velocimetry.

    Kuzuu, K; Hasegawa, S

    2015-11-01

    A technique for estimating an acoustic field in a resonance tube is suggested. The estimation of an acoustic field in a resonance tube is important for the development of the thermoacoustic engine, and can be conducted employing two sensors to measure pressure. While this measurement technique is known as the two-sensor method, care needs to be taken with the location of pressure sensors when conducting pressure measurements. In the present study, particle image velocimetry (PIV) is employed instead of a pressure measurement by a sensor, and two-dimensional velocity vector images are extracted as sequential data from only a one- time recording made by a video camera of PIV. The spatial velocity amplitude is obtained from those images, and a pressure distribution is calculated from velocity amplitudes at two points by extending the equations derived for the two-sensor method. By means of this method, problems relating to the locations and calibrations of multiple pressure sensors are avoided. Furthermore, to verify the accuracy of the present method, the experiments are conducted employing the conventional two-sensor method and laser Doppler velocimetry (LDV). Then, results by the proposed method are compared with those obtained with the two-sensor method and LDV.

  20. High Dynamic Velocity Range Particle Image Velocimetry Using Multiple Pulse Separation Imaging

    Tadhg S. O’Donovan

    2010-12-01

    Full Text Available The dynamic velocity range of particle image velocimetry (PIV is determined by the maximum and minimum resolvable particle displacement. Various techniques have extended the dynamic range, however flows with a wide velocity range (e.g., impinging jets still challenge PIV algorithms. A new technique is presented to increase the dynamic velocity range by over an order of magnitude. The multiple pulse separation (MPS technique (i records series of double-frame exposures with different pulse separations, (ii processes the fields using conventional multi-grid algorithms, and (iii yields a composite velocity field with a locally optimized pulse separation. A robust criterion determines the local optimum pulse separation, accounting for correlation strength and measurement uncertainty. Validation experiments are performed in an impinging jet flow, using laser-Doppler velocimetry as reference measurement. The precision of mean flow and turbulence quantities is significantly improved compared to conventional PIV, due to the increase in dynamic range. In a wide range of applications, MPS PIV is a robust approach to increase the dynamic velocity range without restricting the vector evaluation methods.

  1. High dynamic velocity range particle image velocimetry using multiple pulse separation imaging.

    Persoons, Tim; O'Donovan, Tadhg S

    2011-01-01

    The dynamic velocity range of particle image velocimetry (PIV) is determined by the maximum and minimum resolvable particle displacement. Various techniques have extended the dynamic range, however flows with a wide velocity range (e.g., impinging jets) still challenge PIV algorithms. A new technique is presented to increase the dynamic velocity range by over an order of magnitude. The multiple pulse separation (MPS) technique (i) records series of double-frame exposures with different pulse separations, (ii) processes the fields using conventional multi-grid algorithms, and (iii) yields a composite velocity field with a locally optimized pulse separation. A robust criterion determines the local optimum pulse separation, accounting for correlation strength and measurement uncertainty. Validation experiments are performed in an impinging jet flow, using laser-Doppler velocimetry as reference measurement. The precision of mean flow and turbulence quantities is significantly improved compared to conventional PIV, due to the increase in dynamic range. In a wide range of applications, MPS PIV is a robust approach to increase the dynamic velocity range without restricting the vector evaluation methods.

  2. Tracking Porters

    Bruun, Maja Hojer; Krause-Jensen, Jakob; Saltofte, Margit

    2015-01-01

    . In this chapter, we argue that although anthropology has its specific methodology – including a myriad of ethnographic data-gathering tools, techniques, analytical approaches and theories – it must first and foremost be understood as a craft. Anthropology as craft requires a specific ‘anthropological sensibility......’ that differs from the standardized procedures of normal science. To establish our points we use an example of problem-based project work conducted by a group of Techno-Anthropology students at Aalborg University, we focus on key aspects of this craft and how the students began to learn it: For two weeks...... the students followed the work of a group of porters. Drawing on anthropological concepts and research strategies the students gained crucial insights about the potential effects of using tracking technologies in the hospital....

  3. Fibre tracking

    Gaillard, J.M.

    1994-03-01

    A large-size scintillating plastic fibre tracking detector was built as part of the upgrade of the UA2 central detector at the SPS proton-antiproton collider. The cylindrical fibre detector of average radius of 40 cm consisted of 60000 plastic fibres with an active length of 2.1 m. One of the main motivations was to improve the electron identification. The fibre ends were bunched to be coupled to read-out systems of image intensifier plus CCD, 32 in total. The quality and the reliability of the UA2 fibre detector performance exceeded expectations throughout its years of operation. A few examples of the use of image intensifiers and of scintillating fibres in biological instrumentation are described. (R.P.) 11 refs., 15 figs., 2 tabs

  4. SAW-Based Phononic Crystal Microfluidic Sensor-Microscale Realization of Velocimetry Approaches for Integrated Analytical Platform Applications.

    Oseev, Aleksandr; Lucklum, Ralf; Zubtsov, Mikhail; Schmidt, Marc-Peter; Mukhin, Nikolay V; Hirsch, Soeren

    2017-09-23

    The current work demonstrates a novel surface acoustic wave (SAW) based phononic crystal sensor approach that allows the integration of a velocimetry-based sensor concept into single chip integrated solutions, such as Lab-on-a-Chip devices. The introduced sensor platform merges advantages of ultrasonic velocimetry analytic systems and a microacoustic sensor approach. It is based on the analysis of structural resonances in a periodic composite arrangement of microfluidic channels confined within a liquid analyte. Completed theoretical and experimental investigations show the ability to utilize periodic structure localized modes for the detection of volumetric properties of liquids and prove the efficacy of the proposed sensor concept.

  5. Development of a radiation track structure clustering algorithm for the prediction of DNA DSB yields and radiation induced cell death in Eukaryotic cells.

    Douglass, Michael; Bezak, Eva; Penfold, Scott

    2015-04-21

    The preliminary framework of a combined radiobiological model is developed and calibrated in the current work. The model simulates the production of individual cells forming a tumour, the spatial distribution of individual ionization events (using Geant4-DNA) and the stochastic biochemical repair of DNA double strand breaks (DSBs) leading to the prediction of survival or death of individual cells. In the current work, we expand upon a previously developed tumour generation and irradiation model to include a stochastic ionization damage clustering and DNA lesion repair model. The Geant4 code enabled the positions of each ionization event in the cells to be simulated and recorded for analysis. An algorithm was developed to cluster the ionization events in each cell into simple and complex double strand breaks. The two lesion kinetic (TLK) model was then adapted to predict DSB repair kinetics and the resultant cell survival curve. The parameters in the cell survival model were then calibrated using experimental cell survival data of V79 cells after low energy proton irradiation. A monolayer of V79 cells was simulated using the tumour generation code developed previously. The cells were then irradiated by protons with mean energies of 0.76 MeV and 1.9 MeV using a customized version of Geant4. By replicating the experimental parameters of a low energy proton irradiation experiment and calibrating the model with two sets of data, the model is now capable of predicting V79 cell survival after low energy (cell survival probability, the cell survival probability is calculated for each cell in the geometric tumour model developed in the current work. This model uses fundamental measurable microscopic quantities such as genome length rather than macroscopic radiobiological quantities such as alpha/beta ratios. This means that the model can be theoretically used under a wide range of conditions with a single set of input parameters once calibrated for a given cell line.

  6. Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells

    Berry, David; Mader, Esther; Lee, Tae Kwon

    2015-01-01

    Microbial communities are essential to the function of virtually all ecosystems and eukaryotes, including humans. However, it is still a major challenge to identify microbial cells active under natural conditions in complex systems. In this study, we developed a new method to identify and sort ac...

  7. Tracking the stochastic fate of cells of the renin lineage after podocyte depletion using multicolor reporters and intravital imaging.

    Natalya V Kaverina

    Full Text Available Podocyte depletion plays a major role in focal segmental glomerular sclerosis (FSGS. Because cells of the renin lineage (CoRL serve as adult podocyte and parietal epithelial cell (PEC progenitor candidates, we generated Ren1cCre/R26R-ConfettiTG/WT and Ren1dCre/R26R-ConfettiTG/WT mice to determine CoRL clonality during podocyte replacement. Four CoRL reporters (GFP, YFP, RFP, CFP were restricted to cells in the juxtaglomerular compartment (JGC at baseline. Following abrupt podocyte depletion in experimental FSGS, all four CoRL reporters were detected in a subset of glomeruli at day 28, where they co-expressed de novo four podocyte proteins (podocin, nephrin, WT-1 and p57 and two glomerular parietal epithelial cell (PEC proteins (claudin-1, PAX8. To monitor the precise migration of a subset of CoRL over a 2w period following podocyte depletion, intravital multiphoton microscopy was used. Our findings demonstrate direct visual support for the migration of single CoRL from the JGC to the parietal Bowman's capsule, early proximal tubule, mesangium and glomerular tuft. In summary, these results suggest that following podocyte depletion, multi-clonal CoRL migrate to the glomerulus and replace podocyte and PECs in experimental FSGS.

  8. Assessing the accuracy of weather radar to track intense rain cells in the Greater Lyon area, France

    Renard, Florent; Chapon, Pierre-Marie; Comby, Jacques

    2012-01-01

    The Greater Lyon is a dense area located in the Rhône Valley in the south east of France. The conurbation counts 1.3 million inhabitants and the rainfall hazard is a great concern. However, until now, studies on rainfall over the Greater Lyon have only been based on the network of rain gauges, despite the presence of a C-band radar located in the close vicinity. Consequently, the first aim of this study was to investigate the hydrological quality of this radar. This assessment, based on comparison of radar estimations and rain-gauges values concludes that the radar data has overall a good quality since 2006. Given this good accuracy, this study made a next step and investigated the characteristics of intense rain cells that are responsible of the majority of floods in the Greater Lyon area. Improved knowledge on these rainfall cells is important to anticipate dangerous events and to improve the monitoring of the sewage system. This paper discusses the analysis of the ten most intense rainfall events in the 2001-2010 period. Spatial statistics pointed towards straight and linear movements of intense rainfall cells, independently on the ground surface conditions and the topography underneath. The speed of these cells was found nearly constant during a rainfall event, but depend from event to ranges on average from 25 to 66 km/h.

  9. EU PVSEC: New products: Cells and modules, production technology, monitoring and communication, measuring technology, inverters, tracking systems etc.; EU PVSEC: Produktneuheiten. Zellen und Module, Produktionstechnik, Monitoring und Kommunikation, Messtechnik, Wechselrichter, Nachfuehrsysteme, Was man sonst noch braucht

    Ossenbrink, Ralf; Augsten, Eva; Gesthuizen, Jan; Maeuler, Desiree; Buddensiek, Volker; Garus, Katharina

    2010-09-01

    The contribution takes a look back at the EU PVSEC trade fair and presents some of the innovations that were shown. There was news in many areas, e.g. cells and modules, production technology, monitoring and communication, measuring technology, inverters, tracking systems etc. (orig./AKB)

  10. Multimodal in vivo blood flow sensing combining particle image velocimetry and optical tweezers-based blood steering

    Meissner, Robert; Sugden, Wade W.; Siekmann, Arndt F.; Denz, Cornelia

    2018-02-01

    All higher developed organisms contain complex hierarchical networks of arteries, veins and capillaries. These constitute the cardiovascular system responsible for supplying nutrients, gas and waste exchange. Diseases related to the cardiovascular system are among the main causes for death worldwide. In order to understand the processes leading to arteriovenous malformation, we studied hereditary hemorrhagic telangiectasia (HHT), which has a prevalence of 1:5000 worldwide and causes internal bleeding. In zebrafish, HHT is induced by mutation of the endoglin gene involved in HHT and observed to reduce red blood cell (RBC) flow to intersegmental vessels (ISVs) in the tail due to malformations of the dorsal aorta (DA) and posterior cardinal vein (PCV). However, these capillaries are still functional. Changes in the blood flow pattern are observed from in vivo data from zebrafish embryos through particle image velocimetry (PIV). Wall shear rates (WSRs) and blood flow velocities are obtained non-invasively with millisecond resolution. We observe significant increases of blood flow velocity in the DA for endoglin-deficient zebrafish embryos (mutants) at 3 days post fertilization. In the PCV, this increase is even more pronounced. We identified an increased similarity between the DA and the PCV of mutant fish compared to siblings, i.e., unaffected fish. To counteract the reduced RBC flow to ISVs we implement optical tweezers (OT). RBCs are steered into previously unperfused ISVs showing a significant increase of RBC count per minute. We discuss limitations with respect to biocompatibility of optical tweezers in vivo and determination of in vivo wall shear stress (WSS) connected to normal and endoglin-deficicent zebrafish embryos.

  11. Tracking Boulders

    2006-01-01

    13 March 2006 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows a portion of a trough in the Sirenum Fossae region. On the floor and walls of the trough, large -- truck- to house-sized -- boulders are observed at rest. However, there is evidence in this image for the potential for mobility. In the central portion of the south (bottom) wall, a faint line of depressions extends from near the middle of the wall, down to the rippled trough floor, ending very near one of the many boulders in the area. This line of depressions is a boulder track; it indicates the path followed by the boulder as it trundled downslope and eventually came to rest on the trough floor. Because it is on Mars, even when the boulder is sitting still, this once-rolling stone gathers no moss. Location near: 29.4oS, 146.6oW Image width: 3 km (1.9 mi) Illumination from: upper left Season: Southern Summer

  12. INNER TRACKING

    P. Sharp

    The CMS Inner Tracking Detector continues to make good progress. The successful commissioning of ~ 25% of the Silicon Strip Tracker was completed in the Tracker Integration Facility (TIF) at CERN in July 2007 and the Tracker has since been prepared for moving and installation into CMS at P5. The Tracker was ready to move on schedule in September 2007. The Installation of the Tracker cooling pipes and LV cables between Patch Panel 1 (PP1) on the inside the CMS magnet cryostat, and the cooling plants and power system racks on the balconies has been completed. The optical fibres from PP1 to the readout FEDs in the USC have been installed, together with the Tracker cable channels, in parallel with the installation of the EB/HB services. All of the Tracker Safety, Power, DCS and the VME Readout Systems have been installed at P5 and are being tested and commissioned with CMS. It is planned to install the Tracker into CMS before Christmas. The Tracker will then be connected to the pre-installed services on Y...

  13. INNER TRACKING

    P. Sharp

    The CMS Inner Tracking Detector continues to make good progress. The successful commissioning of ~ 25% of the Silicon Strip Tracker was completed in the Tracker Integration Facility (TIF) at CERN on 18 July 2007 and the Tracker has since been prepared for moving and installation into CMS at P5. The Tracker will be ready to move on schedule in September 2007. The Installation of the Tracker cooling pipes and LV cables between Patch Panel 1 (PP1) on the inside the CMS magnet cryostat, and the cooling plants and power system racks on the balconies has been completed. The optical fibres from PP1 to the readout FEDs in the USC will be installed in parallel with the installation of the EB/HB services, and will be completed in October. It is planned to install the Tracker into CMS at the end of October, after the completion of the installation of the EB/HB services. The Tracker will then be connected to the pre-installed services on YB0 and commissioned with CMS in December. The FPix and BPix continue to make ...

  14. Tracking Electron Uptake from a Cathode into Shewanella Cells: Implications for Energy Acquisition from Solid-Substrate Electron Donors

    Annette R. Rowe

    2018-02-01

    Full Text Available While typically investigated as a microorganism capable of extracellular electron transfer to minerals or anodes, Shewanella oneidensis MR-1 can also facilitate electron flow from a cathode to terminal electron acceptors, such as fumarate or oxygen, thereby providing a model system for a process that has significant environmental and technological implications. This work demonstrates that cathodic electrons enter the electron transport chain of S. oneidensis when oxygen is used as the terminal electron acceptor. The effect of electron transport chain inhibitors suggested that a proton gradient is generated during cathode oxidation, consistent with the higher cellular ATP levels measured in cathode-respiring cells than in controls. Cathode oxidation also correlated with an increase in the cellular redox (NADH/FMNH2 pool determined with a bioluminescence assay, a proton uncoupler, and a mutant of proton-pumping NADH oxidase complex I. This work suggested that the generation of NADH/FMNH2 under cathodic conditions was linked to reverse electron flow mediated by complex I. A decrease in cathodic electron uptake was observed in various mutant strains, including those lacking the extracellular electron transfer components necessary for anodic-current generation. While no cell growth was observed under these conditions, here we show that cathode oxidation is linked to cellular energy acquisition, resulting in a quantifiable reduction in the cellular decay rate. This work highlights a potential mechanism for cell survival and/or persistence on cathodes, which might extend to environments where growth and division are severely limited.

  15. Tracking Electron Uptake from a Cathode into Shewanella Cells: Implications for Energy Acquisition from Solid-Substrate Electron Donors

    Rajeev, Pournami; Jain, Abhiney; Pirbadian, Sahand; Okamoto, Akihiro; Gralnick, Jeffrey A.; El-Naggar, Mohamed Y.; Nealson, Kenneth H.

    2018-01-01

    ABSTRACT While typically investigated as a microorganism capable of extracellular electron transfer to minerals or anodes, Shewanella oneidensis MR-1 can also facilitate electron flow from a cathode to terminal electron acceptors, such as fumarate or oxygen, thereby providing a model system for a process that has significant environmental and technological implications. This work demonstrates that cathodic electrons enter the electron transport chain of S. oneidensis when oxygen is used as the terminal electron acceptor. The effect of electron transport chain inhibitors suggested that a proton gradient is generated during cathode oxidation, consistent with the higher cellular ATP levels measured in cathode-respiring cells than in controls. Cathode oxidation also correlated with an increase in the cellular redox (NADH/FMNH2) pool determined with a bioluminescence assay, a proton uncoupler, and a mutant of proton-pumping NADH oxidase complex I. This work suggested that the generation of NADH/FMNH2 under cathodic conditions was linked to reverse electron flow mediated by complex I. A decrease in cathodic electron uptake was observed in various mutant strains, including those lacking the extracellular electron transfer components necessary for anodic-current generation. While no cell growth was observed under these conditions, here we show that cathode oxidation is linked to cellular energy acquisition, resulting in a quantifiable reduction in the cellular decay rate. This work highlights a potential mechanism for cell survival and/or persistence on cathodes, which might extend to environments where growth and division are severely limited. PMID:29487241

  16. Study of flow around model of cooling tower by means of 2D Particle Image Velocimetry measurement

    Barraclough, Veronika; Novotný, Jan; Šafařík, Pavel

    This paper deals with flow around a bluff body of hyperboloid shape. It combines results gathered in the course of research by means of Particle Image Velocimetry (PIV). The experiments were carried out by means of low-frequency 2D PIV and the Reynolds number was 43 000.

  17. Effects of Injection Timing on Fluid Flow Characteristics of Partially Premixed Combustion Based on High-Speed Particle Image Velocimetry

    Izadi Najafabadi, Mohammad; Tanov, Slavey; Wang, Hua; Somers, Bart; Johansson, Bengt; Dam, Nico

    2017-01-01

    behavior. The scope of the present study is to investigate the fluid flow characteristics of PPC at different injection timings. To this end, high-speed Particle Image Velocimetry (PIV) is implemented in a light-duty optical engine to measure fluid flow

  18. Acoustic radiation- and streaming-induced microparticle velocities determined by microparticle image velocimetry in an ultrasound symmetry plane

    Barnkob, Rune; Augustsson, Per; Laurell, Thomas

    2012-01-01

    We present microparticle image velocimetry measurements of suspended microparticles of diameters from 0.6 to 10μm undergoing acoustophoresis in an ultrasound symmetry plane in a microchannel. The motion of the smallest particles is dominated by the Stokes drag from the induced acoustic streaming...

  19. Selectivity and balance of spatial filtering velocimetry of objective speckles for measuring out-of-plane motion

    Jakobsen, Michael Linde; Yura, Hal T.; Hanson, Steen Grüner

    2015-01-01

    We probe the dynamics of objective laser speckles as the axial distance between the object and the observation plane changes. With the purpose of measuring out-of-plane motion in real time, we apply optical spatial filtering velocimetry to the speckle dynamics. To achieve this, a rotationally sym...

  20. Study of flow around model of cooling tower by means of 2D Particle Image Velocimetry measurement

    Barraclough Veronika

    2017-01-01

    Full Text Available This paper deals with flow around a bluff body of hyperboloid shape. It combines results gathered in the course of research by means of Particle Image Velocimetry (PIV. The experiments were carried out by means of low-frequency 2D PIV and the Reynolds number was 43 000.

  1. Prediction of production of {sup 22}Na in a gas-cell target irradiated by protons using Monte Carlo tracking

    Eslami, M., E-mail: mohammad.eslami25@yahoo.com [Department of Physics, Faculty of Science, University of Zanjan, Zengan (Zanjan) (Iran, Islamic Republic of); Kakavand, T. [Department of Physics, Faculty of Science, University of Zanjan, Zengan (Zanjan) (Iran, Islamic Republic of); Department of Physics, Faculty of Science, Imam Khomeini International University, Qazvin (Iran, Islamic Republic of); Mirzaii, M.; Rajabifar, S. [Agricultural, Medical and Industrial Research School, Nuclear Science and Technology Research Institute, AEOI, Karaj (Iran, Islamic Republic of)

    2015-01-01

    Highlights: • Angular distribution of the proton beam in a gaseous environment. • Particle energy distribution profile and proton flux within gas-cell target with MCNPX. • Detection of the residual nuclei during the nuclear reactions. • Estimation of production yield for {sup 22,nat}Ne(p,x){sup 22}Na reactions. - Abstract: The {sup 22}Ne(p,n){sup 22}Na is an optimal reaction for the cyclotron production of {sup 22}Na. This work tends to monitor the proton induced production of {sup 22}Na in a gas-cell target, containing natural and enriched neon gas, using Monte Carlo method. The excitation functions of reactions are calculated by both TALYS-1.6 and ALICE/ASH codes and then the optimum energy range of projectile for the high yield production is selected. A free gaseous environment of neon at a particular pressure and temperature is prearranged and the proton beam is transported within it using Monte Carlo codes MCNPX and SRIM. The beam monitoring performed by each of these codes indicates that the gas-cell has to be designed as conical frustum to reach desired interactions. The MCNPX is also employed to calculate the energy distribution of proton in the designed target and estimation of the residual nuclei during irradiation. The production yield of {sup 22}Na in {sup 22}Ne(p,n){sup 22}Na and {sup nat}Ne(p,x){sup 22}Na reactions are estimated and it shows a good agreement with the experimental results. The results demonstrate that Monte Carlo makes available a beneficial manner to design and optimize the gas targets as well as calibration of detectors, which can be used for the radionuclide production purposes.

  2. Ex-situ tracking solid oxide cell electrode microstructural evolution in a redox cycle by high resolution ptychographic nanotomography

    De Angelis, Salvatore; Jørgensen, Peter Stanley; Esposito, Vincenzo

    2017-01-01

    , the nickel and pore networks undergo major reorganization and the formation of internal voids is observed in the nickel-oxide particles after the oxidation. These observations are discussed in terms of reaction kinetics, electrode mechanical stress and the consequences of redox cycling on electrode...... towards this aim by visualizing a complete redox cycle in a solid oxide cell (SOC) electrode. The experiment demonstrates synchrotron-based ptychography as a method of imaging SOC electrodes, providing an unprecedented combination of 3D image quality and spatial resolution among non-destructive imaging...

  3. Estimation of the measurement uncertainty in magnetic resonance velocimetry based on statistical models

    Bruschewski, Martin; Schiffer, Heinz-Peter [Technische Universitaet Darmstadt, Institute of Gas Turbines and Aerospace Propulsion, Darmstadt (Germany); Freudenhammer, Daniel [Technische Universitaet Darmstadt, Institute of Fluid Mechanics and Aerodynamics, Center of Smart Interfaces, Darmstadt (Germany); Buchenberg, Waltraud B. [University Medical Center Freiburg, Medical Physics, Department of Radiology, Freiburg (Germany); Grundmann, Sven [University of Rostock, Institute of Fluid Mechanics, Rostock (Germany)

    2016-05-15

    Velocity measurements with magnetic resonance velocimetry offer outstanding possibilities for experimental fluid mechanics. The purpose of this study was to provide practical guidelines for the estimation of the measurement uncertainty in such experiments. Based on various test cases, it is shown that the uncertainty estimate can vary substantially depending on how the uncertainty is obtained. The conventional approach to estimate the uncertainty from the noise in the artifact-free background can lead to wrong results. A deviation of up to -75% is observed with the presented experiments. In addition, a similarly high deviation is demonstrated with the data from other studies. As a more accurate approach, the uncertainty is estimated directly from the image region with the flow sample. Two possible estimation methods are presented. (orig.)

  4. Estimation of the measurement uncertainty in magnetic resonance velocimetry based on statistical models

    Bruschewski, Martin; Freudenhammer, Daniel; Buchenberg, Waltraud B.; Schiffer, Heinz-Peter; Grundmann, Sven

    2016-05-01

    Velocity measurements with magnetic resonance velocimetry offer outstanding possibilities for experimental fluid mechanics. The purpose of this study was to provide practical guidelines for the estimation of the measurement uncertainty in such experiments. Based on various test cases, it is shown that the uncertainty estimate can vary substantially depending on how the uncertainty is obtained. The conventional approach to estimate the uncertainty from the noise in the artifact-free background can lead to wrong results. A deviation of up to -75 % is observed with the presented experiments. In addition, a similarly high deviation is demonstrated with the data from other studies. As a more accurate approach, the uncertainty is estimated directly from the image region with the flow sample. Two possible estimation methods are presented.

  5. Stereoscopic particle image velocimetry investigations of the mixed convection exchange flow through a horizontal vent

    Varrall, Kevin; Pretrel, Hugues; Vaux, Samuel; Vauquelin, Olivier

    2017-10-01

    The exchange flow through a horizontal vent linking two compartments (one above the other) is studied experimentally. This exchange is here governed by both the buoyant natural effect due to the temperature difference of the fluids in both compartments, and the effect of a (forced) mechanical ventilation applied in the lower compartment. Such a configuration leads to uni- or bi-directional flows through the vent. In the experiments, buoyancy is induced in the lower compartment thanks to an electrical resistor. The forced ventilation is applied in exhaust or supply modes and three different values of the vent area. To estimate both velocity fields and flow rates at the vent, measurements are realized at thermal steady state, flush the vent in the upper compartment using stereoscopic particle image velocimetry (SPIV), which is original for this kind of flow. The SPIV measurements allows the area occupied by both upward and downward flows to be determined.

  6. PIV-DCNN: cascaded deep convolutional neural networks for particle image velocimetry

    Lee, Yong; Yang, Hua; Yin, Zhouping

    2017-12-01

    Velocity estimation (extracting the displacement vector information) from the particle image pairs is of critical importance for particle image velocimetry. This problem is mostly transformed into finding the sub-pixel peak in a correlation map. To address the original displacement extraction problem, we propose a different evaluation scheme (PIV-DCNN) with four-level regression deep convolutional neural networks. At each level, the networks are trained to predict a vector from two input image patches. The low-level network is skilled at large displacement estimation and the high- level networks are devoted to improving the accuracy. Outlier replacement and symmetric window offset operation glue the well- functioning networks in a cascaded manner. Through comparison with the standard PIV methods (one-pass cross-correlation method, three-pass window deformation), the practicability of the proposed PIV-DCNN is verified by the application to a diversity of synthetic and experimental PIV images.

  7. Velocity Field Measurements of Human Coughing Using Time Resolved Particle Image Velocimetry

    Khan, T.; Marr, D. R.; Higuchi, H.; Glauser, M. N.

    2003-11-01

    Quantitative fluid mechanics analysis of human coughing has been carried out using new Time Resolved Particle Image Velocimetry (TRPIV). The study involves measurement of velocity vector time-histories and velocity profiles. It is focused on the average normal human coughing. Some work in the past on cough mechanics has involved measurement of flow rates, tidal volumes and sub-glottis pressure. However, data of unsteady velocity vector field of the exiting highly time-dependent jets is not available. In this study, human cough waveform data are first acquired in vivo using conventional respiratory instrumentation for various volunteers of different gender/age groups. The representative waveform is then reproduced with a coughing/breathing simulator (with or without a manikin) for TRPIV measurements and analysis. The results of this study would be useful not only for designing of indoor air quality and heating, ventilation and air conditioning systems, but also for devising means of protection against infectious diseases.

  8. Spatial filtering velocimetry revisited: exact short-time detecting schemes from arbitrarily small-size reticles

    Ando, S; Nara, T; Kurihara, T

    2014-01-01

    Spatial filtering velocimetry was proposed in 1963 by Ator as a velocity-sensing technique for aerial camera-control systems. The total intensity of a moving surface is observed through a set of parallel-slit reticles, resulting in a narrow-band temporal signal whose frequency is directly proportional to the image velocity. However, even despite its historical importance and inherent technical advantages, the mathematical formulation of this technique is only valid when infinite-length observation in both space and time is possible, which causes significant errors in most applications where a small receptive window and high resolution in both axes are desired. In this study, we apply a novel mathematical technique, the weighted integral method, to solve this problem, and obtain exact sensing schemes and algorithms for finite (arbitrarily small but non-zero) size reticles and short-time estimation. Practical considerations for utilizing these schemes are also explored both theoretically and experimentally. (paper)

  9. Correcting for color crosstalk and chromatic aberration in multicolor particle shadow velocimetry

    McPhail, M J; Fontaine, A A; Krane, M H; Goss, L; Crafton, J

    2015-01-01

    Color crosstalk and chromatic aberration can bias estimates of fluid velocity measured by color particle shadow velocimetry (CPSV), using multicolor illumination and a color camera. This article describes corrections to remove these bias errors, and their evaluation. Color crosstalk removal is demonstrated with linear unmixing. It is also shown that chromatic aberrations may be removed using either scale calibration, or by processing an image illuminated by all colors simultaneously. CPSV measurements of a fully developed turbulent pipe flow of glycerin were conducted. Corrected velocity statistics from these measurements were compared to both single-color PSV and LDV measurements and showed excellent agreement to fourth-order, to well into the viscous sublayer. Recommendations for practical assessment and correction of color aberration and color crosstalk are discussed. (paper)

  10. Investigating fundamental properties of wind turbine wake structure using particle image velocimetry

    Whale, J. [Univ. of Edinburgh, Dept. of Mechanical Engineering, Edinburgh (United Kingdom)

    1997-08-01

    Low Reynolds number flow visualization tests are often used for showing the flow pattern changes associated with changes in lift-coefficients at a higher Reynolds number. In wind turbine studies, analysis of measured wake structures at small scale may reveal fundamental properties of the wake which will offer wake modellers a more complete understanding of rotor flows. Measurements are presented from experiments on a model wind turbine rig conducted in a water channel. The laser-optics technique of Particle Image Velocimetry (PIV) is used to make simultaneous multi-point measurements of the wake flow behind small-scale rotors. Analysis of the PIV data shows trends in velocity and vorticity structure in the wake. Study of the flow close to the rotor plane reveals information on stalled flow and blade performance. (au)

  11. Study of fish response using particle image velocimetry and high-speed, high-resolution imaging

    Deng, Z. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Richmond, M. C. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Mueller, R. P. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Gruensch, G. R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2004-10-01

    Fish swimming has fascinated both engineers and fish biologists for decades. Digital particle image velocimetry (DPIV) and high-speed, high-resolution digital imaging are recently developed analysis tools that can help engineers and biologists better understand how fish respond to turbulent environments. This report details studies to evaluate DPIV. The studies included a review of existing literature on DPIV, preliminary studies to test the feasibility of using DPIV conducted at our Flow Biology Laboratory in Richland, Washington September through December 2003, and applications of high-speed, high-resolution digital imaging with advanced motion analysis to investigations of fish injury mechanisms in turbulent shear flows and bead trajectories in laboratory physical models. Several conclusions were drawn based on these studies, which are summarized as recommendations for proposed research at the end of this report.

  12. Speckle and fringe dynamics in imagingspeckle-pattern interferometry for spatial-filtering velocimetry

    Jakobsen, Michael Linde; Iversen, Theis F. Q.; Yura, Harold T.

    2011-01-01

    This paper analyzes the dynamics of laser speckles and fringes, formed in an imaging-speckle-pattern interferometer with the purpose of sensing linear three-dimensional motion and out-of-plane components of rotation in real time, using optical spatial-filtering-velocimetry techniques. The ensemble......-average definition of the cross-correlation function is applied to the intensity distributions, obtained in the observation plane at two positions of the object. The theoretical analysis provides a description for the dynamics of both the speckles and the fringes. The analysis reveals that both the magnitude...... and direction of all three linear displacement components of the object movement can be determined. Simultaneously, out-ofplane rotation of the object including the corresponding directions can be determined from the spatial gradient of the in-plane fringe motion throughout the observation plane. The theory...

  13. Shock Initiation of Wedge-shaped Explosive Measured with Smear Camera and Photon Doppler Velocimetry

    Gu, Yan

    2017-06-01

    Triaminotrinitrobenzene (TATB) is an important insensitive high explosive in conventional weapons due to its safety and high energy. In order to have an insight into the shock initiation performance of a TATB-based insensitive high explosive (IHE), experimental measurements of the particle velocity histories of the TATB-based Explosive using Photon Doppler Velocimetry and shock wave profile of the TATB-based explosive using High Speed Rotating Mirror Smear Camera had been performed. In this paper, we would describe the shock initiation performance of the TATB-based explosive by run-to-detonation distance and the particle velocity history at an initialization shock of about 7.9 GPa. The parameters of hugoniot of unreacted the TATB-based explosive and Pop relationship could be derived with the particle velocity history obtained in this paper.

  14. X-ray doppler velocimetry for diagnosis of fluid motion in ICF implosions

    Koch, J. A.; King, J. A.; Huffman, E.; Freeman, R. R.; Dutra, E. C.; Field, J. E.; Kilkenny, J. D.; Hall, G. N.; Harding, E.; Rochau, G. A.; Porter, J. L.; Covington, A. M.; Beg, F. N.

    2017-08-01

    We are developing a novel diagnostic for measurement of bulk fluid motion in materials, that is particularly applicable to very hot, x-ray emitting plasmas in the High Energy Density Physics (HEDP) regime. The X-ray Doppler Velocimetry (XDV) technique relies on monochromatic imaging in multiple x-ray energy bands near the center of an x-ray emission line in a plasma, and utilizes bent imaging crystals. Higher energy bands are preferentially sensitive to plasma moving towards the viewer, while lower energy bands are preferentially sensitive to plasma moving away from the viewer. Combining multiple images in different energy bands allows for a reconstruction of the fluid velocity field integrated along the line of sight. We review the technique, and we discuss progress towards benchmarking the technique with proof-of-principle HEDP experiments.

  15. Spatial filtering velocimetry for real-time out-of-plane displacement measurements

    Olesen, Anders Sig; Yura, H.T.; Jakobsen, Michael Linde

    2016-01-01

    power spectrum of the photocurrent produced by this filter. This main contribution of this paper is a model, which describe the selectivity of the sensor, applied to speckle dynamics generated by an object moving out-of-plane. To motivate our interest in these filters we also present an all optical......We probe the dynamics of objective laser speckles as the axial distance between the object and the observation plane changes. With the purpose of measuring out-of-plane motion in real time, we apply optical spatial filtering velocimetry to the speckle dynamics. To achieve this, a rotationally...... symmetric spatial filter is designed. The spatial filter converts the speckle dynamics into a photocurrent with a quasi-sinusoidal response to the out-of-plane motion. The selectivity of the sensor relates directly to the uncertainty on sensor measurements. The selectivity most be derived from a temporal...

  16. Spectroscopic interpretation and velocimetry analysis of fluctuations in a cylindrical plasma recorded by a fast camera

    Oldenbürger, S.; Brandt, C.; Brochard, F.; Lemoine, N.; Bonhomme, G.

    2010-06-01

    Fast visible imaging is used on a cylindrical magnetized argon plasma produced by thermionic discharge in the Mirabelle device. To link the information collected with the camera to a physical quantity, fast camera movies of plasma structures are compared to Langmuir probe measurements. High correlation is found between light fluctuations and plasma density fluctuations. Contributions from neutral argon and ionized argon to the overall light intensity are separated by using interference filters and a light intensifier. Light emitting transitions are shown to involve a metastable neutral argon state that can be excited by thermal plasma electrons, thus explaining the good correlation between light and density fluctuations. The propagation velocity of plasma structures is calculated by adapting velocimetry methods to the fast camera movies. The resulting estimates of instantaneous propagation velocity are in agreement with former experiments. The computation of mean velocities is discussed.

  17. Spectroscopic interpretation and velocimetry analysis of fluctuations in a cylindrical plasma recorded by a fast camera

    Oldenbuerger, S.; Brandt, C.; Brochard, F.; Lemoine, N.; Bonhomme, G.

    2010-01-01

    Fast visible imaging is used on a cylindrical magnetized argon plasma produced by thermionic discharge in the Mirabelle device. To link the information collected with the camera to a physical quantity, fast camera movies of plasma structures are compared to Langmuir probe measurements. High correlation is found between light fluctuations and plasma density fluctuations. Contributions from neutral argon and ionized argon to the overall light intensity are separated by using interference filters and a light intensifier. Light emitting transitions are shown to involve a metastable neutral argon state that can be excited by thermal plasma electrons, thus explaining the good correlation between light and density fluctuations. The propagation velocity of plasma structures is calculated by adapting velocimetry methods to the fast camera movies. The resulting estimates of instantaneous propagation velocity are in agreement with former experiments. The computation of mean velocities is discussed.

  18. Two-phase velocity measurements around cylinders using particle image velocimetry

    Hassan, Y.A.; Philip, O.G.; Schmidl, W.D. [Texas A& M Univ., College Station, TX (United States)] [and others

    1995-09-01

    The particle Image Velocimetry flow measurement technique was used to study both single-phase flow and two-phase flow across a cylindrical rod inserted in a channel. First, a flow consisting of only a single-phase fluid was studied. The experiment consisted of running a laminar flow over four rods inserted in a channel. The water flow rate was 126 cm{sup 3}/s. Then a two-phase flow was studied. A mixture of water and small air bubbles was used. The water flow rate was 378 cm{sup 3}/s and the air flow rate was approximately 30 cm{sup 3}/s. The data are analyzed to obtain the velocity fields for both experiments. After interpretation of the velocity data, forces acting on a bubble entrained by the vortex were calculated successfully. The lift and drag coefficients were calculated using the velocity measurements and the force data.

  19. Particle image velocimetry investigation of flow over unsteady airfoil with trailing-edge strip

    Gerontakos, P.; Lee, T. [McGill University, Montreal, QC (Canada)

    2008-04-15

    The flow over a flapped NACA 0012 airfoil, oscillated slightly through the static-stall angle, was investigated by using particle image velocimetry, and was supplemented by surface pressure and dynamic-load measurements. A significant increase in the dynamic lift force and nose-down pitching moment was observed. The most pronounced flow phenomenon was the formation and detachment of an energetic leading-edge vortex compared to the no-flapped airfoil. The details of the underlying physical mechanisms responsible for the various light-stall flow processes were provided via the instantaneous velocity and vorticity fields measurements. In contrast to the Gurney flap, the inverted trailing-edge strip led to an improved negative damping while a reduced lift force. The addition of an inverted strip always led to the appearance of a Karman-type vortex shedding street immediately downstream of the strip over the entire oscillation cycle. (orig.)

  20. Rainbow Particle Imaging Velocimetry for Dense 3D Fluid Velocity Imaging

    Xiong, Jinhui

    2017-04-11

    Despite significant recent progress, dense, time-resolved imaging of complex, non-stationary 3D flow velocities remains an elusive goal. In this work we tackle this problem by extending an established 2D method, Particle Imaging Velocimetry, to three dimensions by encoding depth into color. The encoding is achieved by illuminating the flow volume with a continuum of light planes (a “rainbow”), such that each depth corresponds to a specific wavelength of light. A diffractive component in the camera optics ensures that all planes are in focus simultaneously. For reconstruction, we derive an image formation model for recovering stationary 3D particle positions. 3D velocity estimation is achieved with a variant of 3D optical flow that accounts for both physical constraints as well as the rainbow image formation model. We evaluate our method with both simulations and an experimental prototype setup.

  1. Particle image and acoustic Doppler velocimetry analysis of a cross-flow turbine wake

    Strom, Benjamin; Brunton, Steven; Polagye, Brian

    2017-11-01

    Cross-flow turbines have advantageous properties for converting kinetic energy in wind and water currents to rotational mechanical energy and subsequently electrical power. A thorough understanding of cross-flow turbine wakes aids understanding of rotor flow physics, assists geometric array design, and informs control strategies for individual turbines in arrays. In this work, the wake physics of a scale model cross-flow turbine are investigated experimentally. Three-component velocity measurements are taken downstream of a two-bladed turbine in a recirculating water channel. Time-resolved stereoscopic particle image and acoustic Doppler velocimetry are compared for planes normal to and distributed along the turbine rotational axis. Wake features are described using proper orthogonal decomposition, dynamic mode decomposition, and the finite-time Lyapunov exponent. Consequences for downstream turbine placement are discussed in conjunction with two-turbine array experiments.

  2. Stereo particle image velocimetry set up for measurements in the wake of scaled wind turbines

    Campanardi, Gabriele; Grassi, Donato; Zanotti, Alex; Nanos, Emmanouil M.; Campagnolo, Filippo; Croce, Alessandro; Bottasso, Carlo L.

    2017-08-01

    Stereo particle image velocimetry measurements were carried out in the boundary layer test section of Politecnico di Milano large wind tunnel to survey the wake of a scaled wind turbine model designed and developed by Technische Universität München. The stereo PIV instrumentation was set up to survey the three velocity components on cross-flow planes at different longitudinal locations. The area of investigation covered the entire extent of the wind turbines wake that was scanned by the use of two separate traversing systems for both the laser and the cameras. Such instrumentation set up enabled to gain rapidly high quality results suitable to characterise the behaviour of the flow field in the wake of the scaled wind turbine. This would be very useful for the evaluation of the performance of wind farm control methodologies based on wake redirection and for the validation of CFD tools.

  3. Functionalised alginate flow seeding microparticles for use in Particle Image Velocimetry (PIV).

    Varela, Sylvana; Balagué, Isaac; Sancho, Irene; Ertürk, Nihal; Ferrando, Montserrat; Vernet, Anton

    2016-01-01

    Alginate microparticles as flow seeding fulfil all the requirements that are recommended for the velocity measurements in Particle Image Velocimetry (PIV). These spherical microparticles offer the advantage of being environmentally friendly, having excellent seeding properties and they can be produced via a very simple process. In the present study, the performances of alginate microparticles functionalised with a fluorescent dye, Rhodamine B (RhB), for PIV have been studied. The efficacy of fluorescence is appreciated in a number of PIV applications since it can boost the signal-to-noise ratio. Alginate microparticles functionalised with RhB have high emission efficiency, desirable match with fluid density and controlled size. The study of the particles behaviour in strong acid and basic solutions and ammonia is also included. This type of particles can be used for measurements with PIV and Planar Laser Induced Fluorescence (PLIF) simultaneously, including acid-base reactions.

  4. Acceleration Characteristics of a Rock Slide Using the Particle Image Velocimetry Technique

    Guoqing Chen

    2016-01-01

    Full Text Available The Particle Image Velocimetry (PIV technique with high precision and spatial resolution is a suitable sensor for flow field experiments. In this paper, the PIV technology was used to monitor the development of a displacement field, velocity field and acceleration field of a rock slide. It was found that the peak acceleration of the sliding surface appeared earlier than the peak acceleration of the sliding body. The characteristics of the rock slide including the short failure time, high velocities, and large accelerations indicate that the sliding forces and energy release rate of the slope are high. The deformation field showed that the sliding body was sliding outwards along the sliding surface while the sliding bed moved in an opposite direction. Moving upwards at the top of the sliding bed can be one of the warning signs for rock slide failure.

  5. Measurement of fluid velocity development behind a circular cylinder using particle image velocimetry (PIV)

    Goharzadeh, Afshin; Molki, Arman

    2015-01-01

    In this paper we present a non-intrusive experimental approach for obtaining a two-dimensional velocity distribution around a 22 mm diameter circular cylinder mounted in a water tunnel. Measurements were performed for a constant Reynolds number of 7670 using a commercial standard particle image velocimetry (PIV) system. Different flow patterns generated behind the circular cylinder are discussed. Both instantaneous and time-averaged velocity distributions with corresponding streamlines are obtained. Key concepts in fluid mechanics, such as contra-rotating vortices, von Kármán vortex street, and laminar-turbulent flow, are discussed. In addition, brief historical information pertaining to the development of flow measurement techniques—in particular, PIV—is described. (paper)

  6. High resolution measurement of the velocity profiles of channel flows using the particle image velocimetry technique

    Nor Azizi Mohamed

    2000-01-01

    The high resolution velocity profiles of a uniform steady channel flow and a flow beneath waves were obtained using the particle image velocimetry (PIV) technique. The velocity profiles for each flow were calculated for both components. It is shown that the profiles obtained are very precise, displaying the point velocities from a few millimeters from the bottom of the channel up to the water surface across the water depth. In the case of the wave-induced flow, the profiles are shown under the respective wave phases and given in a plane representation. High resolution measurement of point velocities in a flow is achievable using PIV and invaluable when applied to a complex flow. (Author)

  7. Application of FLEET Velocimetry in the NASA Langley 0.3-meter Transonic Cryogenic Tunnel

    Burns, Ross A.; Danehy, Paul M.; Halls, Benjamin R.; Jiang, Naibo

    2015-01-01

    Femtosecond laser electronic excitation and tagging (FLEET) velocimetry is demonstrated in a large-scale transonic cryogenic wind tunnel. Test conditions include total pressures, total temperatures, and Mach numbers ranging from 15 to 58 psia, 200 to 295 K, and 0.2 to 0.75, respectively. Freestream velocity measurements exhibit accuracies within 1 percent and precisions better than 1 m/s. The measured velocities adhere closely to isentropic flow theory over the domain of temperatures and pressures that were tested. Additional velocity measurements are made within the tunnel boundary layer; virtual trajectories traced out by the FLEET signal are indicative of the characteristic turbulent behavior in this region of the flow, where the unsteadiness increases demonstrably as the wall is approached. Mean velocities taken within the boundary layer are in agreement with theoretical velocity profiles, though the fluctuating velocities exhibit a greater deviation from theoretical predictions.

  8. Shifted knife-edge aperture digital in-line holography for fluid velocimetry.

    Palero, Virginia; Lobera, Julia; Andrés, Nieves; Arroyo, M Pilar

    2014-06-01

    We describe a digital holography technique that, with the simplicity of an in-line configuration, produces holograms where the real and virtual images are completely separated, as in an off-axis configuration. An in-line setup, in which the object is imaged near the sensor, is modified by placing a shifted knife-edge aperture that blocks half the frequency spectrum at the focal plane of the imaging lens. This simple modification of the in-line holographic configuration allows discriminating the virtual and real images. As a fluid velocimetry technique, the use of this aperture removes the minimum defocusing distance requisite and reduces the out-of-plane velocity measurement errors of classical in-line holography. Results with different test objects are shown.

  9. Real-time tracking of hydrogen peroxide secreted by live cells using MnO{sub 2} nanoparticles intercalated layered doubled hydroxide nanohybrids

    Asif, Muhammad; Aziz, Ayesha [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, Hubei Key Laboratory of Material Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, 430074 (China); Dao, Anh Quang [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, Hubei Key Laboratory of Material Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, 430074 (China); Hue Industrial College, 70 Nguyen Hue, Hue, Thua Thien Hue, 531081 (Viet Nam); Hakeem, Abdul; Wang, Haitao; Dong, Shuang; Zhang, Guoan [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, Hubei Key Laboratory of Material Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, 430074 (China); Xiao, Fei [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, Hubei Key Laboratory of Material Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, 430074 (China); Shenzhen Institute of Huazhong University of Science & Technology, Shenzhen, 518000 (China); Liu, Hongfang, E-mail: liuhf@hust.edu.cn [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, Hubei Key Laboratory of Material Chemistry and Service Failure, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, 430074 (China); Shenzhen Institute of Huazhong University of Science & Technology, Shenzhen, 518000 (China)

    2015-10-22

    We report a facile and green method for the fabrication of new type of electrocatalysts based on MnO{sub 2} nanoparticles incorporated on MgAl LDH P-type semiconductive channel and explore its practical applications as high-performance electrode materials for electrochemical biosensor. A series of MgAl layered doubled hydroxide (LDH) nanohybrids with fixed Mg/Al (M{sup 2+}/M{sup 3+} atomic ratio of 3) and varied amount of MnCl{sub 2}.4H{sub 2}O are fabricated by a facile co-precipitation method. This approach demonstrates the combination of distinct properties including excellent intercalation features of LDH for entrapping nanoparticles and high loading of MnO{sub 2} nanoparticles in the host layers of LDH. Among all samples, Mn5–MgAl with 0.04% loaded manganese has a good crystalline morphology. A well-dispersed MnO{sub 2} nanoparticles encapsulated into the host matrix of hydrotalcite exhibit enhanced electrocatalytic activity towards the reduction of H{sub 2}O{sub 2} as well as excellent stability, selectivity and reproducibility due to synergistic effect of good catalytic ability of MnO{sub 2} and conductive MgAl LDH. Glass carbon electrode (GCE) modified with Mn5–MgAl possesses a wide linear range of 0.05–78 mM, lowest detection limit 5 μM (S/N = 3) and detection sensitivity of 0.9352 μAmM{sup −1}. This outstanding performance enables it to be used for real-time tracking of H{sub 2}O{sub 2} secreted by live HeLa cells. This work may provide new insight in clinical diagnosis, on-site environmental analysis and point of care testing devices. - Highlights: • MnO{sub 2}MgAl nanohybrids have been fabricated by a facile and robust co-precipitation approach. • MgAl layered doubled hydroxide can be used for the intercalation of MnO{sub 2} nanoparticles. • MgAl layered doubled hydroxide nanohybrid serves as p-type semiconductive channel for efficient electrocatalysis. • The nanohybrid electrode demonstrates excellent electrochemical performance

  10. Particle image velocimetry measurements of 2-dimensional velocity field around twisted tape

    Song, Min Seop; Park, So Hyun; Kim, Eung Soo, E-mail: kes7741@snu.ac.kr

    2016-11-01

    Highlights: • Measurements of the flow field in a pipe with twisted tape were conducted by particle image velocimetry (PIV). • A novel matching index of refraction technique utilizing 3D printing and oil mixture was adopted to make the test section transparent. • Undistorted particle images were clearly captured in the presence of twisted tape. • 2D flow field in the pipe with twisted tape revealed the characteristic two-peak velocity profile. - Abstract: Twisted tape is a passive component used to enhance heat exchange in various devices. It induces swirl flow that increases the mixing of fluid. Thus, ITER selected the twisted tape as one of the candidates for turbulence promoting in the divertor cooling. Previous study was mainly focused on the thermohydraulic performance of the twisted tape. As detailed data on the velocity field around the twisted tape was insufficient, flow visualization study was performed to provide fundamental data on velocity field. To visualize the flow in a complex structure, novel matching index of refraction technique was used with 3-D printing and mixture of anise and mineral oil. This technique enables the camera to capture undistorted particle image for velocity field measurement. Velocity fields at Reynolds number 1370–9591 for 3 different measurement plane were obtained through particle image velocimetry. The 2-dimensional averaged velocity field data were obtained from 177 pair of instantaneous velocity fields. It reveals the characteristic two-peak flow motion in axial direction. In addition, the normalized velocity profiles were converged with increase of Reynolds numbers. Finally, the uncertainty of the result data was analyzed.

  11. Particle image velocimetry measurements of Mach 3 turbulent boundary layers at low Reynolds numbers

    Brooks, J. M.; Gupta, A. K.; Smith, M. S.; Marineau, E. C.

    2018-05-01

    Particle image velocimetry (PIV) measurements of Mach 3 turbulent boundary layers (TBL) have been performed under low Reynolds number conditions, Re_τ =200{-}1000, typical of direct numerical simulations (DNS). Three reservoir pressures and three measurement locations create an overlap in parameter space at one research facility. This allows us to assess the effects of Reynolds number, particle response and boundary layer thickness separate from facility specific experimental apparatus or methods. The Morkovin-scaled streamwise fluctuating velocity profiles agree well with published experimental and numerical data and show a small standard deviation among the nine test conditions. The wall-normal fluctuating velocity profiles show larger variations which appears to be due to particle lag. Prior to the current study, no detailed experimental study characterizing the effect of Stokes number on attenuating wall-normal fluctuating velocities has been performed. A linear variation is found between the Stokes number ( St) and the relative error in wall-normal fluctuating velocity magnitude (compared to hot wire anemometry data from Klebanoff, Characteristics of Turbulence in a Boundary Layer with Zero Pressure Gradient. Tech. Rep. NACA-TR-1247, National Advisory Committee for Aeronautics, Springfield, Virginia, 1955). The relative error ranges from about 10% for St=0.26 to over 50% for St=1.06. Particle lag and spatial resolution are shown to act as low-pass filters on the fluctuating velocity power spectral densities which limit the measurable energy content. The wall-normal component appears more susceptible to these effects due to the flatter spectrum profile which indicates that there is additional energy at higher wave numbers not measured by PIV. The upstream inclination and spatial correlation extent of coherent turbulent structures agree well with published data including those using krypton tagging velocimetry (KTV) performed at the same facility.

  12. Tracking of Neural Stem Cells in Rats with Intracerebral Hemorrhage by the Use of 3T MRI

    Chang, Nam Kyu; Jeong, Yong Yeon; Park, Jong Seong

    2008-01-01

    To access the feasibility of clinically available 3T MRI to detect the migration of labeled neural stem cells (NSCs) in intracerebral hemorrhage (ICH) in a rat model. The ethics committee of our institution approved this study. ICH was induced by the injection of collagenase type IV into the right striatum of ten Sprague-Dawley rats. Human NSCs conjugated with Feridex (superparamagnetic iron oxide: SPIO) were transplanted into the left striatum one week after ICH induction. MRI was performed on a 3T scanner during the first, second, third, fourth, and sixth weeks post-transplantation. MRI was obtained using coronal T2- and T2 * -weighted sequences. Two rats were sacrificed every week after in vivo MRI in order to analyze the histological findings. ICH in the right striatum was detected by MRI one and two weeks after transplantation without migration of the NSCs. There was no migration of the NSCs as seen on the histological findings one week after transplantation. The histological findings two weeks after transplantation showed a small number of NSCs along the corpus callosum. On MRI three weeks after transplantation, there was a hypointense line along the corpus callosum and decreased signal intensity in the right periventricular region. Histological findings three weeks after transplantation confirmed the presence of the hypointense line representing SPIO-labeled NSCs. MRI four and six weeks after transplantation showed a hypointense spot in the right periventricular region. The histological findings four and six weeks after transplantation showed the presence of prominent NSCs in the right periventricular region. 3T MRI can detect the migration of NSCs in rats with ICH along the corpus callosum. Therefore, 3T MRI could be feasible for detecting the migration of NSCs in the clinical setting of stem cell therapy

  13. Multiple objects tracking in fluorescence microscopy.

    Kalaidzidis, Yannis

    2009-01-01

    Many processes in cell biology are connected to the movement of compact entities: intracellular vesicles and even single molecules. The tracking of individual objects is important for understanding cellular dynamics. Here we describe the tracking algorithms which have been developed in the non-biological fields and successfully applied to object detection and tracking in biological applications. The characteristics features of the different algorithms are compared.

  14. Characterizing the DNA Damage Response by Cell Tracking Algorithms and Cell Features Classification Using High-Content Time-Lapse Analysis.

    Walter Georgescu

    Full Text Available Traditionally, the kinetics of DNA repair have been estimated using immunocytochemistry by labeling proteins involved in the DNA damage response (DDR with fluorescent markers in a fixed cell assay. However, detailed knowledge of DDR dynamics across multiple cell generations cannot be obtained using a limited number of fixed cell time-points. Here we report on the dynamics of 53BP1 radiation induced foci (RIF across multiple cell generations using live cell imaging of non-malignant human mammary epithelial cells (MCF10A expressing histone H2B-GFP and the DNA repair protein 53BP1-mCherry. Using automatic extraction of RIF imaging features and linear programming techniques, we were able to characterize detailed RIF kinetics for 24 hours before and 24 hours after exposure to low and high doses of ionizing radiation. High-content-analysis at the single cell level over hundreds of cells allows us to quantify precisely the dose dependence of 53BP1 protein production, RIF nuclear localization and RIF movement after exposure to X-ray. Using elastic registration techniques based on the nuclear pattern of individual cells, we could describe the motion of individual RIF precisely within the nucleus. We show that DNA repair occurs in a limited number of large domains, within which multiple small RIFs form, merge and/or resolve with random motion following normal diffusion law. Large foci formation is shown to be mainly happening through the merging of smaller RIF rather than through growth of an individual focus. We estimate repair domain sizes of 7.5 to 11 µm2 with a maximum number of ~15 domains per MCF10A cell. This work also highlights DDR which are specific to doses larger than 1 Gy such as rapid 53BP1 protein increase in the nucleus and foci diffusion rates that are significantly faster than for spontaneous foci movement. We hypothesize that RIF merging reflects a "stressed" DNA repair process that has been taken outside physiological conditions when

  15. Dynamic kirigami structures for integrated solar tracking

    Lamoureux, Aaron; Lee, Kyusang; Shlian, Matthew; Forrest, Stephen R.; Shtein, Max

    2015-01-01

    Optical tracking is often combined with conventional flat panel solar cells to maximize electrical power generation over the course of a day. However, conventional trackers are complex and often require costly and cumbersome structural components to support system weight. Here we use kirigami (the art of paper cutting) to realize novel solar cells where tracking is integral to the structure at the substrate level. Specifically, an elegant cut pattern is made in thin-film gallium arsenide solar cells, which are then stretched to produce an array of tilted surface elements which can be controlled to within ±1°. We analyze the combined optical and mechanical properties of the tracking system, and demonstrate a mechanically robust system with optical tracking efficiencies matching conventional trackers. This design suggests a pathway towards enabling new applications for solar tracking, as well as inspiring a broader range of optoelectronic and mechanical devices. PMID:26348820

  16. Identification of core components and transient interactors of the peroxisomal importomer by dual-track stable isotope labeling with amino acids in cell culture analysis.

    Oeljeklaus, Silke; Reinartz, Benedikt S; Wolf, Janina; Wiese, Sebastian; Tonillo, Jason; Podwojski, Katharina; Kuhlmann, Katja; Stephan, Christian; Meyer, Helmut E; Schliebs, Wolfgang; Brocard, Cécile; Erdmann, Ralf; Warscheid, Bettina

    2012-04-06

    The importomer complex plays an essential role in the biogenesis of peroxisomes by mediating the translocation of matrix proteins across the organellar membrane. A central part of this highly dynamic import machinery is the docking complex consisting of Pex14p, Pex13p, and Pex17p that is linked to the RING finger complex (Pex2p, Pex10p, Pex12p) via Pex8p. To gain detailed knowledge on the molecular players governing peroxisomal matrix protein import and, thus, the integrity and functionality of peroxisomes, we aimed at a most comprehensive investigation of stable and transient interaction partners of Pex14p, the central component of the importomer. To this end, we performed a thorough quantitative proteomics study based on epitope tagging of Pex14p combined with dual-track stable isotope labeling with amino acids in cell culture-mass spectrometry (SILAC-MS) analysis of affinity-purified Pex14p complexes and statistics. The results led to the establishment of the so far most extensive Pex14p interactome, comprising 9 core and further 12 transient components. We confirmed virtually all known Pex14p interaction partners including the core constituents of the importomer as well as Pex5p, Pex11p, Pex15p, and Dyn2p. More importantly, we identified new transient interaction partners (Pex25p, Hrr25p, Esl2p, prohibitin) that provide a valuable resource for future investigations on the functionality, dynamics, and regulation of the peroxisomal importomer.

  17. Solar tracking system

    Okandan, Murat; Nielson, Gregory N.

    2016-07-12

    Solar tracking systems, as well as methods of using such solar tracking systems, are disclosed. More particularly, embodiments of the solar tracking systems include lateral supports horizontally positioned between uprights to support photovoltaic modules. The lateral supports may be raised and lowered along the uprights or translated to cause the photovoltaic modules to track the moving sun.

  18. Central and forward tracking collaboration

    Foster, R.; Hanson, G.; Luehring, F.; Luo, X.; Martin, B.; Ogren, H.; Rust, D.R.; Wente, E.; Adrian, B.; Alexander, D.; Ells, F.; Erdos, E.; Ford, W.T.; Johnson, D.; Lohner, M.; Rankin, P.; Schultz, G.; Newcomer, F.M.; Van Berg, R.; Williams, H.H.; Arai, Y.; Hess, D.; Kadyk, J.A.; Palounek, A.P.T.; Wise, J.; Chapman, J.W.; Dunn, A.; Edwards, M.; Hiddleston, J.W.; Payne, B.T.; Amery, C.A.; Bailey, J.M.; Dainton, J.B.; Gabathuler, E.; Maxfield, S.J.; Morton, J.M.; Muir, A.; Patel, G.D.; Sanders, P.; Raine, C.; Saxon, D.H.; Hackworth, D.T.; Swensrud, R.L.; Newfield, S.; Sadler, C.; Va'vra, J.

    1991-01-01

    The goal of this subsystem R ampersand D project is to carry out a detailed study and design of a complete wire chamber tracking system covering pseudorapidity |η| ≤ 2.5 in a solenoidal detector for the SSC. Most of our group are now part of the Solenoidal Detector Collaboration (SDC), so the work has evolved into developing a tracking system conceptual design for the SDC detector. The design discussed in this report uses straw tube drift chambers for the central tracking region. Because of the high rates in the SSC environment, a small cell design is needed for wire chambers in the central region. Straw tubes as small cells offer many advantages because the sense wire is enclosed in a continuous cathode, and the wire tension due to the sense wire only can be supported without a massive structure. The straw tubes are grouped together to form superlayers in order to provide local track segments. The superlayers are composed of modules consisting of about two hundred straw tubes enclosed in a carbon fiber composite shell. Straw tubes have been used in previous experiments for small vertex drift chambers. However, they have never before been used for a large tracking system

  19. A PTV method based on ultrasound imaging and feature tracking in a low-concentration sediment-laden flow

    Ma, Zhimin; Hu, Wenbin; Zhao, Xiaohong; Tao, Weiliang

    2018-02-01

    This study aims to provide a particle tracking velocimetry (PTV) method based on ultrasound imaging and feature-tracking in a low-concentration sediment-laden flow. A phased array probe is used to generate a 2D ultrasound image at different times. Then, the feature points are extracted to be tracked instead of the centroids of the particle image. In order to better identify the corresponding feature point, each feature is described by an oriented angle and its location. Then, a statistical interpolation procedure is used to yield the displacement vector on the desired grid point. Finally a correction procedure is adopted because the ultrasound image is sequentially acquired line by line through the field of view. A simple test experiment was carried out to evaluate the performance. The ultrasound PTV system was applied to a sediment-laden flow with a low concentration of 1‰, and the speed was up to 10 cm s-1. In comparison to optical particle image velocimetry (PIV), ultrasound imaging does not have a limitation in optical access. The feature-tracking method does not have a binarisation and segmentation procedure, which can result in overlapping particles or a serious loss of particle data. The feature-tracking algorithm improves the peak locking effect and measurement accuracy. Thus, the ultrasound PTV algorithm is a feasible alternative and is significantly more robust against gradients than the correlation-based PIV algorithms in a low-concentration sediment-laden fluid.

  20. In Situ Synthesized Silver Nanoclusters for Tracking the Role of Telomerase Activity in the Differentiation of Mesenchymal Stem Cells to Neural Stem Cells.

    Dong, Fangyuan; Feng, Enduo; Zheng, Tingting; Tian, Yang

    2018-01-17

    Human mesenchymal stem cells (hMSCs) have potential use in cell replacement therapy for central nervous system disorders. However, the factors that impacted the differentiation process are unclear at the present stage because the powerful analytical method is the bottleneck. Herein, a novel strategy was developed for self-imaging and biosensing of telomerase activity in stem cells, using in situ biosynthesized silver nanoclusters (AgNCs) full of C bases. The present AgNCs possess synthetic convenience, long-time stability, and cytocompatibility. The weak fluorescence of these AgNCs is quickly turned on when approaching telomerase because of the strong interaction between C bases on AgNCs and G bases in telomerase, resulting in telomerase-dependent fluorescent signals. The developed method demonstrated high sensitivity and selectivity and broad dynamic linear range with a low detection limit. Using this powerful tool, it was first discovered that telomerase activity plays important roles in the proliferation of hMSCs and neural stem cells (NSCs) as well as during the differentiation processes from hMSCs to NSCs.

  1. ATLAS FTK Fast Track Trigger

    Iizawa, T; The ATLAS collaboration

    2014-01-01

    The Fast TracKer (FTK) will perform global track reconstruction after each Level-1 trigger accept signal to enable the software-based higher level trigger to have early access to tracking information. FTK is a dedicated processor based on a mixture of advanced technologies. Modern, powerful Field Programmable Gate Arrays (FPGAs) form an important part of the system architecture, and the large level of computing power required for pattern recognition is provided by incorporating standard-cell ASICs named Associative Memory (AM). Motivation and the architecture of the FTK system will be presented, and the status of hardware and simulation will be following.

  2. Report of the Central Tracking Group

    Cassel, D.G.; Hanson, G.G.

    1986-10-01

    Issues involved in building a realistic central tracking system for a general-purpose 4π detector for the SSC are addressed. Such a central tracking system must be capable of running at the full design luminosity of 10 33 cm -2 s -1 . Momentum measurement was required in a general-purpose 4π detector. Limitations on charged particle tracking detectors at the SSC imposed by rates and radiation damage are reviewed. Cell occupancy is the dominant constraint, which led us to the conclusion that only small cells, either wires or straw tubes, are suitable for a central tracking system at the SSC. Mechanical problems involved in building a central tracking system of either wires or straw tubes were studied, and our conclusion was that it is possible to build such a large central tracking system. Of course, a great deal of research and development is required. We also considered central tracking systems made of scintillating fibers or silicon microstrips, but our conclusion was that neither is a realistic candidate given the current state of technology. We began to work on computer simulation of a realistic central tracking system. Events from interesting physics processes at the SSC will be complex and will be further complicated by hits from out-of-time bunch crossings and multiple interactions within the same bunch crossing. Detailed computer simulations are needed to demonstrate that the pattern recognition and tracking problems can be solved

  3. CyberKnife with Tumor Tracking: An Effective Treatment for High-Risk Surgical Patients with Stage I Non-Small Cell Lung Cancer

    Chen, Viola J.; Oermann, Eric [Department of Radiation Medicine, Georgetown University Hospital, Washington, DC (United States); Vahdat, Saloomeh [Department of Pathology, Georgetown University Hospital, Washington, DC (United States); Rabin, Jennifer; Suy, Simeng; Yu, Xia; Collins, Sean P. [Department of Radiation Medicine, Georgetown University Hospital, Washington, DC (United States); Subramaniam, Deepa [Division of Hematology and Oncology, Georgetown University Hospital, Washington, DC (United States); Banovac, Filip [Department of Radiology, Georgetown University Hospital, Washington, DC (United States); Anderson, Eric [Division of Pulmonary, Critical Care and Sleep Medicine, Georgetown University Hospital, Washington, DC (United States); Collins, Brian T., E-mail: collinsb@gunet.georgetown.edu [Department of Radiation Medicine, Georgetown University Hospital, Washington, DC (United States)

    2012-02-01

    Published data suggests that wedge resection for stage I non-small cell lung cancer (NSCLC) is associated with improved overall survival compared to stereotactic body radiation therapy. We report CyberKnife outcomes for high-risk surgical patients with biopsy-proven stage I NSCLC. PET/CT imaging was completed for staging. Three-to-five gold fiducial markers were implanted in or near tumors to serve as targeting references. Gross tumor volumes (GTVs) were contoured using lung windows; the margins were expanded by 5 mm to establish the planning treatment volume (PTV). Treatment plans were designed using a mean of 156 pencil beams. Doses delivered to the PTV ranged from 42 to 60 Gy in three fractions. The 30 Gy isodose contour extended at least 1 cm from the GTV to eradicate microscopic disease. Treatments were delivered using the CyberKnife system with tumor tracking. Examination and PET/CT imaging occurred at 3 month follow-up intervals. Forty patients (median age 76) with a median maximum tumor diameter of 2.6 cm (range, 1.4–5.0 cm) and a mean post-bronchodilator percent predicted forced expiratory volume in 1 s (FEV1) of 57% (range, 21–111%) were treated. A median dose of 48 Gy was delivered to the PTV over 3–13 days (median, 7 days). The 30 Gy isodose contour extended a mean 1.9 cm from the GTV. At a median 44 months (range, 12–72 months) follow-up, the 3 year Kaplan–Meier locoregional control and overall survival estimates compare favorably with contemporary wedge resection outcomes at 91 and 75%, respectively. CyberKnife is an effective treatment approach for stage I NSCLC that is similar to wedge resection, eradicating tumors with 1–2 cm margins in order to preserve lung function. Prospective randomized trials comparing CyberKnife with wedge resection are necessary to confirm equivalence.

  4. Process and Information Tracking of Polycrystalline silicon Ingot for Solar Cell%铸锭多晶硅电池生产流程及信息跟踪

    焦富强; 乔卉莹

    2014-01-01

    Si-based photovoltaic materials account for a large proportion in the field of new energy, in which polycrystalline silicon ingot for solar cell is the main type. Many procedures must be used for production of the poly-crystalline silicon solar cell, therefore, accurate recording and tracking information of stuff and procedures play an im-portant role in technical improvement. In this paper, process and information tracking of every procedure in produc-tion of polycrystalline silicon solar cell are discussed, and easy encountered problems in information tracking are ana-lyzed.%在新能源开发利用领域硅基光伏材料占有较大比重,其中铸锭多晶硅光伏电池是当前太阳能电池的主要品种。生产多晶硅电池需要经历众多的加工工序,准确有序记录和跟踪物料流向及各工序相关信息是工艺研究和技术改进的基础。就铸锭多晶硅电池片生产流程及各工序信息跟踪问题进行了论述,并对实施信息跟踪时易出现的问题进行了分析。

  5. A method for three-dimensional interfacial particle image velocimetry (3D-IPIV) of an air–water interface

    Turney, Damon E; Anderer, Angelika; Banerjee, Sanjoy

    2009-01-01

    A new stereoscopic method for collecting particle image velocimetry (PIV) measurements within ∼1 mm of a wavy air–water interface with simultaneous measurements of the morphology of the interface is described. The method, termed three-dimensional interfacial particle image velocimetry (3D-IPIV), is tested in a wind wave channel with a wind speed of 5.8 m s −1 , water depth of 10 cm and a fetch of ∼9 m. Microscale breaking waves populate the interface and their flow patterns are clearly visible in the velocimetry results. The associated capillary waves and surface divergence patterns are observed. Several statistical measurements of the flow are compared with independent measurements and good agreement is found. The method shows great promise for investigating the transfer of momentum, heat and gases across an air–water interface, both in the laboratory and in field settings. Additional methods are described for manufacturing the flow tracers needed for the 3D-IPIV method. These tracers are likely to be useful for other researchers, and have the characteristics of being fluorescent, neutrally buoyant, non-toxic, monodisperse, inexpensive and easy to manufacture

  6. Computational fluid dynamics and particle image velocimetry assisted design tools for a new generation of trochoidal gear pumps

    M Garcia-Vilchez

    2015-06-01

    Full Text Available Trochoidal gear pumps produce significant flow pulsations that result in pressure pulsations, which interact with the system where they are connected, shortening the life of both the pump and circuit components. The complicated aspects of the operation of a gerotor pump make computational fluid dynamics the proper tool for modelling and simulating its flow characteristics. A three-dimensional model with deforming mesh computational fluid dynamics is presented, including the effects of the manufacturing tolerance and the leakage inside the pump. A new boundary condition is created for the simulation of the solid contact in the interteeth radial clearance. The experimental study of the pump is carried out by means of time-resolved particle image velocimetry, and results are qualitatively evaluated, thanks to the numerical simulation results. Time-resolved particle image velocimetry is developed in order to adapt it to the gerotor pump, and it is proved to be a feasible alternative to obtain the instantaneous flow of the pump in a direct mode, which would allow the determination of geometries that minimize the non-desired flow pulsations. Thus, a new methodology involving computational fluid dynamics and time-resolved particle image velocimetry is presented, which allows the obtaining of the instantaneous flow of the pump in a direct mode without altering its behaviour significantly.

  7. Track structure in biological models.

    Curtis, S B

    1986-01-01

    High-energy heavy ions in the galactic cosmic radiation (HZE particles) may pose a special risk during long term manned space flights outside the sheltering confines of the earth's geomagnetic field. These particles are highly ionizing, and they and their nuclear secondaries can penetrate many centimeters of body tissue. The three dimensional patterns of ionizations they create as they lose energy are referred to as their track structure. Several models of biological action on mammalian cells attempt to treat track structure or related quantities in their formulation. The methods by which they do this are reviewed. The proximity function is introduced in connection with the theory of Dual Radiation Action (DRA). The ion-gamma kill (IGK) model introduces the radial energy-density distribution, which is a smooth function characterizing both the magnitude and extension of a charged particle track. The lethal, potentially lethal (LPL) model introduces lambda, the mean distance between relevant ion clusters or biochemical species along the track. Since very localized energy depositions (within approximately 10 nm) are emphasized, the proximity function as defined in the DRA model is not of utility in characterizing track structure in the LPL formulation.

  8. Small-scale deflagration cylinder test with velocimetry wall-motion diagnostics

    Hooks, Daniel E [Los Alamos National Laboratory; Hill, Larry G [Los Alamos National Laboratory; Pierce, Timothy H [Los Alamos National Laboratory

    2010-01-01

    Predicting the likelihood and effects of outcomes resultant from thermal initiation of explosives remains a significant challenge. For certain explosive formulations, the general outcome can be broadly predicted given knowledge of certain conditions. However, there remain unexplained violent events, and increased statistical understanding of outcomes as a function of many variables, or 'violence categorization,' is needed. Additionally, the development of an equation of state equivalent for deflagration would be very useful in predicting possible detailed event consequences using traditional hydrodynamic detonation moders. For violence categorization, it is desirable that testing be efficient, such that it is possible to statistically define outcomes reliant on the processes of initiation of deflagration, steady state deflagration, and deflagration to detonation transitions. If the test simultaneously acquires information to inform models of violent deflagration events, overall predictive capabilities for event likelihood and consequence might improve remarkably. In this paper we describe an economical scaled deflagration cylinder test. The cyclotetramethylene tetranitramine (HMX) based explosive formu1lation PBX 9501 was tested using different temperature profiles in a thick-walled copper cylindrical confiner. This test is a scaled version of a recently demonstrated deflagration cylinder test, and is similar to several other thermal explosion tests. The primary difference is the passive velocimetry diagnostic, which enables measurement of confinement vessel wall velocities at failure, regardless of the timing and location of ignition.

  9. Local velocity measurements in lead-bismuth and sodium flows using the ultrasound doppler velocimetry

    Eckert, S.; Gerbeth, G.

    2003-01-01

    We will present measurements of the velocity profiles in liquid sodium and eutectic lead-bismuth by means of the Ultrasonic Doppler Velocimetry (UDV). A sodium flow in a rectangular duct exposed to an external, transverse magnetic field has been examined. To demonstrate the capability of UDV the transformation of the well-known turbulent, piston-like profile to an M-shaped velocity profile for growing magnetic field strength was observed. The significance of artifacts such as caused by the existence of reflecting interfaces in the measuring domain will be discussed. In the sodium case, the measurements were performed through the channel wall. An integrated ultrasonic sensor with acoustic wave-guide has been developed to overcome the limitation of ultrasonic transducers to temperatures lower than 200 .deg. C. This sensor can presently be applied at maximum temperatures up to 800 .deg. C. Stable and robust measurements have been performed in various PbBi flows in our laboratory at FZR as well as at the THESYS loop of the KALLA laboratory of the ForschungsZentrum Karlsruhe (FZK). We will also present experimental results obtained in a PbBi bubbly flow at 250...300 .deg. C. Argon bubbles were injected through a single orifice in a cylindrical container filled with stagnant PbBi. Velocity profiles were measured in the bubble plume. Mean values of the liquid as well as the bubble velocity were extracted from the data and will be presented as function of the gas flow rate

  10. Velocity profile measurement of lead-lithium flows by high-temperature ultrasonic doppler velocimetry

    Ueki, Y.; Kunugi, T.; Hirabayashi, Masaru; Nagai, Keiichi; Saito, Junichi; Ara, Kuniaki; Morley, N.B.

    2014-01-01

    This paper describes a high-temperature ultrasonic Doppler Velocimetry (HT-UDV) technique that has been successfully applied to measure velocity profiles of the lead-lithium eutectic alloy (PbLi) flows. The impact of tracer particles is investigated to determine requirements for HT-UDV measurement of PbLi flows. The HT-UDV system is tested on a PbLi flow driven by a rotating-disk in an inert atmosphere. We find that a sufficient amount of particles contained in the molten PbLi are required to successfully measure PbLi velocity profiles by HT-UDV. An X-ray diffraction analysis is performed to identify those particles in PbLi, and indicates that those particles were made of the lead mono-oxide (PbO). Since the specific densities of PbLi and PbO are close to each other, the PbO particles are expected to be well-dispersed in the bulk of molten PbLi. We conclude that the excellent dispersion of PbO particles enables in HT-UDV to obtain reliable velocity profiles for operation times of around 12 hours. (author)

  11. Particle image velocimetry measurements of the flow in the converging region of two parallel jets

    Wang, Huhu, E-mail: huhuwang@tamu.edu; Lee, Saya, E-mail: sayalee@tamu.edu; Hassan, Yassin A., E-mail: y-hassan@tamu.edu

    2016-09-15

    Highlights: • The flow behaviors in the converging region were non-intrusively investigated using PIV. • The PIV results using two measuring scales and LDV data matched very well. • Significant momentum transfer was observed in the merging region right after the merging point. • Instantaneous vector field revealed characteristic interacting patterns of the jets. - Abstract: The interaction between parallel jets plays a critical role in determining the characteristics of the momentum and heat transfer in the flow. Specifically for next generation VHTR, the output temperature will be about 900 °C, and any thermal oscillations will create safety issues. The mixing variations of the coolants in the reactor core may influence these power oscillations. Numerous numerical tools such as computational fluid dynamics (CFD) simulations have been used to support the reactor design. The validation of CFD method is important to ensure the fidelity of the calculations. This requires high-fidelity, qualified benchmark data. Particle image velocimetry (PIV), a non-intrusive measuring technique, was used to provide benchmark data for resolving a simultaneous flow field in the converging region of two submerged parallel jets issued from rectangular channels. The jets studied in this work had an equal discharge velocity at room temperature. The turbulent characteristics including the distributions of mean velocities, turbulence intensities, Reynolds stresses and z-component vorticity were studied. The streamwise mean velocity measured by PIV and LDV were compared, and they agreed very well.

  12. Ultrasonic particle image velocimetry for improved flow gradient imaging: algorithms, methodology and validation

    Niu Lili; Qian Ming; Yu Wentao; Jin Qiaofeng; Ling Tao; Zheng Hairong; Wan Kun; Gao Shen

    2010-01-01

    This paper presents a new algorithm for ultrasonic particle image velocimetry (Echo PIV) for improving the flow velocity measurement accuracy and efficiency in regions with high velocity gradients. The conventional Echo PIV algorithm has been modified by incorporating a multiple iterative algorithm, sub-pixel method, filter and interpolation method, and spurious vector elimination algorithm. The new algorithms' performance is assessed by analyzing simulated images with known displacements, and ultrasonic B-mode images of in vitro laminar pipe flow, rotational flow and in vivo rat carotid arterial flow. Results of the simulated images show that the new algorithm produces much smaller bias from the known displacements. For laminar flow, the new algorithm results in 1.1% deviation from the analytically derived value, and 8.8% for the conventional algorithm. The vector quality evaluation for the rotational flow imaging shows that the new algorithm produces better velocity vectors. For in vivo rat carotid arterial flow imaging, the results from the new algorithm deviate 6.6% from the Doppler-measured peak velocities averagely compared to 15% of that from the conventional algorithm. The new Echo PIV algorithm is able to effectively improve the measurement accuracy in imaging flow fields with high velocity gradients.

  13. Visualization of nasal airflow patterns in a patient affected with atrophic rhinitis using particle image velocimetry

    Garcia, G J M [Hamner Institutes for Health Sciences, NC (United States); Mitchell, G [The Queens University of Belfast, Belfast (United Kingdom); Bailie, N [The Queens University of Belfast, Belfast (United Kingdom); Thornhill, D [The Queens University of Belfast, Belfast (United Kingdom); Watterson, J [The Queens University of Belfast, Belfast (United Kingdom); Kimbell, J S [Hamner Institutes for Health Sciences, NC (United States)

    2007-10-15

    The relationship between airflow patterns in the nasal cavity and nasal function is poorly understood. This paper reports an experimental study of the interplay between symptoms and airflow patterns in a patient affected with atrophic rhinitis. This pathology is characterized by mucosal dryness, fetor, progressive atrophy of anatomical structures, a spacious nasal cavity, and a paradoxical sensation of nasal congestion. A physical replica of the patient's nasal geometry was made and particle image velocimetry (PIV) was used to visualize and measure the flow field. The nasal replica was based on computed tomography (CT) scans of the patient and was built in three steps: three-dimensional reconstruction of the CT scans; rapid prototyping of a cast; and sacrificial use of the cast to form a model of the nasal passage in clear silicone. Flow patterns were measured by running a water-glycerol mixture through the replica and evaluating the displacement of particles dispersed in the liquid using PIV. The water-glycerol flow rate used corresponded to an air flow rate representative of a human breathing at rest. The trajectory of the flow observed in the left passage of the nose (more affected by atrophic rhinitis) differed markedly from what is considered normal, and was consistent with patterns of epithelial damage observed in cases of the condition. The data are also useful for validation of computational fluid dynamics predictions.

  14. Study of particles clouds ejected under shock: the contributions of Photonic Doppler Velocimetry

    Prudhomme, Gabriel

    2014-01-01

    A metal plate subjected to a shock (tin, 10 GPa) undergoes a variety of damages such as spalling or the ejection of a cloud of particles. Two main mechanisms govern the formation of this cloud: the micro-jetting and the melting under shock. Photonic Doppler Velocimetry (PDV, a.k.a. LDV or het-V) is a multi-velocity time-resolved diagnostic. Developed from 2000's, the all-fibered conception makes its integration easy into shock experiments. The purpose of the thesis is to describe the contributions of PDV systems for high-velocity (several km/s) particle-cloud characterization, including micro-jetting cloud. This document presents a state of the art of shock generators, diagnostics and (numerical and experimental) studies involved in metallic micro-machined jetting. An extensive study of a PDV system is proposed. It leads to the definition of time-velocity spectrogram, evaluated in units of collected power, and a detection capability limit. Thanks to photon diffusion models, a threshold in the diameter of the measured particle is estimated. A PDV spectrogram simulation program is shown within the framework of particle clouds. Finally, several experimental campaigns are exposed. They emphasize the remarkable capacities of the system; results are compared to simulations. Diameter distributions are inferred using slowing down in air or in other gazes. Some radiometric analyses are also performed. (author) [fr

  15. Comparison of Global Sizing Velocimetry and Phase Doppler Anemometry measurements of alternative jet fuel sprays

    Sadr, Reza; Kannaiyan, Kumaran

    2013-11-01

    Atomization plays a crucial precursor role in liquid fuel combustion that directly affects the evaporation, mixing, and emission levels. Laser diagnostic techniques are often used to study the spray characteristics of liquid fuels. The objective of this work is to compare the spray measurements of Gas-to Liquid (GTL) jet fuels obtained using Global Sizing Velocimetry (GSV) and Phase Doppler Anemometry (PDA) techniques at global and local levels, respectively. The chemical and physical properties of GTL fuels are different from conventional jet fuels, owing to the difference in their production methodology. In this work, the experimental facility, the measurement techniques, and spray characteristics of two different GTL fuels are discussed and compared with those of Jet A-1 fuel. Results clearly demonstrate that although the global measurement gives an overall picture of the spray, fine details are obtained only through local measurements and complement in gaining more inferences into the spray characteristics. The results also show a close similarity in spray characteristics between GTL and Jet A-1 fuels. Funded by Qatar Science and Technology Park.

  16. Velocity Deficits in the Wake of Model Lemon Shark Dorsal Fins Measured with Particle Image Velocimetry

    Terry, K. N.; Turner, V.; Hackett, E.

    2017-12-01

    Aquatic animals' morphology provides inspiration for human technological developments, as their bodies have evolved and become adapted for efficient swimming. Lemon sharks exhibit a uniquely large second dorsal fin that is nearly the same size as the first fin, the hydrodynamic role of which is unknown. This experimental study looks at the drag forces on a scale model of the Lemon shark's unique two-fin configuration in comparison to drag forces on a more typical one-fin configuration. The experiments were performed in a recirculating water flume, where the wakes behind the scale models are measured using particle image velocimetry. The experiments are performed at three different flow speeds for both fin configurations. The measured instantaneous 2D distributions of the streamwise and wall-normal velocity components are ensemble averaged to generate streamwise velocity vertical profiles. In addition, velocity deficit profiles are computed from the difference between these mean streamwise velocity profiles and the free stream velocity, which is computed based on measured flow rates during the experiments. Results show that the mean velocities behind the fin and near the fin tip are smallest and increase as the streamwise distance from the fin tip increases. The magnitude of velocity deficits increases with increasing flow speed for both fin configurations, but at all flow speeds, the two-fin configurations generate larger velocity deficits than the one-fin configurations. Because the velocity deficit is directly proportional to the drag force, these results suggest that the two-fin configuration produces more drag.

  17. Flow mapping of multiphase flows using a novel single stem endoscopic particle image velocimetry instrument

    Lad, N; Adebayo, D; Aroussi, A

    2011-01-01

    Particle image velocimetry (PIV) is a successful flow mapping technique which can optically quantify large portions of a flow regime. This enables the method to be completely non-intrusive. The ability to be non-intrusive to any flow has allowed PIV to be used in a large range of industrial sectors for many applications. However, a fundamental disadvantage of the conventional PIV technique is that it cannot easily be used with flows which have no or limited optical access. Flows which have limited optical access for PIV measurement have been addressed using endoscopic PIV techniques. This system uses two separate probes which relay a light sheet and imaging optics to a planar position within the desired flow regime. This system is effective in medical and engineering applications. The present study has been involved in the development of a new endoscopic PIV system which integrates the illumination and imaging optics into one rigid probe. This paper focuses on the validation of the images taken from the novel single stem endoscopic PIV system. The probe is used within atomized spray flow and is compared with conventional PIV measurement and also pitot-static data. The endoscopic PIV system provides images which create localized velocity maps that are comparable with the global measurement of the conventional PIV system. The velocity information for both systems clearly show similar results for the spray characterization and are also validated using the pitot-static data

  18. A 3D velocimetry study of the flow through prosthetic heart valves

    Ledesma, R.; Zenit, R.; Pulos, G.; Sanchez, E.; Juarez, A.

    2006-11-01

    Blood damage commonly appears in medical valve prothesis. It is a mayor concern for the designers and surgeons. It is well known that this damage and other complications result from the modified fluid dynamics through the replacement valve. To evaluate the performance of prosthetic heart valves, it is necessary to study the flow through them. To conduct this study , we have built a flow channel that emulates cardiac conditions and allows optical access such that a 3D-PIV velocimetry system could be used. The experiments are aimed to reconstruct the downstream structure of the flow through a mechanical and a bio-material tricuspid heart valve prothesis. Preliminary results show that the observed coherent structures can be related with haemolysis and trombosis, illnesses commonly found in valve prothesis recipients. The mean flow, the levels of strain rate and the turbulence intensity generated by the valves can also be directly related to blood damage. In general, bio-material made valves tend to reduce these complications.

  19. Experimental characterization of solid particle transport by slug flow using Particle Image Velocimetry

    Goharzadeh, A; Rodgers, P

    2009-01-01

    This paper presents an experimental study of gas-liquid slug flow on solid particle transport inside a horizontal pipe with two types of experiments conducted. The influence of slug length on solid particle transportation is characterized using high speed photography. Using combined Particle Image Velocimetry (PIV) with Refractive Index Matching (RIM) and fluorescent tracers (two-phase oil-air loop) the velocity distribution inside the slug body is measured. Combining these experimental analyses, an insight is provided into the physical mechanism of solid particle transportation due to slug flow. It was observed that the slug body significantly influences solid particle mobility. The physical mechanism of solid particle transportation was found to be discontinuous. The inactive region (in terms of solid particle transport) upstream of the slug nose was quantified as a function of gas-liquid composition and solid particle size. Measured velocity distributions showed a significant drop in velocity magnitude immediately upstream of the slug nose and therefore the critical velocity for solid particle lifting is reached further upstream.

  20. Ghost Particle Velocimetry implementation in millimeters devices and comparison with μPIV

    Riccomi, Marco; Alberini, Federico; Brunazzi, Elisabetta; Vigolo, Daniele

    2016-11-01

    Micro/milli-fluidic devices are becoming an important reference for several disciplines and are quickly increasing their applications in scientific, as well as industrial, environment. As a consequence, the development of techniques able to analyse these kinds of systems is required to allow their progress. Here we show the implementation of the Ghost Particle Velocimetry (GPV) for the flow velocity field investigation in milli-fluidic devices. This innovative technique has been recently introduced, and has been already proven to be useful in describing rapid phenomenon at a small scale. In this work, the GPV has been used to characterize the trapping of light suspended material in a branching junction. Experiments have been performed to identify the flow velocity field close to a millimeters scale T-junction, at different Reynolds numbers. Particularly interesting are the complex structures, such as vortices and recirculation zones, induced by the vortex breakdown phenomenon. The results obtained have been deeply validated and compared with the well-established μPIV, highlighting the differences in terms of qualitative and quantitative parameters. A performance comparison has been designed to underline the strengths and weaknesses of the two experimental techniques.