WorldWideScience

Sample records for two-photon time-lapse imaging

  1. Time-lapse Raman imaging of osteoblast differentiation

    Science.gov (United States)

    Hashimoto, Aya; Yamaguchi, Yoshinori; Chiu, Liang-Da; Morimoto, Chiaki; Fujita, Katsumasa; Takedachi, Masahide; Kawata, Satoshi; Murakami, Shinya; Tamiya, Eiichi

    2015-07-01

    Osteoblastic mineralization occurs during the early stages of bone formation. During this mineralization, hydroxyapatite (HA), a major component of bone, is synthesized, generating hard tissue. Many of the mechanisms driving biomineralization remain unclear because the traditional biochemical assays used to investigate them are destructive techniques incompatible with viable cells. To determine the temporal changes in mineralization-related biomolecules at mineralization spots, we performed time-lapse Raman imaging of mouse osteoblasts at a subcellular resolution throughout the mineralization process. Raman imaging enabled us to analyze the dynamics of the related biomolecules at mineralization spots throughout the entire process of mineralization. Here, we stimulated KUSA-A1 cells to differentiate into osteoblasts and conducted time-lapse Raman imaging on them every 4 hours for 24 hours, beginning 5 days after the stimulation. The HA and cytochrome c Raman bands were used as markers for osteoblastic mineralization and apoptosis. From the Raman images successfully acquired throughout the mineralization process, we found that β-carotene acts as a biomarker that indicates the initiation of osteoblastic mineralization. A fluctuation of cytochrome c concentration, which indicates cell apoptosis, was also observed during mineralization. We expect time-lapse Raman imaging to help us to further elucidate osteoblastic mineralization mechanisms that have previously been unobservable.

  2. TimeLapseAnalyzer: Multi-target analysis for live-cell imaging and time-lapse microscopy

    DEFF Research Database (Denmark)

    Huth, Johannes; Buchholz, Malte; Kraus, Johann M.

    2011-01-01

    The direct observation of cells over time using time-lapse microscopy can provide deep insights into many important biological processes. Reliable analyses of motility, proliferation, invasive potential or mortality of cells are essential to many studies involving live cell imaging and can aid in...... counting and tube formation analysis in high throughput screening of live-cell experiments. TimeLapseAnalyzer is freely available (MATLAB, Open Source) at http://www.informatik.uniulm. de/ni/mitarbeiter/HKestler/tla......., we developed TimeLapseAnalyzer. Apart from general purpose image enhancements and segmentation procedures, this extensible, self-contained, modular cross-platform package provides dedicated modalities for fast and reliable analysis of multi-target cell tracking, scratch wound healing analysis, cell...

  3. Two-photon time-lapse microscopy of BODIPY-cholesterol reveals anomalous sterol diffusion in chinese hamster ovary cells

    Directory of Open Access Journals (Sweden)

    Lund Frederik W

    2012-10-01

    Full Text Available Abstract Background Cholesterol is an important membrane component, but our knowledge about its transport in cells is sparse. Previous imaging studies using dehydroergosterol (DHE, an intrinsically fluorescent sterol from yeast, have established that vesicular and non-vesicular transport modes contribute to sterol trafficking from the plasma membrane. Significant photobleaching, however, limits the possibilities for in-depth analysis of sterol dynamics using DHE. Co-trafficking studies with DHE and the recently introduced fluorescent cholesterol analog BODIPY-cholesterol (BChol suggested that the latter probe has utility for prolonged live-cell imaging of sterol transport. Results We found that BChol is very photostable under two-photon (2P-excitation allowing the acquisition of several hundred frames without significant photobleaching. Therefore, long-term tracking and diffusion measurements are possible. Two-photon temporal image correlation spectroscopy (2P-TICS provided evidence for spatially heterogeneous diffusion constants of BChol varying over two orders of magnitude from the cell interior towards the plasma membrane, where D ~ 1.3 μm2/s. Number and brightness (N&B analysis together with stochastic simulations suggest that transient partitioning of BChol into convoluted membranes slows local sterol diffusion. We observed sterol endocytosis as well as fusion and fission of sterol-containing endocytic vesicles. The mobility of endocytic vesicles, as studied by particle tracking, is well described by a model for anomalous subdiffusion on short time scales with an anomalous exponent α ~ 0.63 and an anomalous diffusion constant of Dα = 1.95 x 10-3 μm2/sα. On a longer time scale (t > ~5 s, a transition to superdiffusion consistent with slow directed transport with an average velocity of v ~ 6 x 10-3 μm/s was observed. We present an analytical model that bridges the two regimes and fit this model to vesicle

  4. Time-lapse imaging of neural development: zebrafish lead the way into the fourth dimension.

    Science.gov (United States)

    Rieger, Sandra; Wang, Fang; Sagasti, Alvaro

    2011-07-01

    Time-lapse imaging is often the only way to appreciate fully the many dynamic cell movements critical to neural development. Zebrafish possess many advantages that make them the best vertebrate model organism for live imaging of dynamic development events. This review will discuss technical considerations of time-lapse imaging experiments in zebrafish, describe selected examples of imaging studies in zebrafish that revealed new features or principles of neural development, and consider the promise and challenges of future time-lapse studies of neural development in zebrafish embryos and adults. Copyright © 2011 Wiley-Liss, Inc.

  5. Automatic segmentation of time-lapse microscopy images depicting a live Dharma embryo.

    Science.gov (United States)

    Zacharia, Eleni; Bondesson, Maria; Riu, Anne; Ducharme, Nicole A; Gustafsson, Jan-Åke; Kakadiaris, Ioannis A

    2011-01-01

    Biological inferences about the toxicity of chemicals reached during experiments on the zebrafish Dharma embryo can be greatly affected by the analysis of the time-lapse microscopy images depicting the embryo. Among the stages of image analysis, automatic and accurate segmentation of the Dharma embryo is the most crucial and challenging. In this paper, an accurate and automatic segmentation approach for the segmentation of the Dharma embryo data obtained by fluorescent time-lapse microscopy is proposed. Experiments performed in four stacks of 3D images over time have shown promising results.

  6. Advances in interpretation of subsurface processes with time-lapse electrical imaging

    Science.gov (United States)

    Singha, Kaminit; Day-Lewis, Frederick D.; Johnson, Tim B.; Slater, Lee D.

    2015-01-01

    Electrical geophysical methods, including electrical resistivity, time-domain induced polarization, and complex resistivity, have become commonly used to image the near subsurface. Here, we outline their utility for time-lapse imaging of hydrological, geochemical, and biogeochemical processes, focusing on new instrumentation, processing, and analysis techniques specific to monitoring. We review data collection procedures, parameters measured, and petrophysical relationships and then outline the state of the science with respect to inversion methodologies, including coupled inversion. We conclude by highlighting recent research focused on innovative applications of time-lapse imaging in hydrology, biology, ecology, and geochemistry, among other areas of interest.

  7. CellProfiler Tracer: exploring and validating high-throughput, time-lapse microscopy image data.

    Science.gov (United States)

    Bray, Mark-Anthony; Carpenter, Anne E

    2015-11-04

    Time-lapse analysis of cellular images is an important and growing need in biology. Algorithms for cell tracking are widely available; what researchers have been missing is a single open-source software package to visualize standard tracking output (from software like CellProfiler) in a way that allows convenient assessment of track quality, especially for researchers tuning tracking parameters for high-content time-lapse experiments. This makes quality assessment and algorithm adjustment a substantial challenge, particularly when dealing with hundreds of time-lapse movies collected in a high-throughput manner. We present CellProfiler Tracer, a free and open-source tool that complements the object tracking functionality of the CellProfiler biological image analysis package. Tracer allows multi-parametric morphological data to be visualized on object tracks, providing visualizations that have already been validated within the scientific community for time-lapse experiments, and combining them with simple graph-based measures for highlighting possible tracking artifacts. CellProfiler Tracer is a useful, free tool for inspection and quality control of object tracking data, available from http://www.cellprofiler.org/tracer/.

  8. Seismic time-lapse imaging using Interferometric least-squares migration

    KAUST Repository

    Sinha, Mrinal

    2016-09-06

    One of the problems with 4D surveys is that the environmental conditions change over time so that the experiment is insufficiently repeatable. To mitigate this problem, we propose the use of interferometric least-squares migration (ILSM) to estimate the migration image for the baseline and monitor surveys. Here, a known reflector is used as the reference reflector for ILSM. Results with synthetic and field data show that ILSM can eliminate artifacts caused by non-repeatability in time-lapse surveys.

  9. Seismic time-lapse imaging using Interferometric least-squares migration

    KAUST Repository

    Sinha, Mrinal; Schuster, Gerard T.

    2016-01-01

    One of the problems with 4D surveys is that the environmental conditions change over time so that the experiment is insufficiently repeatable. To mitigate this problem, we propose the use of interferometric least-squares migration (ILSM) to estimate the migration image for the baseline and monitor surveys. Here, a known reflector is used as the reference reflector for ILSM. Results with synthetic and field data show that ILSM can eliminate artifacts caused by non-repeatability in time-lapse surveys.

  10. Imaging the developing heart: synchronized time-lapse microscopy during developmental changes

    Science.gov (United States)

    Nelson, Carl J.; Buckley, Charlotte; Mullins, John J.; Denvir, Martin A.; Taylor, Jonathan

    2018-02-01

    How do you use imaging to analyse the development of the heart, which not only changes shape but also undergoes constant, high-speed, quasi-periodic changes? We have integrated ideas from prospective and retrospective optical gating to capture long-term, phase-locked developmental time-lapse videos. In this paper we demonstrate the success of this approach over a key developmental time period: heart looping, where large changes in heart shape prevent previous prospective gating approaches from capturing phase- locked videos. We use the comparison with other approaches to in vivo heart imaging to highlight the importance of collecting the most appropriate data for the biological question.

  11. Time-lapse 3D imaging of calcite precipitation in a microporous column

    Science.gov (United States)

    Godinho, Jose R. A.; Withers, Philip J.

    2018-02-01

    Time-lapse X-ray computed tomography is used to image the evolution of calcite precipitation during flow through microporous quartz over the course of 400 h. The growth rate decreases by more than seven times, which is linked to the clogging of flow paths that restricts flow to some regions of the column. Fewer precipitates are observed as a function of column depth, which is found to be related to a differential nucleation density along the sample. A higher nucleation density closer to the inlet implies more crystal volume increase per unit of time without affecting the rate if normalized to the surface area of crystals. Our overall growth rates measured in porous media are orders of magnitude slower than growth rates derived from traditional precipitation experiments on free surfaces. Based on our time-lapse results we hypothesize a scenario where the evolving distribution of precipitates within a pore structure during precipitation progressively modifies the local transport through the pores. Within less permeable regions the saturation index may be lower than along the main flow paths. Therefore, the reactive crystal surfaces within those regions grow at a slower rate than that expected from the bulk fluid composition. Since the amount of reactive surface area within these less permeable regions increases over time, the overall growth rate decreases without a necessary significant change of the bulk fluid composition along more permeable flow paths. In conclusion, the overall growth rates in an evolving porous media expected from bulk fluid compositions alone can be overestimated due to the development of stagnant sub-regions where the reactive surface area is bath by a solution with lower saturation index. In this context we highlight the value of time-lapse 3D studies for understanding the dynamics of mineral precipitation in porous media.

  12. Time lapse imaging: is it time to incorporate this technology into routine clinical practice?

    Science.gov (United States)

    Bhide, Priya; Maheshwari, Abha; Cutting, Rachel; Seenan, Susan; Patel, Anita; Khan, Khalid; Homburg, Roy

    2017-06-01

    Time-lapse imaging (TLI) systems for embryo incubation, assessment and selection are a novel technology available to in vitro fertilization (IVF) clinics. However, there is uncertainty about their clinical and cost-effectiveness and insufficient good quality evidence to warrant their routine use. Despite this, enthusiastic commercial marketing and slipping clinical equipoise have led to the widespread hasty introduction of this technology into practice, often at a considerable expense to the patient. We have reviewed the published literature and aim to summarize the strengths, weaknesses, opportunities and threats of these systems. These specialized incubators provide undisturbed embryo culture conditions and, by almost continuous monitoring of embryo development, generate morphokinetic parameters to aid embryo selection. They are thus hypothesized to improve outcomes following IVF. Although literature reports improved reproductive outcomes, these outcomes are largely surrogate and there is a paucity of studies reporting live births. The use of time lapse systems may reduce early pregnancy loss, increase elective single embryo transfers and limit multiple pregnancies through better embryo selection. However, the quality of the studies and hence the evidence so far, is low to moderate quality. We recommend further research producing robust high-quality evidence for and against the use of these systems.

  13. Time-Lapse Electrical Resistivity Investigations for Imaging the Grouting Injection in Shallow Subsurface Cavities

    Directory of Open Access Journals (Sweden)

    Muhammad Farooq

    2014-01-01

    Full Text Available The highway of Yongweol-ri, Muan-gun, south-western part of the South Korean Peninsula, is underlain by the abandoned of subsurface cavities, which were discovered in 2005. These cavities lie at shallow depths with the range of 5∼15 meters below the ground surface. Numerous subsidence events have repeatedly occurred in the past few years, damaging infrastructure and highway. As a result of continuing subsidence issues, the Korean Institute of Geosciences and Mineral Resources (KIGAM was requested by local administration to resolve the issue. The KIGAM used geophysical methods to delineate subsurface cavities and improve more refined understanding of the cavities network in the study area. Cement based grouting has been widely employed in the construction industry to reinforce subsurface ground. In this research work, time-lapse electrical resistivity surveys were accomplished to monitor the grouting injection in the subsurface cavities beneath the highway, which have provided a quasi-real-time monitoring for modifying the subsurface cavities related to ground reinforcement, which would be difficult with direct methods. The results obtained from time-lapse electrical resistivity technique have satisfactory imaged the grouting injection experiment in the subsurface cavities beneath the highway. Furthermore, the borehole camera confirmed the presence of grouting material in the subsurface cavities, and hence this procedure increases the mechanical resistance of subsurface cavities below the highway.

  14. A poly(dimethylsiloxane)-based device enabling time-lapse imaging with high spatial resolution

    International Nuclear Information System (INIS)

    Hirano, Masahiko; Hoshida, Tetsushi; Sakaue-Sawano, Asako; Miyawaki, Atsushi

    2010-01-01

    We have developed a regulator-free device that enables long-term incubation of mammalian cells for epi-fluorescence imaging, based on a concept that the size of sample to be gassed and heated is reduced to observation scale. A poly(dimethylsiloxane) block stamped on a coverslip works as a long-lasting supplier of CO 2 -rich gas to adjust bicarbonate-containing medium in a tiny chamber at physiological pH, and an oil-immersion objective warms cells across the coverslip. A time-lapse imaging experiment using HeLa cells stably expressing fluorescent cell-cycle indicators showed that the cells in the chamber proliferated with normal cell-cycle period over 2 days.

  15. Microsphere imaging with confocal microscopy and two photon microscopy

    International Nuclear Information System (INIS)

    Chun, Hyung Su; An, Kyung Won; Lee, Jai Hyung

    2002-01-01

    We have acquired images of polystyrene and fused-silica microsphere by using conventional optical microscopy, confocal microscopy and two-photon microscopy, and performed comparative analysis of these images. Different from conventional optical microscopy, confocal and two-photon microscopy had good optical sectioning capability. In addition, confocal microscopy and two-photon microscopy had better lateral resolution than conventional optical microscopy. These results are attributed to confocality and nonlinearity of confocal microscopy and two photon microscopy, respectively.

  16. Non-rigid estimation of cell motion in calcium time-lapse images

    Science.gov (United States)

    Hachi, Siham; Lucumi Moreno, Edinson; Desmet, An-Sofie; Vanden Berghe, Pieter; Fleming, Ronan M. T.

    2016-03-01

    Calcium imaging is a widely used technique in neuroscience permitting the simultaneous monitoring of electro- physiological activity of hundreds of neurons at single cell resolution. Identification of neuronal activity requires rapid and reliable image analysis techniques, especially when neurons fire and move simultaneously over time. Traditionally, image segmentation is performed to extract individual neurons in the first frame of a calcium sequence. Thereafter, the mean intensity is calculated from the same region of interest in each frame to infer calcium signals. However, when cells move, deform and fire, this segmentation on its own generates artefacts and therefore biased neuronal activity. Therefore, there is a pressing need to develop a more efficient cell tracking technique. We hereby present a novel vision-based cell tracking scheme using a thin-plate spline deformable model. The thin-plate spline warping is based on control points detected using the Fast from Accelerated Segment Test descriptor and tracked using the Lucas-Kanade optical flow. Our method is able to track neurons in calcium time-series, even when there are large changes in intensity, such as during a firing event. The robustness and efficiency of the proposed approach is validated on real calcium time-lapse images of a neuronal population.

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

    Science.gov (United States)

    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.

  18. Metamorphosis revealed: time-lapse three-dimensional imaging inside a living chrysalis.

    Science.gov (United States)

    Lowe, Tristan; Garwood, Russell J; Simonsen, Thomas J; Bradley, Robert S; Withers, Philip J

    2013-07-06

    Studies of model insects have greatly increased our understanding of animal development. Yet, they are limited in scope to this small pool of model species: a small number of representatives for a hyperdiverse group with highly varied developmental processes. One factor behind this narrow scope is the challenging nature of traditional methods of study, such as histology and dissection, which can preclude quantitative analysis and do not allow the development of a single individual to be followed. Here, we use high-resolution X-ray computed tomography (CT) to overcome these issues, and three-dimensionally image numerous lepidopteran pupae throughout their development. The resulting models are presented in the electronic supplementary material, as are figures and videos, documenting a single individual throughout development. They provide new insight and details of lepidopteran metamorphosis, and allow the measurement of tracheal and gut volume. Furthermore, this study demonstrates early and rapid development of the tracheae, which become visible in scans just 12 h after pupation. This suggests that there is less remodelling of the tracheal system than previously expected, and is methodologically important because the tracheal system is an often-understudied character system in development. In the future, this form of time-lapse CT-scanning could allow faster and more detailed developmental studies on a wider range of taxa than is presently possible.

  19. Applications of quantitative time lapse holographic imaging to the development of complex pharmaceutical nano formulations

    Science.gov (United States)

    Luther, Ed; Mendes, Livia; Pan, Jiayi; Costa, Daniel; Sarisozen, Can; Torchilin, Vladimir

    2018-02-01

    We rely on in vitro cellular cultures to evaluate the effects of the components of multifunctional nano-based formulations under development. We employ an incubator-adapted, label-free holographic imaging cytometer HoloMonitor M4® (Phase Holographic Imaging, Lund, Sweden) to obtain multi-day time-lapse sequences at 5- minute intervals. An automated stage allows hand-free acquisition of multiple fields of view. Our system is based on the Mach-Zehnder interferometry principle to create interference patterns which are deconvolved to produce images of the optical thickness of the field of view. These images are automatically segmented resulting in a full complement of quantitative morphological features, such as optical volume, thickness, and area amongst many others. Precise XY cell locations and the time of acquisition are also recorded. Visualization is best achieved by novel 4-Dimensional plots, where XY position is plotted overtime time (Z-directions) and cell-thickness is coded as color or gray scale brightness. Fundamental events of interest, i.e., cells undergoing mitosis or mitotic dysfunction, cell death, cell-to-cell interactions, motility are discernable. We use both 2D and 3D models of the tumor microenvironment. We report our new analysis method to track feature changes over time based on a 4-sample version of the Kolmogorov-Smirnov test. Feature A is compared to Control A, and Feature B is compared to Control B to give a 2D probability plot of the feature changes over time. As a result, we efficiently obtain vectors quantifying feature changes over time in various sample conditions, i.e., changing compound concentrations or multi-compound combinations.

  20. Tracking snowmelt in the subsurface: time-lapse electrical resistivity imaging on an alpine hill slope.

    Science.gov (United States)

    Thayer, D.; Parsekian, A.; Hyde, K.; Beverly, D.; Speckman, H. N.; Ewers, B. E.

    2015-12-01

    In the mountain West region the winter snowpack provides more than 70% of our annual water supply. Modeling and predicting the timing and magnitude of snowmelt-driven water yield is difficult due to the complexities of hydrologic systems that move meltwater from snow to rivers. Particular challenges are understanding the temporal and spatial domain of subsurface hydraulic processes at relevant scales, which range from points to catchments. Subsurface characterization often requires borehole instrumentation, which is expensive and extremely difficult to install in remote, rugged terrain. Advancements in non-invasive geophysical methods allow us to monitor changes in geophysical parameters over time and infer changes in hydraulic processes. In the No-Name experimental catchment in the Medicine Bow National Forest in Wyoming, we are conducting a multi-season, time-lapse electrical resistivity imaging survey on a sub-alpine hill slope. This south-facing, partially forested slope ranges from 5 degrees to 35 degrees in steepness and consists of a soil mantle covering buried glacial talus deposits of unknown depth. A permanent grid of down-slope and cross-slope electrode arrays is monitored up to four times a day. The arrays span the entire vertical distance of the slope, from an exposed bedrock ridge to a seasonal drainage below, and cover treed and non-treed areas. Geophysical measurements are augmented by temperature and moisture time-series instrumented below the surface in a contiguous 3 meter borehole. A time-series of multiple resistivity models each day from May to July shows the changing distribution of subsurface moisture during a seasonal drying sequence punctuated by isolated rain events. Spatial patterns of changing moisture indicate that soil and gravel in the top two meters drain into a saturated layer parallel to the slope which overlies less saturated material. These results suggest that water from snowmelt and rain events tends to move down-slope beneath

  1. Segmentation Method of Time-Lapse Microscopy Images with the Focus on Biocompatibility Assessment

    Czech Academy of Sciences Publication Activity Database

    Soukup, Jindřich; Císař, P.; Šroubek, Filip

    2016-01-01

    Roč. 22, č. 3 (2016), s. 497-506 ISSN 1431-9276 R&D Projects: GA ČR GA13-29225S Grant - others:GA MŠk(CZ) LO1205; GA UK(CZ) 914813/2013; GA UK(CZ) SVV-2016-260332; CENAKVA(CZ) CZ.1.05/2.1.00/01.0024 Institutional support: RVO:67985556 Keywords : phase contrast microscopy * segmentation * biocompatibility assessment * time-lapse * cytotoxicity testing Subject RIV: JD - Computer Applications, Robotics Impact factor: 1.891, year: 2016 http://library.utia.cas.cz/separaty/2016/ZOI/soukupj-0460642.pdf

  2. A bioaccumulative cyclometalated platinum(II) complex with two-photon-induced emission for live cell imaging.

    Science.gov (United States)

    Koo, Chi-Kin; Wong, Ka-Leung; Man, Cornelia Wing-Yin; Lam, Yun-Wah; So, Leo King-Yan; Tam, Hoi-Lam; Tsao, Sai-Wah; Cheah, Kok-Wai; Lau, Kai-Chung; Yang, Yang-Yi; Chen, Jin-Can; Lam, Michael Hon-Wah

    2009-02-02

    The cyclometalated platinum(II) complex [Pt(L)Cl], where HL is a new cyclometalating ligand 2-phenyl-6-(1H-pyrazol-3-yl)pyridine containing C(phenyl), N(pyridyl), and N(pyrazolyl) donor moieties, was found to possess two-photon-induced luminescent properties. The two-photon-absorption cross section of the complex in N,N-dimethylformamide at room temperature was measured to be 20.8 GM. Upon two-photon excitation at 730 nm from a Ti:sapphire laser, bright-green emission was observed. Besides its two-photon-induced luminescent properties, [Pt(L)Cl] was able to be rapidly accumulated in live HeLa and NIH3T3 cells. The two-photon-induced luminescence of the complex was retained after live cell internalization and can be observed by two-photon confocal microscopy. Its bioaccumulation properties enabled time-lapse imaging of the internalization process of the dye into living cells. Cytotoxicity of [Pt(L)Cl] to both tested cell lines was low, according to MTT assays, even at loadings as high as 20 times the dose concentration for imaging for 6 h.

  3. Multisite two-photon imaging of neurons on multielectrode arrays

    Science.gov (United States)

    Potter, Steve M.; Lukina, Natalia; Longmuir, Kenneth J.; Wu, Yan

    2001-04-01

    We wish to understand how neural systems store, recall, and process information. We are using cultured networks of cortical neurons grown on microelectrode arrays as a model system for studying the emergent properties of ensembles of living neurons. We have developed a 2-way communication interface between the cultured network and a computer- generated animal, the Neurally Controlled Animat. Neural activity is used to control the behavior of the Animat, and 2- photon time-lapse imaging is carried out in order to observe the morphological changes that might underlie changes in neural processing. The 2-photon microscope is ideal for repeated imaging over hours or days, with submicron resolution and little photodamage. We have designed a computer-controlled microscope stage that allows imaging several locations in sequence, in order to collect more image data. For the latest progress, see: http://www.caltech.edu/~pinelab/PotterGroup.htm.

  4. Near real-time imaging of molasses injections using time-lapse electrical geophysics at the Brandywine DRMO, Brandywine, Maryland

    Science.gov (United States)

    Versteeg, R. J.; Johnson, T.; Major, B.; Day-Lewis, F. D.; Lane, J. W.

    2010-12-01

    Enhanced bioremediation, which involves introduction of amendments to promote biodegradation, increasingly is used to accelerate cleanup of recalcitrant compounds and has been identified as the preferred remedial treatment at many contaminated sites. Although blind introduction of amendments can lead to sub-optimal or ineffective remediation, the distribution of amendment throughout the treatment zone is difficult to measure using conventional sampling. Because amendments and their degradation products commonly have electrical properties that differ from those of ambient soil, time-lapse electrical geophysical monitoring has the potential to verify amendment emplacement and distribution. In order for geophysical monitoring to be useful, however, results of the injection ideally should be accessible in near real time. In August 2010, we demonstrated the feasibility of near real-time, autonomous electrical geophysical monitoring of amendment injections at the former Defense Reutilization and Marketing Office (DRMO) in Brandywine, Maryland. Two injections of about 1000 gallons each of molasses, a widely used amendment for enhanced bioremediation, were monitored using measurements taken with borehole and surface electrodes. During the injections, multi-channel resistance data were recorded; data were transmitted to a server and processed using a parallel resistivity inversion code; and results in the form of time-lapse imagery subsequently were posted to a website. This process occurred automatically without human intervention. The resulting time-lapse imagery clearly showed the evolution of the molasses plume. The delay between measurements and online delivery of images was between 45 and 60 minutes, thus providing actionable information that could support decisions about field procedures and a check on whether amendment reached target zones. This experiment demonstrates the feasibility of using electrical imaging as a monitoring tool both during amendment emplacement

  5. Analysis of Zebrafish Kidney Development with Time-lapse Imaging Using a Dissecting Microscope Equipped for Optical Sectioning.

    Science.gov (United States)

    Perner, Birgit; Schnerwitzki, Danny; Graf, Michael; Englert, Christoph

    2016-04-07

    In order to understand organogenesis, the spatial and temporal alterations that occur during development of tissues need to be recorded. The method described here allows time-lapse analysis of normal and impaired kidney development in zebrafish embryos by using a fluorescence dissecting microscope equipped for structured illumination and z-stack acquisition. To visualize nephrogenesis, transgenic zebrafish (Tg(wt1b:GFP)) with fluorescently labeled kidney structures were used. Renal defects were triggered by injection of an antisense morpholino oligonucleotide against the Wilms tumor gene wt1a, a factor known to be crucial for kidney development. The advantage of the experimental setup is the combination of a zoom microscope with simple strategies for re-adjusting movements in x, y or z direction without additional equipment. To circumvent focal drift that is induced by temperature variations and mechanical vibrations, an autofocus strategy was applied instead of utilizing a usually required environmental chamber. In order to re-adjust the positional changes due to a xy-drift, imaging chambers with imprinted relocation grids were employed. In comparison to more complex setups for time-lapse recording with optical sectioning such as confocal laser scanning or light sheet microscopes, a zoom microscope is easy to handle. Besides, it offers dissecting microscope-specific benefits such as high depth of field and an extended working distance. The method to study organogenesis presented here can also be used with fluorescence stereo microscopes not capable of optical sectioning. Although limited for high-throughput, this technique offers an alternative to more complex equipment that is normally used for time-lapse recording of developing tissues and organ dynamics.

  6. Imaging rainfall infiltration processes with the time-lapse electrical resistivity imaging method

    Science.gov (United States)

    Jia, Zhengyuan; Jiang, Guoming; Zhang, Guibin; Zhang, Gang

    2017-04-01

    Electrical Resistivity Imaging (ERI) was carried out continuously for ten days to map the subsurface resistivity distribution along a potentially hazardous hillslope at the Jieshou Junior High School in Taoyuan, Taiwan. The inversions confirm the viability of ERI in tracking the movement of groundwater flow and rainfall infiltration by recording the variation of subsurface resistivity distribution. Meanwhile, relative-water-saturation (RWS) maps can be obtained from ERI images via Archie's Law, which provide a more intuitive reflection of the variation of subsurface rainfall infiltration and a more capable means of estimating the stability of a landslide body. What is more, we then found that the averaged RWS is significantly correlated with daily precipitation. Our observations indicate that real-time ERI is effective in monitoring subterraneous rainfall infiltration, and thereby in estimating the stability of a potential landslide body. When the agglomerate rainfall in the landslide slippage surface was infiltrated quickly without sustaining hydraulic pressure along the landslide slippage surface, the probability of landslides occurring was very low. On the contrary, the probability of landslides occurring could be increased due to the overpressure of pore fluids. Keywords Electrical Resistivity Imaging; Depth-of-Investigation; Archie's Law; Landslide Monitoring; Rainfall Infiltration; Preferential Path

  7. OPTICAL FLOW APPLIED TO TIME-LAPSE IMAGE SERIES TO ESTIMATE GLACIER MOTION IN THE SOUTHERN PATAGONIA ICE FIELD

    Directory of Open Access Journals (Sweden)

    E. Lannutti

    2016-06-01

    Full Text Available In this work, we assessed the feasibility of using optical flow to obtain the motion estimation of a glacier. In general, former investigations used to detect glacier changes involve solutions that require repeated observations which are many times based on extensive field work. Taking into account glaciers are usually located in geographically complex and hard to access areas, deploying time-lapse imaging sensors, optical flow may provide an efficient solution at good spatial and temporal resolution to describe mass motion. Several studies in computer vision and image processing community have used this method to detect large displacements. Therefore, we carried out a test of the proposed Large Displacement Optical Flow method at the Viedma Glacier, located at South Patagonia Icefield, Argentina. We collected monoscopic terrestrial time-lapse imagery, acquired by a calibrated camera at every 24 hour from April 2014 until April 2015. A filter based on temporal correlation and RGB color discretization between the images was applied to minimize errors related to changes in lighting, shadows, clouds and snow. This selection allowed discarding images that do not follow a sequence of similarity. Our results show a flow field in the direction of the glacier movement with acceleration in the terminus. We analyzed the errors between image pairs, and the matching generally appears to be adequate, although some areas show random gross errors related to the presence of changes in lighting. The proposed technique allowed the determination of glacier motion during one year, providing accurate and reliable motion data for subsequent analysis.

  8. A Modular and Affordable Time-Lapse Imaging and Incubation System Based on 3D-Printed Parts, a Smartphone, and Off-The-Shelf Electronics.

    Science.gov (United States)

    Hernández Vera, Rodrigo; Schwan, Emil; Fatsis-Kavalopoulos, Nikos; Kreuger, Johan

    2016-01-01

    Time-lapse imaging is a powerful tool for studying cellular dynamics and cell behavior over long periods of time to acquire detailed functional information. However, commercially available time-lapse imaging systems are expensive and this has limited a broader implementation of this technique in low-resource environments. Further, the availability of time-lapse imaging systems often present workflow bottlenecks in well-funded institutions. To address these limitations we have designed a modular and affordable time-lapse imaging and incubation system (ATLIS). The ATLIS enables the transformation of simple inverted microscopes into live cell imaging systems using custom-designed 3D-printed parts, a smartphone, and off-the-shelf electronic components. We demonstrate that the ATLIS provides stable environmental conditions to support normal cell behavior during live imaging experiments in both traditional and evaporation-sensitive microfluidic cell culture systems. Thus, the system presented here has the potential to increase the accessibility of time-lapse microscopy of living cells for the wider research community.

  9. A Modular and Affordable Time-Lapse Imaging and Incubation System Based on 3D-Printed Parts, a Smartphone, and Off-The-Shelf Electronics

    Science.gov (United States)

    Schwan, Emil; Fatsis-Kavalopoulos, Nikos; Kreuger, Johan

    2016-01-01

    Time-lapse imaging is a powerful tool for studying cellular dynamics and cell behavior over long periods of time to acquire detailed functional information. However, commercially available time-lapse imaging systems are expensive and this has limited a broader implementation of this technique in low-resource environments. Further, the availability of time-lapse imaging systems often present workflow bottlenecks in well-funded institutions. To address these limitations we have designed a modular and affordable time-lapse imaging and incubation system (ATLIS). The ATLIS enables the transformation of simple inverted microscopes into live cell imaging systems using custom-designed 3D-printed parts, a smartphone, and off-the-shelf electronic components. We demonstrate that the ATLIS provides stable environmental conditions to support normal cell behavior during live imaging experiments in both traditional and evaporation-sensitive microfluidic cell culture systems. Thus, the system presented here has the potential to increase the accessibility of time-lapse microscopy of living cells for the wider research community. PMID:28002463

  10. An effective assay for high cellular resolution time-lapse imaging of sensory placode formation and morphogenesis

    Directory of Open Access Journals (Sweden)

    Das Raman M

    2011-05-01

    Full Text Available Abstract Background The vertebrate peripheral nervous system contains sensory neurons that arise from ectodermal placodes. Placodal cells ingress to move inside the head to form sensory neurons of the cranial ganglia. To date, however, the process of placodal cell ingression and underlying cellular behavior are poorly understood as studies have relied upon static analyses on fixed tissues. Visualizing placodal cell behavior requires an ability to distinguish the surface ectoderm from the underlying mesenchyme. This necessitates high resolution imaging along the z-plane which is difficult to accomplish in whole embryos. To address this issue, we have developed an imaging system using cranial slices that allows direct visualization of placode formation. Results We demonstrate an effective imaging assay for capturing placode development at single cell resolution using chick embryonic tissue ex vivo. This provides the first time-lapse imaging of mitoses in the trigeminal placodal ectoderm, ingression, and intercellular contacts of placodal cells. Cell divisions with varied orientations were found in the placodal ectoderm all along the apical-basal axis. Placodal cells initially have short cytoplasmic processes during ingression as young neurons and mature over time to elaborate long axonal processes in the mesenchyme. Interestingly, the time-lapse imaging data reveal that these delaminating placodal neurons begin ingression early on from within the ectoderm, where they start to move and continue on to exit as individual or strings of neurons through common openings on the basal side of the epithelium. Furthermore, dynamic intercellular contacts are abundant among the delaminating placodal neurons, between these and the already delaminated cells, as well as among cells in the forming ganglion. Conclusions This new imaging assay provides a powerful method to analyze directly development of placode-derived sensory neurons and subsequent ganglia

  11. Imaging Live Drosophila Brain with Two-Photon Fluorescence Microscopy

    Science.gov (United States)

    Ahmed, Syeed Ehsan

    Two-photon fluorescence microscopy is an imaging technique which delivers distinct benefits for in vivo cellular and molecular imaging. Cyclic adenosine monophosphate (cAMP), a second messenger molecule, is responsible for triggering many physiological changes in neural system. However, the mechanism by which this molecule regulates responses in neuron cells is not yet clearly understood. When cAMP binds to a target protein, it changes the structure of that protein. Therefore, studying this molecular structure change with fluorescence resonance energy transfer (FRET) imaging can shed light on the cAMP functioning mechanism. FRET is a non-radiative dipole-dipole coupling which is sensitive to small distance change in nanometer scale. In this study we have investigated the effect of dopamine in cAMP dynamics in vivo. In our study two-photon fluorescence microscope was used for imaging mushroom bodies inside live Drosophila melanogaster brain and we developed a method for studying the change in cyclic AMP level.

  12. ViCAR: An Adaptive and Landmark-Free Registration of Time Lapse Image Data from Microfluidics Experiments

    Directory of Open Access Journals (Sweden)

    Georges Hattab

    2017-05-01

    Full Text Available In order to understand gene function in bacterial life cycles, time lapse bioimaging is applied in combination with different marker protocols in so called microfluidics chambers (i.e., a multi-well plate. In one experiment, a series of T images is recorded for one visual field, with a pixel resolution of 60 nm/px. Any (semi-automatic analysis of the data is hampered by a strong image noise, low contrast and, last but not least, considerable irregular shifts during the acquisition. Image registration corrects such shifts enabling next steps of the analysis (e.g., feature extraction or tracking. Image alignment faces two obstacles in this microscopic context: (a highly dynamic structural changes in the sample (i.e., colony growth and (b an individual data set-specific sample environment which makes the application of landmarks-based alignments almost impossible. We present a computational image registration solution, we refer to as ViCAR: (Visual (Cues based (Adaptive (Registration, for such microfluidics experiments, consisting of (1 the detection of particular polygons (outlined and segmented ones, referred to as visual cues, (2 the adaptive retrieval of three coordinates throughout different sets of frames, and finally (3 an image registration based on the relation of these points correcting both rotation and translation. We tested ViCAR with different data sets and have found that it provides an effective spatial alignment thereby paving the way to extract temporal features pertinent to each resulting bacterial colony. By using ViCAR, we achieved an image registration with 99.9% of image closeness, based on the average rmsd of 4.10−2 pixels, and superior results compared to a state of the art algorithm.

  13. Two-photon imaging and analysis of neural network dynamics

    International Nuclear Information System (INIS)

    Luetcke, Henry; Helmchen, Fritjof

    2011-01-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  14. Two-photon imaging and analysis of neural network dynamics

    Science.gov (United States)

    Lütcke, Henry; Helmchen, Fritjof

    2011-08-01

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  15. Two-photon imaging and analysis of neural network dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Luetcke, Henry; Helmchen, Fritjof [Brain Research Institute, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich (Switzerland)

    2011-08-15

    The glow of a starry night sky, the smell of a freshly brewed cup of coffee or the sound of ocean waves breaking on the beach are representations of the physical world that have been created by the dynamic interactions of thousands of neurons in our brains. How the brain mediates perceptions, creates thoughts, stores memories and initiates actions remains one of the most profound puzzles in biology, if not all of science. A key to a mechanistic understanding of how the nervous system works is the ability to measure and analyze the dynamics of neuronal networks in the living organism in the context of sensory stimulation and behavior. Dynamic brain properties have been fairly well characterized on the microscopic level of individual neurons and on the macroscopic level of whole brain areas largely with the help of various electrophysiological techniques. However, our understanding of the mesoscopic level comprising local populations of hundreds to thousands of neurons (so-called 'microcircuits') remains comparably poor. Predominantly, this has been due to the technical difficulties involved in recording from large networks of neurons with single-cell spatial resolution and near-millisecond temporal resolution in the brain of living animals. In recent years, two-photon microscopy has emerged as a technique which meets many of these requirements and thus has become the method of choice for the interrogation of local neural circuits. Here, we review the state-of-research in the field of two-photon imaging of neuronal populations, covering the topics of microscope technology, suitable fluorescent indicator dyes, staining techniques, and in particular analysis techniques for extracting relevant information from the fluorescence data. We expect that functional analysis of neural networks using two-photon imaging will help to decipher fundamental operational principles of neural microcircuits.

  16. Automated Functional Analysis of Astrocytes from Chronic Time-Lapse Calcium Imaging Data.

    Science.gov (United States)

    Wang, Yinxue; Shi, Guilai; Miller, David J; Wang, Yizhi; Wang, Congchao; Broussard, Gerard; Wang, Yue; Tian, Lin; Yu, Guoqiang

    2017-01-01

    Recent discoveries that astrocytes exert proactive regulatory effects on neural information processing and that they are deeply involved in normal brain development and disease pathology have stimulated broad interest in understanding astrocyte functional roles in brain circuit. Measuring astrocyte functional status is now technically feasible, due to recent advances in modern microscopy and ultrasensitive cell-type specific genetically encoded Ca 2+ indicators for chronic imaging. However, there is a big gap between the capability of generating large dataset via calcium imaging and the availability of sophisticated analytical tools for decoding the astrocyte function. Current practice is essentially manual, which not only limits analysis throughput but also risks introducing bias and missing important information latent in complex, dynamic big data. Here, we report a suite of computational tools, called Functional AStrocyte Phenotyping (FASP), for automatically quantifying the functional status of astrocytes. Considering the complex nature of Ca 2+ signaling in astrocytes and low signal to noise ratio, FASP is designed with data-driven and probabilistic principles, to flexibly account for various patterns and to perform robustly with noisy data. In particular, FASP explicitly models signal propagation, which rules out the applicability of tools designed for other types of data. We demonstrate the effectiveness of FASP using extensive synthetic and real data sets. The findings by FASP were verified by manual inspection. FASP also detected signals that were missed by purely manual analysis but could be confirmed by more careful manual examination under the guidance of automatic analysis. All algorithms and the analysis pipeline are packaged into a plugin for Fiji (ImageJ), with the source code freely available online at https://github.com/VTcbil/FASP.

  17. SEGMENTATION OF ENVIRONMENTAL TIME LAPSE IMAGE SEQUENCES FOR THE DETERMINATION OF SHORE LINES CAPTURED BY HAND-HELD SMARTPHONE CAMERAS

    Directory of Open Access Journals (Sweden)

    M. Kröhnert

    2017-09-01

    Full Text Available The relevance of globally environmental issues gains importance since the last years with still rising trends. Especially disastrous floods may cause in serious damage within very short times. Although conventional gauging stations provide reliable information about prevailing water levels, they are highly cost-intensive and thus just sparsely installed. Smartphones with inbuilt cameras, powerful processing units and low-cost positioning systems seem to be very suitable wide-spread measurement devices that could be used for geo-crowdsourcing purposes. Thus, we aim for the development of a versatile mobile water level measurement system to establish a densified hydrological network of water levels with high spatial and temporal resolution. This paper addresses a key issue of the entire system: the detection of running water shore lines in smartphone images. Flowing water never appears equally in close-range images even if the extrinsics remain unchanged. Its non-rigid behavior impedes the use of good practices for image segmentation as a prerequisite for water line detection. Consequently, we use a hand-held time lapse image sequence instead of a single image that provides the time component to determine a spatio-temporal texture image. Using a region growing concept, the texture is analyzed for immutable shore and dynamic water areas. Finally, the prevalent shore line is examined by the resultant shapes. For method validation, various study areas are observed from several distances covering urban and rural flowing waters with different characteristics. Future work provides a transformation of the water line into object space by image-to-geometry intersection.

  18. Segmentation of Environmental Time Lapse Image Sequences for the Determination of Shore Lines Captured by Hand-Held Smartphone Cameras

    Science.gov (United States)

    Kröhnert, M.; Meichsner, R.

    2017-09-01

    The relevance of globally environmental issues gains importance since the last years with still rising trends. Especially disastrous floods may cause in serious damage within very short times. Although conventional gauging stations provide reliable information about prevailing water levels, they are highly cost-intensive and thus just sparsely installed. Smartphones with inbuilt cameras, powerful processing units and low-cost positioning systems seem to be very suitable wide-spread measurement devices that could be used for geo-crowdsourcing purposes. Thus, we aim for the development of a versatile mobile water level measurement system to establish a densified hydrological network of water levels with high spatial and temporal resolution. This paper addresses a key issue of the entire system: the detection of running water shore lines in smartphone images. Flowing water never appears equally in close-range images even if the extrinsics remain unchanged. Its non-rigid behavior impedes the use of good practices for image segmentation as a prerequisite for water line detection. Consequently, we use a hand-held time lapse image sequence instead of a single image that provides the time component to determine a spatio-temporal texture image. Using a region growing concept, the texture is analyzed for immutable shore and dynamic water areas. Finally, the prevalent shore line is examined by the resultant shapes. For method validation, various study areas are observed from several distances covering urban and rural flowing waters with different characteristics. Future work provides a transformation of the water line into object space by image-to-geometry intersection.

  19. High contrast two-photon imaging of fingermarks

    Science.gov (United States)

    Stoltzfus, Caleb R.; Rebane, Aleksander

    2016-04-01

    Optically-acquired fingermarks are widely used as evidence across law enforcement agencies as well as in the courts of law. A common technique for visualizing latent fingermarks on nonporous surfaces consists of cyanoacrylate fuming of the fingerprint material, followed by impregnation with a fluorescent dye, which under ultra violet (UV) illumination makes the fingermarks visible and thus accessible for digital recording. However, there exist critical circumstances, when the image quality is compromised due to high background scattering, high auto-fluorescence of the substrate material, or other detrimental photo-physical and photo-chemical effects such as light-induced damage to the sample. Here we present a novel near-infrared (NIR), two-photon induced fluorescence imaging modality, which significantly enhances the quality of the fingermark images, especially when obtained from highly reflective and/or scattering surfaces, while at the same time reducing photo-damage to sensitive forensic samples.

  20. Whole brain imaging with Serial Two-Photon Tomography

    Directory of Open Access Journals (Sweden)

    Stephen P Amato

    2016-03-01

    Full Text Available Imaging entire mouse brains at submicron resolution has historically been a challenging undertaking and largely confined to the province of dedicated atlasing initiatives. The has limited systematic investigations into important areas of neuroscience, such as neural circuits, brain mapping and neurodegeneration. In this paper, we describe in detail Serial Two-Photon (STP tomography, a robust, reliable method for imaging entire brains with histological detail. We provide examples of how the basic methodology can be extended to other imaging modalities, such as optical coherence tomography, in order to provide unique contrast mechanisms. Furthermore we provide a survey of the research that STP tomography has enabled in the field of neuroscience, provide examples of how this technology enables quantitative whole brain studies, and discuss the current limitations of STP tomography-based approaches

  1. Fluorescent Pluronic nanodots for in vivo two-photon imaging

    International Nuclear Information System (INIS)

    Maurin, Mathieu; Vurth, Laeticia; Vial, Jean-Claude; Baldeck, Patrice; Stephan, Olivier; Marder, Seth R; Sanden, Boudewijn Van der

    2009-01-01

    We report the synthesis of new nanosized fluorescent probes based on bio-compatible polyethylene-polypropylene glycol (Pluronic) materials. In aqueous solution, mini-emulsification of Pluronic with a high fluorescent di-stryl benzene-modified derivative, exhibiting a two-photon absorption cross section as high as 2500 Goeppert-Mayer units at 800 nm, leads to nanoparticles exhibiting a hydrodynamic radius below 100 nm. We have demonstrated that these new probes with luminescence located in the spectral region of interest for bio-imaging (the yellow part of the visible spectrum) allow deep (500 μm) bio-imaging of the mice brain vasculature. The dose injected during our experiments is ten times lower when compared to the classical commercial rhodamine-B isothicyanate-Dextran system but gives similar results to homogeneous blood plasma staining. The mean fluorescent signal intensity stayed constant during more than 1 h.

  2. Two-Photon Ghost Image and Interference-Diffraction

    Science.gov (United States)

    Shih, Y. H.; Sergienko, A. V.; Pittman, T. B.; Strekalov, D. V.; Klyshko, D. N.

    1996-01-01

    One of the most surprising consequences of quantum mechanics is entanglement of two or more distance particles. The two-particle entangled state was mathematically formulated by Schrodinger. Based on this unusual quantum behavior, EPR defined their 'physical reality' and then asked the question: 'Can Quantum-Mechanical Description of Physical Reality Be Considered Complete?' One may not appreciate EPR's criterion of physical reality and insist that 'no elementary quantum phenomenon is a phenomenon until it is a recorded phenomenon'. Optical spontaneous parametric down conversion (SPDC) is the most effective mechanism to generate an EPR type entangled two-photon state. In SPDC, an optical beam, called the pump, is incident on a birefringent crystal. The pump is intense enough so that nonlinear effects lead to the conversion of pump photons into pairs of photons, historically called signal and idler. Technically, the SPDC is said to be type-1 or type-2, depending on whether the signal and idler beams have parallel or orthogonal polarization. The SPDC conversion efficiency is typically on the order of 10(exp -9) to 10(exp -11), depending on the SPDC nonlinear material. The signal and idler intensities are extremely low, only single photon detection devices can register them. The quantum entanglement nature of SPDC has been demonstrated in EPR-Bohm experiments and Bell's inequality measurements. The following two experiments were recently performed in our laboratory, which are more closely related to the original 1935 EPR gedankenezperiment. The first experiment is a two-photon optical imaging type experiment, which has been named 'ghost image' by the physics community. The signal and idler beams of SPDC are sent in different directions, so that the detection of the signal and idler photons can be performed by two distant photon counting detectors. An aperture object (mask) is placed in front of the signal photon detector and illuminated by the signal beam through a

  3. A Sparse Bayesian Imaging Technique for Efficient Recovery of Reservoir Channels With Time-Lapse Seismic Measurements

    KAUST Repository

    Sana, Furrukh

    2016-06-01

    Subsurface reservoir flow channels are characterized by high-permeability values and serve as preferred pathways for fluid propagation. Accurate estimation of their geophysical structures is thus of great importance for the oil industry. The ensemble Kalman filter (EnKF) is a widely used statistical technique for estimating subsurface reservoir model parameters. However, accurate reconstruction of the subsurface geological features with the EnKF is challenging because of the limited measurements available from the wells and the smoothing effects imposed by the \\\\ell _{2} -norm nature of its update step. A new EnKF scheme based on sparse domain representation was introduced by Sana et al. (2015) to incorporate useful prior structural information in the estimation process for efficient recovery of subsurface channels. In this paper, we extend this work in two ways: 1) investigate the effects of incorporating time-lapse seismic data on the channel reconstruction; and 2) explore a Bayesian sparse reconstruction algorithm with the potential ability to reduce the computational requirements. Numerical results suggest that the performance of the new sparse Bayesian based EnKF scheme is enhanced with the availability of seismic measurements, leading to further improvement in the recovery of flow channels structures. The sparse Bayesian approach further provides a computationally efficient framework for enforcing a sparse solution, especially with the possibility of using high sparsity rates through the inclusion of seismic data.

  4. A Sparse Bayesian Imaging Technique for Efficient Recovery of Reservoir Channels With Time-Lapse Seismic Measurements

    KAUST Repository

    Sana, Furrukh; Ravanelli, Fabio; Al-Naffouri, Tareq Y.; Hoteit, Ibrahim

    2016-01-01

    Subsurface reservoir flow channels are characterized by high-permeability values and serve as preferred pathways for fluid propagation. Accurate estimation of their geophysical structures is thus of great importance for the oil industry. The ensemble Kalman filter (EnKF) is a widely used statistical technique for estimating subsurface reservoir model parameters. However, accurate reconstruction of the subsurface geological features with the EnKF is challenging because of the limited measurements available from the wells and the smoothing effects imposed by the \\ell _{2} -norm nature of its update step. A new EnKF scheme based on sparse domain representation was introduced by Sana et al. (2015) to incorporate useful prior structural information in the estimation process for efficient recovery of subsurface channels. In this paper, we extend this work in two ways: 1) investigate the effects of incorporating time-lapse seismic data on the channel reconstruction; and 2) explore a Bayesian sparse reconstruction algorithm with the potential ability to reduce the computational requirements. Numerical results suggest that the performance of the new sparse Bayesian based EnKF scheme is enhanced with the availability of seismic measurements, leading to further improvement in the recovery of flow channels structures. The sparse Bayesian approach further provides a computationally efficient framework for enforcing a sparse solution, especially with the possibility of using high sparsity rates through the inclusion of seismic data.

  5. A-3 Construction Time Lapse

    Science.gov (United States)

    2009-01-01

    A time lapse from start to finish of steel erection for the 235-foot tall A-3 Test Stand. Ground work for the stand was broken in August 2008 and the final structural steel beam was placed April 9, 2009.

  6. Three-dimensional quantification of orthodontic root resorption with time-lapsed imaging of micro-computed tomography in a rodent model.

    Science.gov (United States)

    Yang, Chongshi; Zhang, Yuanyuan; Zhang, Yan; Fan, Yubo; Deng, Feng

    2015-01-01

    Despite various X-ray approaches have been widely used to monitor root resorption after orthodontic treatment, a non-invasive and accurate method is highly desirable for long-term follow up. The aim of this study was to build a non-invasive method to quantify longitudinal orthodontic root resorption with time-lapsed images of micro-computed tomography (micro-CT) in a rodent model. Twenty male Sprague Dawley (SD) rats (aged 6-8 weeks, weighing 180-220 g) were used in this study. A 25 g orthodontic force generated by nickel-titanium coil spring was applied to the right maxillary first molar for each rat, while contralateral first molar was severed as a control. Micro-CT scan was performed at day 0 (before orthodontic load) and days 3, 7, 14, and 28 after orthodontic load. Resorption of mesial root of maxillary first molars at bilateral sides was calculated from micro-CT images with registration algorithm via reconstruction, superimposition and partition operations. Obvious resorption of mesial root of maxillary first molar can be detected at day 14 and day 28 at orthodontic side. Most of the resorption occurred in the apical region at distal side and cervical region at mesiolingual side. Desirable development of molar root of rats was identified from day 0 to day 28 at control side. The development of root concentrated on apical region. This non-invasive 3D quantification method with registration algorithm can be used in longitudinal study of root resorption. Obvious root resorption in rat molar can be observed three-dimensionally at day 14 and day 28 after orthodontic load. This indicates that registration algorithm combined with time-lapsed images provides clinic potential application in detection and quantification of root contour.

  7. Automated profiling of individual cell-cell interactions from high-throughput time-lapse imaging microscopy in nanowell grids (TIMING).

    Science.gov (United States)

    Merouane, Amine; Rey-Villamizar, Nicolas; Lu, Yanbin; Liadi, Ivan; Romain, Gabrielle; Lu, Jennifer; Singh, Harjeet; Cooper, Laurence J N; Varadarajan, Navin; Roysam, Badrinath

    2015-10-01

    There is a need for effective automated methods for profiling dynamic cell-cell interactions with single-cell resolution from high-throughput time-lapse imaging data, especially, the interactions between immune effector cells and tumor cells in adoptive immunotherapy. Fluorescently labeled human T cells, natural killer cells (NK), and various target cells (NALM6, K562, EL4) were co-incubated on polydimethylsiloxane arrays of sub-nanoliter wells (nanowells), and imaged using multi-channel time-lapse microscopy. The proposed cell segmentation and tracking algorithms account for cell variability and exploit the nanowell confinement property to increase the yield of correctly analyzed nanowells from 45% (existing algorithms) to 98% for wells containing one effector and a single target, enabling automated quantification of cell locations, morphologies, movements, interactions, and deaths without the need for manual proofreading. Automated analysis of recordings from 12 different experiments demonstrated automated nanowell delineation accuracy >99%, automated cell segmentation accuracy >95%, and automated cell tracking accuracy of 90%, with default parameters, despite variations in illumination, staining, imaging noise, cell morphology, and cell clustering. An example analysis revealed that NK cells efficiently discriminate between live and dead targets by altering the duration of conjugation. The data also demonstrated that cytotoxic cells display higher motility than non-killers, both before and during contact. broysam@central.uh.edu or nvaradar@central.uh.edu Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  8. In vivo two-photon imaging of retina in rabbits and rats.

    Science.gov (United States)

    Jayabalan, Gopal Swamy; Wu, Yi-Kai; Bille, Josef F; Kim, Samuel; Mao, Xiao Wen; Gimbel, Howard V; Rauser, Michael E; Fan, Joseph T

    2018-01-01

    The purpose of this study was to evaluate the retina using near-infrared (NIR) two-photon scanning laser ophthalmoscopy. New Zealand white rabbits, albino rats, and brown Norway rats were used in this study. An autofluorescence image of the retina, including the retinal cells and its associated vasculatures was obtained by a real-time scan using the ophthalmoscope. Furthermore, the retinal vessels, nerve fiber layers and the non-pigmented retina were recorded with two-photon fluorescein angiography (FA); and the choroidal vasculatures were recorded using two-photon indocyanine green angiography (ICGA). Two-photon ICGA was achieved by exciting a second singlet state at ∼398 nm. Simultaneous two-photon FA and two-photon ICGA were performed to characterize the retinal and choroidal vessels with a single injection. The minimum laser power threshold required to elicit two-photon fluorescence was determined. The two-photon ophthalmoscope could serve as a promising tool to detect and monitor the disease progression in animal models. Moreover, these high-resolution images of retinal and choroidal vessels can be acquired in a real-time scan with a single light source, requiring no additional filters for FA or ICGA. The combination of FA and ICGA using the two-photon ophthalmoscope will help researchers to characterize the retinal diseases in animal models, and also to classify the types (classic, occult or mixed) of choroidal neovascularization (CNV) in macular degeneration. Furthermore, the prototype can be adapted to image the retina of rodents and rabbits. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. New algorithm to determine true colocalization in combination with image restoration and time-lapse confocal microscopy to MAP kinases in mitochondria.

    Directory of Open Access Journals (Sweden)

    Jorge Ignacio Villalta

    Full Text Available The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized markers and posterior evaluation of the signal colocalization. Rigorous study of colocalization requires statistical analysis of the data, albeit yet no single technique has been established as a standard method. Indeed, the few methods currently available are only accurate in images with particular characteristics. Here, we introduce a new algorithm to automatically obtain the true colocalization between images that is suitable for a wide variety of biological situations. To proceed, the algorithm contemplates the individual contribution of each pixel's fluorescence intensity in a pair of images to the overall Pearsońs correlation and Manders' overlap coefficients. The accuracy and reliability of the algorithm was validated on both simulated and real images that reflected the characteristics of a range of biological samples. We used this algorithm in combination with image restoration by deconvolution and time-lapse confocal microscopy to address the localization of MEK1 in the mitochondria of different cell lines. Appraising the previously described behavior of Akt1 corroborated the reliability of the combined use of these techniques. Together, the present work provides a novel statistical approach to accurately and reliably determine the colocalization in a variety of biological images.

  10. Images of photoreceptors in living primate eyes using adaptive optics two-photon ophthalmoscopy

    Science.gov (United States)

    Hunter, Jennifer J.; Masella, Benjamin; Dubra, Alfredo; Sharma, Robin; Yin, Lu; Merigan, William H.; Palczewska, Grazyna; Palczewski, Krzysztof; Williams, David R.

    2011-01-01

    In vivo two-photon imaging through the pupil of the primate eye has the potential to become a useful tool for functional imaging of the retina. Two-photon excited fluorescence images of the macaque cone mosaic were obtained using a fluorescence adaptive optics scanning laser ophthalmoscope, overcoming the challenges of a low numerical aperture, imperfect optics of the eye, high required light levels, and eye motion. Although the specific fluorophores are as yet unknown, strong in vivo intrinsic fluorescence allowed images of the cone mosaic. Imaging intact ex vivo retina revealed that the strongest two-photon excited fluorescence signal comes from the cone inner segments. The fluorescence response increased following light stimulation, which could provide a functional measure of the effects of light on photoreceptors. PMID:21326644

  11. Two-Photon Autofluorescence Imaging Reveals Cellular Structures Throughout the Retina of the Living Primate Eye.

    Science.gov (United States)

    Sharma, Robin; Williams, David R; Palczewska, Grazyna; Palczewski, Krzysztof; Hunter, Jennifer J

    2016-02-01

    Although extrinsic fluorophores can be introduced to label specific cell types in the retina, endogenous fluorophores, such as NAD(P)H, FAD, collagen, and others, are present in all retinal layers. These molecules are a potential source of optical contrast and can enable noninvasive visualization of all cellular layers. We used a two-photon fluorescence adaptive optics scanning light ophthalmoscope (TPF-AOSLO) to explore the native autofluorescence of various cell classes spanning several layers in the unlabeled retina of a living primate eye. Three macaques were imaged on separate occasions using a custom TPF-AOSLO. Two-photon fluorescence was evoked by pulsed light at 730 and 920 nm excitation wavelengths, while fluorescence emission was collected in the visible range from several retinal layers and different locations. Backscattered light was recorded simultaneously in confocal modality and images were postprocessed to remove eye motion. All retinal layers yielded two-photon signals and the heterogeneous distribution of fluorophores provided optical contrast. Several structural features were observed, such as autofluorescence from vessel walls, Müller cell processes in the nerve fibers, mosaics of cells in the ganglion cell and other nuclear layers of the inner retina, as well as photoreceptor and RPE layers in the outer retina. This in vivo survey of two-photon autofluorescence throughout the primate retina demonstrates a wider variety of structural detail in the living eye than is available through conventional imaging methods, and broadens the use of two-photon imaging of normal and diseased eyes.

  12. Two-photon calcium imaging in mice navigating a virtual reality environment.

    Science.gov (United States)

    Leinweber, Marcus; Zmarz, Pawel; Buchmann, Peter; Argast, Paul; Hübener, Mark; Bonhoeffer, Tobias; Keller, Georg B

    2014-02-20

    In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically identified cells during behavior(1-6). Here we describe methods to perform two-photon imaging in mouse cortex while the animal navigates a virtual reality environment. We focus on the aspects of the experimental procedures that are key to imaging in a behaving animal in a brightly lit virtual environment. The key problems that arise in this experimental setup that we here address are: minimizing brain motion related artifacts, minimizing light leak from the virtual reality projection system, and minimizing laser induced tissue damage. We also provide sample software to control the virtual reality environment and to do pupil tracking. With these procedures and resources it should be possible to convert a conventional two-photon microscope for use in behaving mice.

  13. Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging

    NARCIS (Netherlands)

    Uzunbajakava, N.; Otto, Cornelis

    2003-01-01

    We demonstrate a confocal optical microscope that combines cw two-photon-excited fluorescence microscopy with confocal Raman microscopy. With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous

  14. Applications of two-photon fluorescence microscopy in deep-tissue imaging

    Science.gov (United States)

    Dong, Chen-Yuan; Yu, Betty; Hsu, Lily L.; Kaplan, Peter D.; Blankschstein, D.; Langer, Robert; So, Peter T. C.

    2000-07-01

    Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.

  15. Imaging subsurface migration of dissolved CO2 in a shallow aquifer using 3-D time-lapse electrical resistivity tomography

    DEFF Research Database (Denmark)

    Auken, Esben; Doetsch, Joseph; Fiandaca, Gianluca

    2014-01-01

    Contamination of groundwater by leaking CO2 is a potential risk of carbon sequestration. With the help of a field experiment in western Denmark, we investigate to what extent surface electrical resistivity tomography (ERT) can detect and image dissolved CO2 in a shallow aquifer. For this purpose...... the injection start. During these 120days, the CO2 migrates about 25m in the expected groundwater flow direction. Water electrical conductivity (EC) sampling using small screens in 29 wells allows for very good verification of the ERT results. Water EC and ERT results generally agree very well, with the water...

  16. Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography

    Science.gov (United States)

    Patalay, Rakesh; Talbot, Clifford; Alexandrov, Yuriy; Munro, Ian; Breunig, Hans Georg; König, Karsten; Warren, Sean; Neil, Mark A. A.; French, Paul M. W.; Chu, Anthony; Stamp, Gordon W.; Dunsby, Christopher

    2011-07-01

    Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential.

  17. Time-lapse imaging of fault properties at seismogenic depth using repeating earthquakes, active sources and seismic ambient noise

    Science.gov (United States)

    Cheng, Xin

    2009-12-01

    The time-varying stress field of fault systems at seismogenic depths plays the mort important role in controlling the sequencing and nucleation of seismic events. Using seismic observations from repeating earthquakes, controlled active sources and seismic ambient noise, five studies at four different fault systems across North America, Central Japan, North and mid-West China are presented to describe our efforts to measure such time dependent structural properties. Repeating and similar earthquakes are hunted and analyzed to study the post-seismic fault relaxation at the aftershock zone of the 1984 M 6.8 western Nagano and the 1976 M 7.8 Tangshan earthquakes. The lack of observed repeating earthquakes at western Nagano is attributed to the absence of a well developed weak fault zone, suggesting that the fault damage zone has been almost completely healed. In contrast, the high percentage of similar and repeating events found at Tangshan suggest the existence of mature fault zones characterized by stable creep under steady tectonic loading. At the Parkfield region of the San Andreas Fault, repeating earthquake clusters and chemical explosions are used to construct a scatterer migration image based on the observation of systematic temporal variations in the seismic waveforms across the occurrence time of the 2004 M 6 Parkfield earthquake. Coseismic fluid charge or discharge in fractures caused by the Parkfield earthquake is used to explain the observed seismic scattering properties change at depth. In the same region, a controlled source cross-well experiment conducted at SAFOD pilot and main holes documents two large excursions in the travel time required for a shear wave to travel through the rock along a fixed pathway shortly before two rupture events, suggesting that they may be related to pre-rupture stress induced changes in crack properties. At central China, a tomographic inversion based on the theory of seismic ambient noise and coda wave interferometry

  18. Revelation of Different Nanoparticle-Uptake Behavior in Two Standard Cell Lines NIH/3T3 and A549 by Flow Cytometry and Time-Lapse Imaging

    Directory of Open Access Journals (Sweden)

    André Jochums

    2017-07-01

    Full Text Available The uptake of nanomaterials into different cell types is a central pharmacological issue for the determination of nanotoxicity as well as for the development of drug delivery strategies. Most responses of the cells depend on their intracellular interactions with nanoparticles (NPs. Uptake behavior can be precisely investigated in vitro, with sensitive high throughput methods such as flow cytometry. In this study, we investigated two different standard cell lines, human lung carcinoma (A549 and mouse fibroblast (NIH/3T3 cells, regarding their uptake behavior of titanium dioxide NPs. Cells were incubated with different concentrations of TiO2 NPs and samples were taken at certain time points to compare the uptake kinetics of both cell lines. Samples were analyzed with the help of flow cytometry by studying changes in the side and forward scattering signal. To additionally enable a detection via fluorescence, NPs were labeled with the fluorescent dye fluorescein isothiocyanate (FITC and propidium iodide (PI. We found that NIH/3T3 cells take up the studied NPs more efficiently than A549 cells. These findings were supported by time-lapse microscopic imaging of the cells incubated with TiO2 NPs. Our results confirm that the uptake behavior of individual cell types has to be considered before interpreting any results of nanomaterial studies.

  19. Near-IR Two-Photon Fluorescent Sensor for K(+) Imaging in Live Cells.

    Science.gov (United States)

    Sui, Binglin; Yue, Xiling; Kim, Bosung; Belfield, Kevin D

    2015-08-19

    A new two-photon excited fluorescent K(+) sensor is reported. The sensor comprises three moieties, a highly selective K(+) chelator as the K(+) recognition unit, a boron-dipyrromethene (BODIPY) derivative modified with phenylethynyl groups as the fluorophore, and two polyethylene glycol chains to afford water solubility. The sensor displays very high selectivity (>52-fold) in detecting K(+) over other physiological metal cations. Upon binding K(+), the sensor switches from nonfluorescent to highly fluorescent, emitting red to near-IR (NIR) fluorescence. The sensor exhibited a good two-photon absorption cross section, 500 GM at 940 nm. Moreover, it is not sensitive to pH in the physiological pH range. Time-dependent cell imaging studies via both one- and two-photon fluorescence microscopy demonstrate that the sensor is suitable for dynamic K(+) sensing in living cells.

  20. Scanless functional imaging of hippocampal networks using patterned two-photon illumination through GRIN lenses

    KAUST Repository

    Moretti, Claudio

    2016-09-12

    Patterned illumination through the phase modulation of light is increasingly recognized as a powerful tool to investigate biological tissues in combination with two-photon excitation and light-sensitive molecules. However, to date two-photon patterned illumination has only been coupled to traditional microscope objectives, thus limiting the applicability of these methods to superficial biological structures. Here, we show that phase modulation can be used to efficiently project complex two-photon light patterns, including arrays of points and large shapes, in the focal plane of graded index (GRIN) lenses. Moreover, using this approach in combination with the genetically encoded calcium indicator GCaMP6, we validate our system performing scanless functional imaging in rodent hippocampal networks in vivo ~1.2 mm below the brain surface. Our results open the way to the application of patterned illumination approaches to deep regions of highly scattering biological tissues, such as the mammalian brain.

  1. Gold Core Mesoporous Organosilica Shell Degradable Nanoparticles for Two-Photon Imaging and Gemcitabine Monophosphate Delivery

    KAUST Repository

    Rhamani, Saher; Chaix, Arnaud; Aggad, Dina; Hoang, Phuong Mai; Moosa, Basem; Garcia, Marcel; Gary-Bobo, Magali; Charnay, Clarence; Almalik, Abdulaziz; Durand, Jean-Olivier; Khashab, Niveen M.

    2017-01-01

    The synthesis of gold core degradable mesoporous organosilica shell nanoparticles is described. The nanopaticles were very efficient for two-photon luminescence imaging of cancer cells and for in vitro gemcitabine monophosphate delivery, allowing promising theranostic applications in the nanomedicine field.

  2. Gold Core Mesoporous Organosilica Shell Degradable Nanoparticles for Two-Photon Imaging and Gemcitabine Monophosphate Delivery

    KAUST Repository

    Rhamani, Saher

    2017-09-12

    The synthesis of gold core degradable mesoporous organosilica shell nanoparticles is described. The nanopaticles were very efficient for two-photon luminescence imaging of cancer cells and for in vitro gemcitabine monophosphate delivery, allowing promising theranostic applications in the nanomedicine field.

  3. Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)

    Science.gov (United States)

    Song, Alexander; Charles, Adam S.; Koay, Sue Ann; Gauthier, Jeff L.; Thiberge, Stephan Y.; Pillow, Jonathan W.; Tank, David W.

    2017-01-01

    Two-photon laser scanning microscopy of calcium dynamics using fluorescent indicators is a widely used imaging method for large scale recording of neural activity in vivo. Here we introduce volumetric Two-photon Imaging of Neurons using Stereoscopy (vTwINS), a volumetric calcium imaging method that employs an elongated, V-shaped point spread function to image a 3D brain volume. Single neurons project to spatially displaced “image pairs” in the resulting 2D image, and the separation distance between images is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a novel orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrate vTwINS by imaging neural population activity in mouse primary visual cortex and hippocampus. Our results demonstrate that vTwINS provides an effective method for volumetric two-photon calcium imaging that increases the number of neurons recorded while maintaining a high frame-rate. PMID:28319111

  4. 4-D imaging of seepage in earthen embankments with time-lapse inversion of self-potential data constrained by acoustic emissions localization

    Science.gov (United States)

    Rittgers, J. B.; Revil, A.; Planes, T.; Mooney, M. A.; Koelewijn, A. R.

    2015-02-01

    New methods are required to combine the information contained in the passive electrical and seismic signals to detect, localize and monitor hydromechanical disturbances in porous media. We propose a field experiment showing how passive seismic and electrical data can be combined together to detect a preferential flow path associated with internal erosion in a Earth dam. Continuous passive seismic and electrical (self-potential) monitoring data were recorded during a 7-d full-scale levee (earthen embankment) failure test, conducted in Booneschans, Netherlands in 2012. Spatially coherent acoustic emissions events and the development of a self-potential anomaly, associated with induced concentrated seepage and internal erosion phenomena, were identified and imaged near the downstream toe of the embankment, in an area that subsequently developed a series of concentrated water flows and sand boils, and where liquefaction of the embankment toe eventually developed. We present a new 4-D grid-search algorithm for acoustic emissions localization in both time and space, and the application of the localization results to add spatially varying constraints to time-lapse 3-D modelling of self-potential data in the terms of source current localization. Seismic signal localization results are utilized to build a set of time-invariant yet spatially varying model weights used for the inversion of the self-potential data. Results from the combination of these two passive techniques show results that are more consistent in terms of focused ground water flow with respect to visual observation on the embankment. This approach to geophysical monitoring of earthen embankments provides an improved approach for early detection and imaging of the development of embankment defects associated with concentrated seepage and internal erosion phenomena. The same approach can be used to detect various types of hydromechanical disturbances at larger scales.

  5. Multiscale vision model for event detection and reconstruction in two-photon imaging data

    DEFF Research Database (Denmark)

    Brazhe, Alexey; Mathiesen, Claus; Lind, Barbara Lykke

    2014-01-01

    on a modified multiscale vision model, an object detection framework based on the thresholding of wavelet coefficients and hierarchical trees of significant coefficients followed by nonlinear iterative partial object reconstruction, for the analysis of two-photon calcium imaging data. The framework is discussed...... of the multiscale vision model is similar in the denoising, but provides a better segmenation of the image into meaningful objects, whereas other methods need to be combined with dedicated thresholding and segmentation utilities....

  6. Two-photon excited autofluorescence imaging of human retinal pigment epithelial cells

    Science.gov (United States)

    Han, Meng; Blindewald-Wittich, Almut; Holz, Frank G.; Giese, Günter; Niemz, Markolf H.; Snyder, Sarah; Sun, Hui; Yu, Jiayi; Agopov, Michael; La Schiazza, Olivier; Bille, Josef F.

    2006-01-01

    Degeneration of retinal pigment epithelial (RPE) cells severely impairs the visual function of retina photoreceptors. However, little is known about the events that trigger the death of RPE cells at the subcellular level. Two-photon excited autofluorescence (TPEF) imaging of RPE cells proves to be well suited to investigate both the morphological and the spectral characteristics of the human RPE cells. The dominant fluorophores of autofluorescence derive from lipofuscin (LF) granules that accumulate in the cytoplasm of the RPE cells with increasing age. Spectral TPEF imaging reveals the existence of abnormal LF granules with blue shifted autofluorescence in RPE cells of aging patients and brings new insights into the complicated composition of the LF granules. Based on a proposed two-photon laser scanning ophthalmoscope, TPEF imaging of the living retina may be valuable for diagnostic and pathological studies of age related eye diseases.

  7. Diagnosis of basal cell carcinoma by two photon excited fluorescence combined with lifetime imaging

    Science.gov (United States)

    Fan, Shunping; Peng, Xiao; Liu, Lixin; Liu, Shaoxiong; Lu, Yuan; Qu, Junle

    2014-02-01

    Basal cell carcinoma (BCC) is the most common type of human skin cancer. The traditional diagnostic procedure of BCC is histological examination with haematoxylin and eosin staining of the tissue biopsy. In order to reduce complexity of the diagnosis procedure, a number of noninvasive optical methods have been applied in skin examination, for example, multiphoton tomography (MPT) and fluorescence lifetime imaging microscopy (FLIM). In this study, we explored two-photon optical tomography of human skin specimens using two-photon excited autofluorescence imaging and FLIM. There are a number of naturally endogenous fluorophores in skin sample, such as keratin, melanin, collagen, elastin, flavin and porphyrin. Confocal microscopy was used to obtain structures of the sample. Properties of epidermic and cancer cells were characterized by fluorescence emission spectra, as well as fluorescence lifetime imaging. Our results show that two-photon autofluorescence lifetime imaging can provide accurate optical biopsies with subcellular resolution and is potentially a quantitative optical diagnostic method in skin cancer diagnosis.

  8. Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators.

    Science.gov (United States)

    Chamberland, Simon; Yang, Helen H; Pan, Michael M; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying; Salesse, Charleen; Wu, Haodi; Wu, Joseph C; Clandinin, Thomas R; Toth, Katalin; Lin, Michael Z; St-Pierre, François

    2017-07-27

    Monitoring voltage dynamics in defined neurons deep in the brain is critical for unraveling the function of neuronal circuits but is challenging due to the limited performance of existing tools. In particular, while genetically encoded voltage indicators have shown promise for optical detection of voltage transients, many indicators exhibit low sensitivity when imaged under two-photon illumination. Previous studies thus fell short of visualizing voltage dynamics in individual neurons in single trials. Here, we report ASAP2s, a novel voltage indicator with improved sensitivity. By imaging ASAP2s using random-access multi-photon microscopy, we demonstrate robust single-trial detection of action potentials in organotypic slice cultures. We also show that ASAP2s enables two-photon imaging of graded potentials in organotypic slice cultures and in Drosophila . These results demonstrate that the combination of ASAP2s and fast two-photon imaging methods enables detection of neural electrical activity with subcellular spatial resolution and millisecond-timescale precision.

  9. Two-photon NADH imaging exposes boundaries of oxygen diffusion in cortical vascular supply regions

    OpenAIRE

    Kasischke, Karl A; Lambert, Elton M; Panepento, Ben; Sun, Anita; Gelbard, Harris A; Burgess, Robert W; Foster, Thomas H; Nedergaard, Maiken

    2010-01-01

    Oxygen transport imposes a possible constraint on the brain's ability to sustain variable metabolic demands, but oxygen diffusion in the cerebral cortex has not yet been observed directly. We show that concurrent two-photon fluorescence imaging of endogenous nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation exposes well-defined boundaries of tissue oxygen diffusion in the mouse cortex. The NADH fluorescence increases rapidly over a narrow, very low pO2 range with a p ...

  10. Additive controlled synthesis of gold nanorods (GNRs) for two-photon luminescence imaging of cancer cells

    International Nuclear Information System (INIS)

    Zhu Jing; Roy, Indrajit; Hu Rui; Ding Hong; Zhao Lingling; He, Guang S; Prasad, Paras N; Yong, Ken-Tye; Swihart, Mark T; Cui Yiping

    2010-01-01

    Gold nanorods (GNRs) with a longitudinal surface plasmon resonance peak that is tunable from 600 to 1100 nm have been fabricated in a cetyl trimethylammoniumbromide (CTAB) micellar medium using hydrochloric acid and silver nitrate as additives to control their shape and size. By manipulating the concentrations of silver nitrate and hydrochloric acid, the aspect ratio of the GNRs was reliably and reproducibly tuned from 2.5 to 8. The GNRs were first coated with polyelectrolyte multilayers and then bioconjugated to transferrin (Tf) to target pancreatic cancer cells. Two-photon imaging excited from the bioconjugated GNRs demonstrated receptor-mediated uptake of the bioconjugates into Panc-1 cells, overexpressing the transferrin receptor (TfR). The bioconjugated GNR formulation exhibited very low toxicity, suggesting that it is biocompatible and potentially suitable for targeted two-photon bioimaging.

  11. Additive controlled synthesis of gold nanorods (GNRs) for two-photon luminescence imaging of cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Jing; Roy, Indrajit; Hu Rui; Ding Hong; Zhao Lingling; He, Guang S; Prasad, Paras N [Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY 14260-4200 (United States); Yong, Ken-Tye [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Swihart, Mark T [Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Buffalo, NY 14260-4200 (United States); Cui Yiping, E-mail: ktyong@ntu.edu.sg, E-mail: cyp@seu.edu.cn, E-mail: pnprasad@buffalo.edu [Advanced Photonics Center, School of Electronic Science and Engineering, Southeast University, Nanjing 210096 (China)

    2010-07-16

    Gold nanorods (GNRs) with a longitudinal surface plasmon resonance peak that is tunable from 600 to 1100 nm have been fabricated in a cetyl trimethylammoniumbromide (CTAB) micellar medium using hydrochloric acid and silver nitrate as additives to control their shape and size. By manipulating the concentrations of silver nitrate and hydrochloric acid, the aspect ratio of the GNRs was reliably and reproducibly tuned from 2.5 to 8. The GNRs were first coated with polyelectrolyte multilayers and then bioconjugated to transferrin (Tf) to target pancreatic cancer cells. Two-photon imaging excited from the bioconjugated GNRs demonstrated receptor-mediated uptake of the bioconjugates into Panc-1 cells, overexpressing the transferrin receptor (TfR). The bioconjugated GNR formulation exhibited very low toxicity, suggesting that it is biocompatible and potentially suitable for targeted two-photon bioimaging.

  12. Additive controlled synthesis of gold nanorods (GNRs) for two-photon luminescence imaging of cancer cells

    Science.gov (United States)

    Zhu, Jing; Yong, Ken-Tye; Roy, Indrajit; Hu, Rui; Ding, Hong; Zhao, Lingling; Swihart, Mark T.; He, Guang S.; Cui, Yiping; Prasad, Paras N.

    2010-07-01

    Gold nanorods (GNRs) with a longitudinal surface plasmon resonance peak that is tunable from 600 to 1100 nm have been fabricated in a cetyl trimethylammoniumbromide (CTAB) micellar medium using hydrochloric acid and silver nitrate as additives to control their shape and size. By manipulating the concentrations of silver nitrate and hydrochloric acid, the aspect ratio of the GNRs was reliably and reproducibly tuned from 2.5 to 8. The GNRs were first coated with polyelectrolyte multilayers and then bioconjugated to transferrin (Tf) to target pancreatic cancer cells. Two-photon imaging excited from the bioconjugated GNRs demonstrated receptor-mediated uptake of the bioconjugates into Panc-1 cells, overexpressing the transferrin receptor (TfR). The bioconjugated GNR formulation exhibited very low toxicity, suggesting that it is biocompatible and potentially suitable for targeted two-photon bioimaging.

  13. Efficient multi-site two-photon functional imaging of neuronal circuits.

    Science.gov (United States)

    Castanares, Michael Lawrence; Gautam, Vini; Drury, Jack; Bachor, Hans; Daria, Vincent R

    2016-12-01

    Two-photon imaging using high-speed multi-channel detectors is a promising approach for optical recording of cellular membrane dynamics at multiple sites. A main bottleneck of this technique is the limited number of photons captured within a short exposure time (~1ms). Here, we implement temporal gating to improve the two-photon fluorescence yield from holographically projected multiple foci whilst maintaining a biologically safe incident average power. We observed up to 6x improvement in the signal-to-noise ratio (SNR) in Fluorescein and cultured hippocampal neurons showing evoked calcium transients. With improved SNR, we could pave the way to achieving multi-site optical recording of fluorogenic probes with response times in the order of ~1ms.

  14. Deep brain two-photon NIR fluorescence imaging for study of Alzheimer's disease

    Science.gov (United States)

    Chen, Congping; Liang, Zhuoyi; Zhou, Biao; Ip, Nancy Y.; Qu, Jianan Y.

    2018-02-01

    Amyloid depositions in the brain represent the characteristic hallmarks of Alzheimer's disease (AD) pathology. The abnormal accumulation of extracellular amyloid-beta (Aβ) and resulting toxic amyloid plaques are considered to be responsible for the clinical deficits including cognitive decline and memory loss. In vivo two-photon fluorescence imaging of amyloid plaques in live AD mouse model through a chronic imaging window (thinned skull or craniotomy) provides a mean to greatly facilitate the study of the pathological mechanism of AD owing to its high spatial resolution and long-term continuous monitoring. However, the imaging depth for amyloid plaques is largely limited to upper cortical layers due to the short-wavelength fluorescence emission of commonly used amyloid probes. In this work, we reported that CRANAD-3, a near-infrared (NIR) probe for amyloid species with excitation wavelength at 900 nm and emission wavelength around 650 nm, has great advantages over conventionally used probes and is well suited for twophoton deep imaging of amyloid plaques in AD mouse brain. Compared with a commonly used MeO-X04 probe, the imaging depth of CRANAD-3 is largely extended for open skull cranial window. Furthermore, by using two-photon excited fluorescence spectroscopic imaging, we characterized the intrinsic fluorescence of the "aging pigment" lipofuscin in vivo, which has distinct spectra from CRANAD-3 labeled plaques. This study reveals the unique potential of NIR probes for in vivo, high-resolution and deep imaging of brain amyloid in Alzheimer's disease.

  15. Application of quantitative time-lapse imaging (QTLI) for evaluation of Mrp2-based drug–drug interaction induced by liver metabolites

    Energy Technology Data Exchange (ETDEWEB)

    Nakanishi, Takeo; Ikenaga, Miho; Fukuda, Hajime; Matsunaga, Norikazu; Tamai, Ikumi, E-mail: tamai@p.kanazawa-w.ac.jp

    2012-09-01

    We previously reported a quantitative time-lapse imaging (QTLI)-based analysis method to assess drug–drug interactions (DDI) at multidrug resistance-associated protein 2 (Mrp2) in rat sandwich-cultured hepatocyte (SCH) system, utilizing the fluorescent Mrp2 substrate, 5-(and 6)-carboxy-2′,7′-dichlorofluorescein (CDF). Here, we aimed to examine the feasibility of using QTLI to evaluate DDI involving drug metabolite(s) generated in hepatocytes. We used estradiol (E2) and bilirubin as model compounds; both are not substrates of MRP2, whereas their hepatic metabolites, estradiol-17β-glucuronide (E17G) or bilirubin glucuronides, are known to be its substrates as well as inhibitors. When rat SCHs were pre-exposed with E2, fluorescence of CDF accumulated in bile canaliculi decreased depending upon both the duration of pre-exposure and the concentration of extracellular E2. The decrease corresponded with the increase in intracellular concentration of E17G in hepatocytes. Furthermore, cytotoxicity of vinblastine, a substrate of MRP2, was enhanced in SCHs treated with E2. Similarly, CDF accumulated in bile canaliculi was significantly reduced in rat SCHs pre-exposed with bilirubin. In conclusion, these results suggest that phase II biotransformation of a competitor is reflected in alteration of MRP2-mediated CDF transport detected in QTLI. The QTLI might provide a convenient platform to evaluate transporter-based DDIs involving hepatic metabolites of drug candidates without the need to identify the metabolites. -- Highlights: ► Mrp2-mediated CDF transport is inhibited by E2, but not E17G in vesicle study. ► Both E2 and E17G do not compromise CDF formation from CDFDA in hepatocytes. ► CDF accumulation in bile canaliculi is inhibited by E2 or E17G in QTLI. ► Increasing exposure to E2 decreases CDF accumulation in bile canaliculi in QTLI. ► QTLI is feasible to assess Mrp2-based DDI involving drug metabolite in hepatocytes.

  16. Application of quantitative time-lapse imaging (QTLI) for evaluation of Mrp2-based drug–drug interaction induced by liver metabolites

    International Nuclear Information System (INIS)

    Nakanishi, Takeo; Ikenaga, Miho; Fukuda, Hajime; Matsunaga, Norikazu; Tamai, Ikumi

    2012-01-01

    We previously reported a quantitative time-lapse imaging (QTLI)-based analysis method to assess drug–drug interactions (DDI) at multidrug resistance-associated protein 2 (Mrp2) in rat sandwich-cultured hepatocyte (SCH) system, utilizing the fluorescent Mrp2 substrate, 5-(and 6)-carboxy-2′,7′-dichlorofluorescein (CDF). Here, we aimed to examine the feasibility of using QTLI to evaluate DDI involving drug metabolite(s) generated in hepatocytes. We used estradiol (E2) and bilirubin as model compounds; both are not substrates of MRP2, whereas their hepatic metabolites, estradiol-17β-glucuronide (E17G) or bilirubin glucuronides, are known to be its substrates as well as inhibitors. When rat SCHs were pre-exposed with E2, fluorescence of CDF accumulated in bile canaliculi decreased depending upon both the duration of pre-exposure and the concentration of extracellular E2. The decrease corresponded with the increase in intracellular concentration of E17G in hepatocytes. Furthermore, cytotoxicity of vinblastine, a substrate of MRP2, was enhanced in SCHs treated with E2. Similarly, CDF accumulated in bile canaliculi was significantly reduced in rat SCHs pre-exposed with bilirubin. In conclusion, these results suggest that phase II biotransformation of a competitor is reflected in alteration of MRP2-mediated CDF transport detected in QTLI. The QTLI might provide a convenient platform to evaluate transporter-based DDIs involving hepatic metabolites of drug candidates without the need to identify the metabolites. -- Highlights: ► Mrp2-mediated CDF transport is inhibited by E2, but not E17G in vesicle study. ► Both E2 and E17G do not compromise CDF formation from CDFDA in hepatocytes. ► CDF accumulation in bile canaliculi is inhibited by E2 or E17G in QTLI. ► Increasing exposure to E2 decreases CDF accumulation in bile canaliculi in QTLI. ► QTLI is feasible to assess Mrp2-based DDI involving drug metabolite in hepatocytes.

  17. Two-photon fluorescence and fluorescence imaging of two styryl heterocyclic dyes combined with DNA.

    Science.gov (United States)

    Gao, Chao; Liu, Shu-yao; Zhang, Xian; Liu, Ying-kai; Qiao, Cong-de; Liu, Zhao-e

    2016-03-05

    Two new styryl heterocyclic two-photon (TP) materials, 4-[4-(N-methyl)styrene]-imidazo [4,5-f][1,10] phenanthroline-benzene iodated salt (probe-1) and 4,4-[4-(N-methyl)styrene]-benzene iodated salt (probe-2) were successfully synthesized and studied as potential fluorescent probes of DNA detection. The linear and nonlinear photophysical properties of two compounds in different solvents were investigated. The absorption, one- and two-photon fluorescent spectra of the free dye and dye-DNA complex were also examined to evaluate their photophysical properties. The binding constants of dye-DNA were obtained according to Scatchard equation with good values. The results showed that two probes could be used as fluorescent DNA probes by two-photon excitation, and TP fluorescent properties of probe-1 are superior to that of probe-2. The fluorescent method date indicated that the mechanisms of dye-DNA complex interaction may be groove binding for probe-1 and electrostatic interaction for probe-2, respectively. The MTT assay experiments showed two probes are low toxicity. Moreover, the TP fluorescence imaging of DNA detection in living cells at 800 nm indicated that the ability to locate in cell nuclei of probe-1 is better than that of probe-2. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Imaging immune response of skin mast cells in vivo with two-photon microscopy

    Science.gov (United States)

    Li, Chunqiang; Pastila, Riikka K.; Lin, Charles P.

    2012-02-01

    Intravital multiphoton microscopy has provided insightful information of the dynamic process of immune cells in vivo. However, the use of exogenous labeling agents limits its applications. There is no method to perform functional imaging of mast cells, a population of innate tissue-resident immune cells. Mast cells are widely recognized as the effector cells in allergy. Recently their roles as immunoregulatory cells in certain innate and adaptive immune responses are being actively investigated. Here we report in vivo mouse skin mast cells imaging with two-photon microscopy using endogenous tryptophan as the fluorophore. We studied the following processes. 1) Mast cells degranulation, the first step in the mast cell activation process in which the granules are released into peripheral tissue to trigger downstream reactions. 2) Mast cell reconstitution, a procedure commonly used to study mast cells functioning by comparing the data from wild type mice, mast cell-deficient mice, and mast-cell deficient mice reconstituted with bone marrow-derived mast cells (BMMCs). Imaging the BMMCs engraftment in tissue reveals the mast cells development and the efficiency of BMMCs reconstitution. We observed the reconstitution process for 6 weeks in the ear skin of mast cell-deficient Kit wsh/ w-sh mice by two-photon imaging. Our finding is the first instance of imaging mast cells in vivo with endogenous contrast.

  19. An automated detection for axonal boutons in vivo two-photon imaging of mouse

    Science.gov (United States)

    Li, Weifu; Zhang, Dandan; Xie, Qiwei; Chen, Xi; Han, Hua

    2017-02-01

    Activity-dependent changes in the synaptic connections of the brain are tightly related to learning and memory. Previous studies have shown that essentially all new synaptic contacts were made by adding new partners to existing synaptic elements. To further explore synaptic dynamics in specific pathways, concurrent imaging of pre and postsynaptic structures in identified connections is required. Consequently, considerable attention has been paid for the automated detection of axonal boutons. Different from most previous methods proposed in vitro data, this paper considers a more practical case in vivo neuron images which can provide real time information and direct observation of the dynamics of a disease process in mouse. Additionally, we present an automated approach for detecting axonal boutons by starting with deconvolving the original images, then thresholding the enhanced images, and reserving the regions fulfilling a series of criteria. Experimental result in vivo two-photon imaging of mouse demonstrates the effectiveness of our proposed method.

  20. Simultaneous morphological and functional imaging of the honeybee's brain by two-photon microscopy

    International Nuclear Information System (INIS)

    Haase, A.

    2011-01-01

    Thanks to its rather simply structured but highly performing brain, the honeybee (Apis mellifera) is an important model for neurobiological studies. Therefore there is a great need for new functional imaging modalities adapted to this species. Herein we give a detailed report on the development and performance of a platform for in vivo functional and morphological imaging of the honeybee's brain, focusing on its primary olfactory centres, the antennal lobes (ALs). The experimental setup consists of a two-photon microscope combined with a synchronized odour stimulus generator. Our imaging platform allows to simultaneously obtain both morphological measurements of the ALs functional units, the glomeruli, and in vivo calcium recording of their neural activity. We were able to record the characteristic glomerular response maps to odour stimuli applied to the bee's antennae. Our approach offers several advantages over the commonly used conventional fluorescence microscopy. Two-photon microscopy provides substantial enhancement in both spatial and temporal resolutions, while minimizing photo damage. Calcium recordings show a more than fourfold improvement in the functional signal with respect to the techniques available up to now. Finally, the extended penetration depth, thanks to the infrared excitation, allows the functional imaging of profound glomeruli which have not been optically accessible up to now.

  1. Hybrid of two-photon microscopy and optical multimodality imaging for multi-scale imaging of small animals

    Science.gov (United States)

    Li, Tianmeng; Hui, Hui; Ma, He; Yang, Xin; Tian, Jie

    2018-02-01

    Non-invasive imaging technologies, such as magnetic resonance imaging (MRI) and optical multimodality imaging methods, are commonly used for diagnosing and supervising the development of inflammatory bowel disease (IBD). These in vivo imaging methods can provide morphology changes information of IBD in macro-scale. However, it is difficult to investigate the intestinal wall in molecular and cellular level. State-of-art light-sheet and two-photon microscopy have the ability to acquire the changes for IBD in micro-scale. The aim of this work is to evaluate the size of the enterocoel and the thickness of colon wall using both MRI for in vivo imaging, and light-sheet and two-photon microscope for in vitro imaging. C57BL/6 mice were received 3.5% Dextran sodium sulfate (DSS) in the drinking water for 5 days to build IBD model. Mice were imaged with MRI on days 0, 6 to observe colitis progression. After MRI imaging, the mice were sacrificed to take colons for tissue clearing. Then, light-sheet and two-photon microscopies are used for in vitro imaging of the cleared samples. The experimental group showed symptoms of bloody stools, sluggishness and weight loss. It showed that the colon wall was thicker while the enterocoel was narrower compare to control group. The more details are observed using light-sheet and two-photon microscope. It is demonstrated that hybrid of MRI in macro-scale and light-sheet and two-photon microscopy in micro-scale imaging is feasible for colon inflammation diagnosing and supervising.

  2. In vivo imaging of cerebral energy metabolism with two-photon fluorescence lifetime microscopy of NADH.

    Science.gov (United States)

    Yaseen, Mohammad A; Sakadžić, Sava; Wu, Weicheng; Becker, Wolfgang; Kasischke, Karl A; Boas, David A

    2013-02-01

    Minimally invasive, specific measurement of cellular energy metabolism is crucial for understanding cerebral pathophysiology. Here, we present high-resolution, in vivo observations of autofluorescence lifetime as a biomarker of cerebral energy metabolism in exposed rat cortices. We describe a customized two-photon imaging system with time correlated single photon counting detection and specialized software for modeling multiple-component fits of fluorescence decay and monitoring their transient behaviors. In vivo cerebral NADH fluorescence suggests the presence of four distinct components, which respond differently to brief periods of anoxia and likely indicate different enzymatic formulations. Individual components show potential as indicators of specific molecular pathways involved in oxidative metabolism.

  3. Simple fibre based dispersion management for two-photon excited fluorescence imaging through an endoscope

    DEFF Research Database (Denmark)

    Dimopoulos, Konstantinos; Marti, Dominik; Andersen, Peter E.

    2018-01-01

    We want to implement two-photon excitation fluorescence microscopy (TPEFM) into endoscopes, since TPEFM can provide relevant biomarkers for cancer staging and grading in hollow organs, endoscopically accessible through natural orifices. However, many obstacles must be overcome, among others...... the delivery of short laser pulses to the distal end of the endoscope. To this avail, we present imaging results using an all-fibre dispersion management scheme in a TPEFM setup. The scheme has been conceived by Jespersen et al. in 20101 and relies on the combination of a single mode fibre with normal...

  4. Imaging hydrogen flames by two-photon, laser-induced fluorescence

    Science.gov (United States)

    Miles, R.; Lempert, W.; Kumar, V.; Diskin, G.

    1991-01-01

    A nonintrusive multicomponent imaging system is developed which can image hydrogen, hot oxygen, and air simultaneously. An Ar-F excimer laser is injection-locked to cover the Q1 two-photon transition in molecular hydrogen which allows the observation of both hot oxygen and cold hydrogen. Rayleigh scattering from the water molecules occurs at the same frequency as the illuminating laser allowing analysis of the air density. Images of ignited and nonignited hydrogen jets are recorded with a high-sensitivity gated video camera. The images permit the analysis of turbulent hydrogen-core jet, the combustion zone, and the surrounding air, and two-dimensional spatial correlations can be made to study the turbulent structure and couplings between different regions of the flow field. The method is of interest to the study of practical combustion systems which employ hydrogen-air diffusion flames.

  5. Two-photon excited autofluorescence imaging of freshly isolated frog retinas.

    Science.gov (United States)

    Lu, Rong-Wen; Li, Yi-Chao; Ye, Tong; Strang, Christianne; Keyser, Kent; Curcio, Christine A; Yao, Xin-Cheng

    2011-06-01

    The purpose of this study was to investigate cellular sources of autofluorescence signals in freshly isolated frog (Rana pipiens) retinas. Equipped with an ultrafast laser, a laser scanning two-photon excitation fluorescence microscope was employed for sub-cellular resolution examination of both sliced and flat-mounted retinas. Two-photon imaging of retinal slices revealed autofluorescence signals over multiple functional layers, including the photoreceptor layer (PRL), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), and ganglion cell layer (GCL). Using flat-mounted retinas, depth-resolved imaging of individual retinal layers further confirmed multiple sources of autofluorescence signals. Cellular structures were clearly observed at the PRL, ONL, INL, and GCL. At the PRL, the autofluorescence was dominantly recorded from the intracellular compartment of the photoreceptors; while mixed intracellular and extracellular autofluorescence signals were observed at the ONL, INL, and GCL. High resolution autofluorescence imaging clearly revealed mosaic organization of rod and cone photoreceptors; and sub-cellular bright autofluorescence spots, which might relate to connecting cilium, was observed in the cone photoreceptors only. Moreover, single-cone and double-cone outer segments could be directly differentiated.

  6. Nanoshells for in vivo imaging using two-photon excitation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Gao Liang; Nammalvar, Vengadesan [Department of Bioengineering, Rice University, Houston, TX 77005 (United States); Vadakkan, Tegy J, E-mail: lg3@rice.edu, E-mail: venkyn@rice.edu [Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030 (United States)

    2011-09-07

    Gold nanoshells have been intensively investigated and applied to various biomedical fields because of their flexible optical tunability and biological compatibility. They hold great potential to serve as luminescent contrast agents excitable with near-infrared (NIR) lasers. In this paper, we describe the development of nanoshells with a peak of plasmon resonance at 800 nm and their subsequent use for in vivo blood vessel imaging using two-photon excitation microscopy at an excitation wavelength of 750 nm. We were able to image single nanoshell particles in blood vessels and generate optical contrast for blood vessel structure using luminescent signals. These results confirm the feasibility of engineering nanoshells with controlled optical properties for single-particle-based in vivo imaging.

  7. Thermally activated delayed fluorescence organic dots for two-photon fluorescence lifetime imaging

    Science.gov (United States)

    He, Tingchao; Ren, Can; Li, Zhuohua; Xiao, Shuyu; Li, Junzi; Lin, Xiaodong; Ye, Chuanxiang; Zhang, Junmin; Guo, Lihong; Hu, Wenbo; Chen, Rui

    2018-05-01

    Autofluorescence is a major challenge in complex tissue imaging when molecules present in the biological tissue compete with the fluorophore. This issue may be resolved by designing organic molecules with long fluorescence lifetimes. The present work reports the two-photon absorption (TPA) properties of a thermally activated delayed fluorescence (TADF) molecule with carbazole as the electron donor and dicyanobenzene as the electron acceptor (i.e., 4CzIPN). The results indicate that 4CzIPN exhibits a moderate TPA cross-section (˜9 × 10-50 cm4 s photon-1), high fluorescence quantum yield, and a long fluorescence lifetime (˜1.47 μs). 4CzIPN was compactly encapsulated into an amphiphilic copolymer via nanoprecipitation to achieve water-soluble organic dots. Interestingly, 4CzIPN organic dots have been utilized in applications involving two-photon fluorescence lifetime imaging (FLIM). Our work aptly demonstrates that TADF molecules are promising candidates of nonlinear optical probes for developing next-generation multiphoton FLIM applications.

  8. Two-photon NADH imaging exposes boundaries of oxygen diffusion in cortical vascular supply regions.

    Science.gov (United States)

    Kasischke, Karl A; Lambert, Elton M; Panepento, Ben; Sun, Anita; Gelbard, Harris A; Burgess, Robert W; Foster, Thomas H; Nedergaard, Maiken

    2011-01-01

    Oxygen transport imposes a possible constraint on the brain's ability to sustain variable metabolic demands, but oxygen diffusion in the cerebral cortex has not yet been observed directly. We show that concurrent two-photon fluorescence imaging of endogenous nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation exposes well-defined boundaries of tissue oxygen diffusion in the mouse cortex. The NADH fluorescence increases rapidly over a narrow, very low pO(2) range with a p(50) of 3.4 ± 0.6 mm Hg, thereby establishing a nearly binary reporter of significant, metabolically limiting hypoxia. The transient cortical tissue boundaries of NADH fluorescence exhibit remarkably delineated geometrical patterns, which define the limits of tissue oxygen diffusion from the cortical microcirculation and bear a striking resemblance to the ideal Krogh tissue cylinder. The visualization of microvessels and their regional contribution to oxygen delivery establishes penetrating arterioles as major oxygen sources in addition to the capillary network and confirms the existence of cortical oxygen fields with steep microregional oxygen gradients. Thus, two-photon NADH imaging can be applied to expose vascular supply regions and to localize functionally relevant microregional cortical hypoxia with micrometer spatial resolution.

  9. Single pulse two photon fluorescence lifetime imaging (SP-FLIM) with MHz pixel rate.

    Science.gov (United States)

    Eibl, Matthias; Karpf, Sebastian; Weng, Daniel; Hakert, Hubertus; Pfeiffer, Tom; Kolb, Jan Philip; Huber, Robert

    2017-07-01

    Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection. Due to the long pulse length in our configuration, more fluorescence photons are generated per pulse, which allows us to derive the lifetime with a single excitation pulse only. In this paper, we show high quality FLIM images acquired at a pixel rate of 1 MHz. This approach is a promising candidate for an easy-to-use and benchtop FLIM system to make this technique available to a wider research community.

  10. Active learning of cortical connectivity from two-photon imaging data

    Science.gov (United States)

    Wang, Ye; Dunson, David; Sapiro, Guillermo; Ringach, Dario

    2018-01-01

    Understanding how groups of neurons interact within a network is a fundamental question in system neuroscience. Instead of passively observing the ongoing activity of a network, we can typically perturb its activity, either by external sensory stimulation or directly via techniques such as two-photon optogenetics. A natural question is how to use such perturbations to identify the connectivity of the network efficiently. Here we introduce a method to infer sparse connectivity graphs from in-vivo, two-photon imaging of population activity in response to external stimuli. A novel aspect of the work is the introduction of a recommended distribution, incrementally learned from the data, to optimally refine the inferred network. Unlike existing system identification techniques, this “active learning” method automatically focuses its attention on key undiscovered areas of the network, instead of targeting global uncertainty indicators like parameter variance. We show how active learning leads to faster inference while, at the same time, provides confidence intervals for the network parameters. We present simulations on artificial small-world networks to validate the methods and apply the method to real data. Analysis of frequency of motifs recovered show that cortical networks are consistent with a small-world topology model. PMID:29718955

  11. Active learning of cortical connectivity from two-photon imaging data.

    Directory of Open Access Journals (Sweden)

    Martín A Bertrán

    Full Text Available Understanding how groups of neurons interact within a network is a fundamental question in system neuroscience. Instead of passively observing the ongoing activity of a network, we can typically perturb its activity, either by external sensory stimulation or directly via techniques such as two-photon optogenetics. A natural question is how to use such perturbations to identify the connectivity of the network efficiently. Here we introduce a method to infer sparse connectivity graphs from in-vivo, two-photon imaging of population activity in response to external stimuli. A novel aspect of the work is the introduction of a recommended distribution, incrementally learned from the data, to optimally refine the inferred network. Unlike existing system identification techniques, this "active learning" method automatically focuses its attention on key undiscovered areas of the network, instead of targeting global uncertainty indicators like parameter variance. We show how active learning leads to faster inference while, at the same time, provides confidence intervals for the network parameters. We present simulations on artificial small-world networks to validate the methods and apply the method to real data. Analysis of frequency of motifs recovered show that cortical networks are consistent with a small-world topology model.

  12. Adapting a compact confocal microscope system to a two-photon excitation fluorescence imaging architecture.

    Science.gov (United States)

    Diaspro, A; Corosu, M; Ramoino, P; Robello, M

    1999-11-01

    Within the framework of a national National Institute of Physics of Matter (INFM) project, we have realised a two-photon excitation (TPE) fluorescence microscope based on a new generation commercial confocal scanning head. The core of the architecture is a mode-locked Ti:Sapphire laser (Tsunami 3960, Spectra Physics Inc., Mountain View, CA) pumped by a high-power (5 W, 532 nm) laser (Millennia V, Spectra Physics Inc.) and an ultracompact confocal scanning head, Nikon PCM2000 (Nikon Instruments, Florence, Italy) using a single-pinhole design. Three-dimensional point-spread function has been measured to define spatial resolution performances. The TPE microscope has been used with a wide range of excitable fluorescent molecules (DAPI, Fura-2, Indo-1, DiOC(6)(3), fluoresceine, Texas red) covering a single photon spectral range from UV to green. An example is reported on 3D imaging of the helical structure of the sperm head of the Octopus Eledone cirrhosa labelled with an UV excitable dye, i.e., DAPI. The system can be easily switched for operating both in conventional and two-photon mode. Copyright 1999 Wiley-Liss, Inc.

  13. Two-photon calcium imaging during fictive navigation in virtual environments

    Directory of Open Access Journals (Sweden)

    Misha Benjamin Ahrens

    2013-06-01

    Full Text Available A full understanding of nervous system function requires recording from large populations of neurons during naturalistic behaviors. Here we enable paralyzed larval zebrafish to fictively navigate two-dimensional virtual environments while we record optically from many neurons with two-photon imaging. Electrical recordings from motor nerves in the tail are decoded into intended forward swims and turns, which are used to update a virtual environment displayed underneath the fish. Several behavioral features - such as turning responses to whole-field motion and dark avoidance - are well-replicated in this virtual setting. We readily observed neuronal populations in the hindbrain with laterally selective responses that correlated with right or left optomotor behavior. We also observed neurons in the habenula, pallium, and midbrain with response properties specific to environmental features. Beyond single-cell correlations, the classification of network activity in such virtual settings promises to reveal principles of brainwide neural dynamics during behavior.

  14. Femtosecond two-photon laser-induced fluorescence of krypton for high-speed flow imaging.

    Science.gov (United States)

    Wang, Yejun; Capps, Cade; Kulatilaka, Waruna D

    2017-02-15

    Ultrashort-pulse (femtosecond-duration) two-photon laser-induced fluorescence (fs-TPLIF) of an inert gas tracer krypton (Kr) is investigated. A detailed spectroscopic study of fluorescence channels followed by the 5p'←←4p excitation of Kr at 204.1 nm is reported. The experimental line positions in the 750-840 nm emission region agree well with the NIST Atomic Spectra Database. The present work provides an accurate listing of relative line strengths in this spectral region. In the range of laser pulse energies investigated, a quadratic dependence was observed between the Kr-TPLIF signal and the laser pulse energy. The single-laser-shot 2D TPLIF images recorded in an unsteady jet demonstrate the potential of using fs excitation at 204.1 nm for mixing and flow diagnostic studies using Kr as an inert gas tracer.

  15. Intravital Confocal and Two-photon Imaging of Dual-color Cells and Extracellular Matrix Mimics

    Science.gov (United States)

    Bal, Ufuk; Andresen, Volker; Baggett, Brenda; Utzinger, Urs

    2013-01-01

    To optimize imaging of cells in three dimensional culture we studied confocal backscattering, Second Harmonic Generation (SHG) and autofluorescence as source of contrast in extracellular matrix (ECM) mimics and evaluated the attenuation as well as bleaching of endogenous cellular fluorescence signals. All common ECM mimics exhibit contrast observable with confocal reflectance microscopy. SHG imaging on collagen I based hydrogels provides high contrast and good optical penetration depth. Agarose is a useful embedding medium because it allows for large optical penetration and exhibits minimal autofluorescence while still providing good reflectance to detect voids in the embedding medium. We labeled breast cancer cells’ outline with DsRed2 and nucleus with eGFP. DsRed2 can be excited with confocal imaging at 568nm, and with two photon excitation (TPE) in the red and longer NIR. eGFP was excited at 488nm for confocal and in the NIR for TPE. While there is small difference in the bleaching rate for eGFP between confocal and TPE we observed significant difference for DsRed2 where bleaching is strongest during TPE in the red wavelengths and smallest during confocal imaging. After a few hundred microns depth in a collagen I hydrogel, TPE fluorescence becomes twice as strong compared to confocal imaging. PMID:23380006

  16. Time gated fluorescence lifetime imaging and micro-volume spectroscopy using two-photon excitation

    NARCIS (Netherlands)

    Sytsma, J.; Vroom, J.M.; de Grauw, C.J.; Gerritsen, H.C.

    A scanning microscope utilizing two-photon excitation in combination with fluorescence lifetime contrast is presented. The microscope makes use of a tunable femtosecond titanium:sapphire laser enabling the two-photon excitation of a broad range of fluorescent molecules, including UV probes.

  17. Automatic quantification of mitochondrial fragmentation from two-photon microscope images of mouse brain tissue.

    Science.gov (United States)

    Lihavainen, E; Kislin, M; Toptunov, D; Khiroug, L; Ribeiro, A S

    2015-12-01

    The morphology of mitochondria can inform about their functional state and, thus, about cell vitality. For example, fragmentation of the mitochondrial network is associated with many diseases. Recent advances in neuronal imaging have enabled the observation of mitochondria in live brains for long periods of time, enabling the study of their dynamics in animal models of diseases. To aid these studies, we developed an automatic method, based on supervised learning, for quantifying the degree of mitochondrial fragmentation in tissue images acquired via two-photon microscopy from transgenic mice, which exclusively express Enhanced cyan fluorescent protein (ECFP) under Thy1 promoter, targeted to the mitochondrial matrix in subpopulations of neurons. We tested the method on images prior to and after cardiac arrest, and found it to be sensitive to significant changes in mitochondrial morphology because of the arrest. We conclude that the method is useful in detecting morphological abnormalities in mitochondria and, likely, in other subcellular structures as well. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

  18. Imaging-guided two-photon excitation-emission-matrix measurements of human skin tissues

    Science.gov (United States)

    Yu, Yingqiu; Lee, Anthony M. D.; Wang, Hequn; Tang, Shuo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2012-07-01

    There are increased interests on using multiphoton imaging and spectroscopy for skin tissue characterization and diagnosis. However, most studies have been done with just a few excitation wavelengths. Our objective is to perform a systematic study of the two-photon fluorescence (TPF) properties of skin fluorophores, normal skin, and diseased skin tissues. A nonlinear excitation-emission-matrix (EEM) spectroscopy system with multiphoton imaging guidance was constructed. A tunable femtosecond laser was used to vary excitation wavelengths from 730 to 920 nm for EEM data acquisition. EEM measurements were performed on excised fresh normal skin tissues, seborrheic keratosis tissue samples, and skin fluorophores including: NADH, FAD, keratin, melanin, collagen, and elastin. We found that in the stratum corneum and upper epidermis of normal skin, the cells have large sizes and the TPF originates from keratin. In the lower epidermis, cells are smaller and TPF is dominated by NADH contributions. In the dermis, TPF is dominated by elastin components. The depth resolved EEM measurements also demonstrated that keratin structure has intruded into the middle sublayers of the epidermal part of the seborrheic keratosis lesion. These results suggest that the imaging guided TPF EEM spectroscopy provides useful information for the development of multiphoton clinical devices for skin disease diagnosis.

  19. Measurement of drug-target engagement in live cells by two-photon fluorescence anisotropy imaging.

    Science.gov (United States)

    Vinegoni, Claudio; Fumene Feruglio, Paolo; Brand, Christian; Lee, Sungon; Nibbs, Antoinette E; Stapleton, Shawn; Shah, Sunil; Gryczynski, Ignacy; Reiner, Thomas; Mazitschek, Ralph; Weissleder, Ralph

    2017-07-01

    The ability to directly image and quantify drug-target engagement and drug distribution with subcellular resolution in live cells and whole organisms is a prerequisite to establishing accurate models of the kinetics and dynamics of drug action. Such methods would thus have far-reaching applications in drug development and molecular pharmacology. We recently presented one such technique based on fluorescence anisotropy, a spectroscopic method based on polarization light analysis and capable of measuring the binding interaction between molecules. Our technique allows the direct characterization of target engagement of fluorescently labeled drugs, using fluorophores with a fluorescence lifetime larger than the rotational correlation of the bound complex. Here we describe an optimized protocol for simultaneous dual-channel two-photon fluorescence anisotropy microscopy acquisition to perform drug-target measurements. We also provide the necessary software to implement stream processing to visualize images and to calculate quantitative parameters. The assembly and characterization part of the protocol can be implemented in 1 d. Sample preparation, characterization and imaging of drug binding can be completed in 2 d. Although currently adapted to an Olympus FV1000MPE microscope, the protocol can be extended to other commercial or custom-built microscopes.

  20. Fluorenyl benzothiadiazole and benzoselenadiazole near-IR fluorescent probes for two-photon fluorescence imaging (Conference Presentation)

    Science.gov (United States)

    Belfield, Kevin D.; Yao, Sheng; Kim, Bosung; Yue, Xiling

    2016-03-01

    Imaging biological samples with two-photon fluorescence (2PF) microscopy has the unique advantage of resulting high contrast 3D resolution subcellular image that can reach up to several millimeters depth. 2PF probes that absorb and emit at near IR region need to be developed. Two-photon excitation (2PE) wavelengths are less concerned as 2PE uses wavelengths doubles the absorption wavelength of the probe, which means 2PE wavelengths for probes even with absorption at visible wavelength will fall into NIR region. Therefore, probes that fluoresce at near IR region with high quantum yields are needed. A series of dyes based on 5-thienyl-2, 1, 3-benzothiadiazole and 5-thienyl-2, 1, 3-benzoselenadiazole core were synthesized as near infrared two-photon fluorophores. Fluorescence maxima wavelengths as long as 714 nm and fluorescence quantum yields as high as 0.67 were achieved. The fluorescence quantum yields of the dyes were nearly constant, regardless of solvents polarity. These diazoles exhibited large Stokes shift (GM), and high two-photon fluorescence figure of merit (FM , 1.04×10-2 GM). Cells incubated on a 3D scaffold with one of the new probes (encapsulated in Pluronic micelles) exhibited bright fluorescence, enabling 3D two-photon fluorescence imaging to a depth of 100 µm.

  1. Spatiotemporal dynamics of rhythmic spinal interneurons measured with two-photon calcium imaging and coherence analysis.

    Science.gov (United States)

    Kwan, Alex C; Dietz, Shelby B; Zhong, Guisheng; Harris-Warrick, Ronald M; Webb, Watt W

    2010-12-01

    In rhythmic neural circuits, a neuron often fires action potentials with a constant phase to the rhythm, a timing relationship that can be functionally significant. To characterize these phase preferences in a large-scale, cell type-specific manner, we adapted multitaper coherence analysis for two-photon calcium imaging. Analysis of simulated data showed that coherence is a simple and robust measure of rhythmicity for calcium imaging data. When applied to the neonatal mouse hindlimb spinal locomotor network, the phase relationships between peak activity of >1,000 ventral spinal interneurons and motor output were characterized. Most interneurons showed rhythmic activity that was coherent and in phase with the ipsilateral motor output during fictive locomotion. The phase distributions of two genetically identified classes of interneurons were distinct from the ensemble population and from each other. There was no obvious spatial clustering of interneurons with similar phase preferences. Together, these results suggest that cell type, not neighboring neuron activity, is a better indicator of an interneuron's response during fictive locomotion. The ability to measure the phase preferences of many neurons with cell type and spatial information should be widely applicable for studying other rhythmic neural circuits.

  2. High-resolution, label-free two-photon imaging of diseased human corneas

    Science.gov (United States)

    Batista, Ana; Breunig, Hans Georg; König, Aisada; Schindele, Andreas; Hager, Tobias; Seitz, Berthold; König, Karsten

    2018-03-01

    The diagnosis of corneal diseases may be improved by monitoring the metabolism of cells and the structural organization of the stroma using two-photon imaging (TPI). We used TPI to assess the differences between nonpathological (NP) human corneas and corneas diagnosed with either keratoconus, Acanthamoeba keratitis, or stromal corneal scars. Images were acquired using a custom-built five-dimensional laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed excitation laser and a 16-channel photomultiplier tube detector in combination with a time-correlated single photon counting module. Morphological alterations of epithelial cells were observed for all pathologies. Moreover, diseased corneas showed alterations to the cells' metabolism that were revealed using the NAD(P)H free to protein-bound ratios. The mean autofluorescence lifetime of the stroma and the organization of the collagen fibers were also significantly altered due to the pathologies. We demonstrate that TPI can be used to distinguish between NP and diseased human corneas, based not only on alterations of the cells' morphology, which can also be evaluated using current clinical devices, but on additional morphological and functional features such as the organization of the stroma and the cells' metabolism. Therefore, TPI could become an efficient tool for diagnosing corneal diseases and better understanding the biological processes of the diseases.

  3. Linking Neurons to Network Function and Behavior by Two-Photon Holographic Optogenetics and Volumetric Imaging.

    Science.gov (United States)

    Dal Maschio, Marco; Donovan, Joseph C; Helmbrecht, Thomas O; Baier, Herwig

    2017-05-17

    We introduce a flexible method for high-resolution interrogation of circuit function, which combines simultaneous 3D two-photon stimulation of multiple targeted neurons, volumetric functional imaging, and quantitative behavioral tracking. This integrated approach was applied to dissect how an ensemble of premotor neurons in the larval zebrafish brain drives a basic motor program, the bending of the tail. We developed an iterative photostimulation strategy to identify minimal subsets of channelrhodopsin (ChR2)-expressing neurons that are sufficient to initiate tail movements. At the same time, the induced network activity was recorded by multiplane GCaMP6 imaging across the brain. From this dataset, we computationally identified activity patterns associated with distinct components of the elicited behavior and characterized the contributions of individual neurons. Using photoactivatable GFP (paGFP), we extended our protocol to visualize single functionally identified neurons and reconstruct their morphologies. Together, this toolkit enables linking behavior to circuit activity with unprecedented resolution. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Plasmon-resonant nanorods as multimodal agents for two-photon luminescent imaging and photothermal therapy

    Science.gov (United States)

    Huff, Terry B.; Hansen, Matthew N.; Tong, Ling; Zhao, Yan; Wang, Haifeng; Zweifel, Daniel A.; Cheng, Ji-Xin; Wei, Alexander

    2007-02-01

    Plasmon-resonant gold nanorods have outstanding potential as multifunctional agents for image-guided therapies. Nanorods have large absorption cross sections at near-infrared (NIR) frequencies, and produce two-photon luminescence (TPL) when excited by fs-pulsed laser irradiation. The TPL signals can be detected with single-particle sensitivity, enabling nanorods to be imaged in vivo while passing through blood vessels at subpicomolar concentrations. Furthermore, cells labeled with nanorods become highly susceptible to photothermal damage when irradiated at plasmon resonance, often resulting in a dramatic blebbing of the cell membrane. However, the straightforward application of gold nanorods for cell-specific labeling is obstructed by the presence of CTAB, a cationic surfactant carried over from nanorod synthesis which also promotes their nonspecific uptake into cells. Careful exchange and replacement of CTAB can be achieved by introducing oligoethyleneglycol (OEG) units capable of chemisorption onto nanorod surfaces by in situ dithiocarbamate formation, a novel method of surface functionalization. Nanorods with a dense coating of methyl-terminated OEG chains are shielded from nonspecific cell uptake, whereas nanorods functionalized with folate-terminated OEG chains accumulate on the surface of tumor cells overexpressing their cognate receptor, with subsequent delivery of photoinduced cell damage at low laser fluence.

  5. Identification and two-photon imaging of oligodendrocyte in CA1 region of hippocampal slices

    International Nuclear Information System (INIS)

    Zhou Wei; Ge Wooping; Zeng Shaoqun; Duan Shumin; Luo Qingming

    2007-01-01

    Oligodendrocyte (OL) plays a critical role in myelination and axon maintenance in central nervous system. Recent studies show that OL can also express NMDA receptors in development and pathological situations in white matter. There is still lack of studies about OL properties and function in gray matter of brain. Here we reported that some glial cells in CA1 region of rat hippocampal slices (P15-23) had distinct electrophysiological characteristics from the other glia cells in this region, while they displayed uniform properties with OL from white matter in previous report; therefore, they were considered as OL in hippocampus. By loading dye in recording pipette and imaging with two-photon laser scanning microscopy, we acquired the high spatial resolution, three-dimension images of these special cells in live slices. The OL in hippocampus shows a complex process-bearing shape and the distribution of several processes is parallel to Schaffer fiber in CA1 region. When stimulating Schaffer fiber, OL displays a long duration depolarization mediated by inward rectifier potassium channel. This suggested that the OL in CA1 region could sense the neuronal activity and contribute to potassium clearance

  6. Mannose-functionalized porous silica-coated magnetic nanoparticles for two-photon imaging or PDT of cancer cells

    International Nuclear Information System (INIS)

    Perrier, Marine; Gary-Bobo, Magali; Lartigue, Lenaïc; Brevet, David; Morère, Alain; Garcia, Marcel; Maillard, Philippe; Raehm, Laurence; Guari, Yannick; Larionova, Joulia; Durand, Jean-Olivier; Mongin, Olivier; Blanchard-Desce, Mireille

    2013-01-01

    An original fluorophore engineered for two-photon excitation or a porphyrin derivative were entrapped in the silica shell of magnetic porous silica nanoparticles during the synthesis of the silica moiety without damaging the structure of the organic part. The mild conditions involved allowed obtaining microporous or mesoporous silica magnetic nanoparticles, respectively. Mannose was grafted on the surface of the nanoparticles to target MCF-7 breast cancer cells. The studies of magnetic properties of these hybrid nanoparticles show that they present a blocking temperature at 190 K. The nano-objects designed with the two-photon fluorophore were efficient for two-photon imaging of MCF-7 cancer cells, whereas the nano-objects with the photosensitizer efficiently killed cancer cells. The presence of the mannose moiety was demonstrated to improve both imaging and therapy properties.

  7. Mannose-functionalized porous silica-coated magnetic nanoparticles for two-photon imaging or PDT of cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Perrier, Marine [UMR 5253 CNRS-UM2-ENSCM-UM1, Institut Charles Gerhardt Montpellier (France); Gary-Bobo, Magali [Faculte de Pharmacie, Universite Montpellier 1, Universite Montpellier 2, Institut des Biomolecules Max Mousseron UMR 5247 CNRS (France); Lartigue, Lenaiec; Brevet, David [UMR 5253 CNRS-UM2-ENSCM-UM1, Institut Charles Gerhardt Montpellier (France); Morere, Alain; Garcia, Marcel [Faculte de Pharmacie, Universite Montpellier 1, Universite Montpellier 2, Institut des Biomolecules Max Mousseron UMR 5247 CNRS (France); Maillard, Philippe [Universite Paris-Sud, UMR 176 CNRS, Institut Curie (France); Raehm, Laurence; Guari, Yannick, E-mail: yannick.guari@um2.fr; Larionova, Joulia; Durand, Jean-Olivier, E-mail: durand@univ-montp2.fr [UMR 5253 CNRS-UM2-ENSCM-UM1, Institut Charles Gerhardt Montpellier (France); Mongin, Olivier [Universite de Rennes 1, Institut des Sciences Chimiques de Rennes, CNRS UMR 6226 (France); Blanchard-Desce, Mireille [Universite Bordeaux, Institut des Sciences Moleculaires, UMR CNRS 5255 (France)

    2013-05-15

    An original fluorophore engineered for two-photon excitation or a porphyrin derivative were entrapped in the silica shell of magnetic porous silica nanoparticles during the synthesis of the silica moiety without damaging the structure of the organic part. The mild conditions involved allowed obtaining microporous or mesoporous silica magnetic nanoparticles, respectively. Mannose was grafted on the surface of the nanoparticles to target MCF-7 breast cancer cells. The studies of magnetic properties of these hybrid nanoparticles show that they present a blocking temperature at 190 K. The nano-objects designed with the two-photon fluorophore were efficient for two-photon imaging of MCF-7 cancer cells, whereas the nano-objects with the photosensitizer efficiently killed cancer cells. The presence of the mannose moiety was demonstrated to improve both imaging and therapy properties.

  8. Capturing change: the duality of time-lapse imagery to acquire data and depict ecological dynamics

    Science.gov (United States)

    Brinley Buckley, Emma M.; Allen, Craig R.; Forsberg, Michael; Farrell, Michael; Caven, Andrew J.

    2017-01-01

    We investigate the scientific and communicative value of time-lapse imagery by exploring applications for data collection and visualization. Time-lapse imagery has a myriad of possible applications to study and depict ecosystems and can operate at unique temporal and spatial scales to bridge the gap between large-scale satellite imagery projects and observational field research. Time-lapse data sequences, linking time-lapse imagery with data visualization, have the ability to make data come alive for a wider audience by connecting abstract numbers to images that root data in time and place. Utilizing imagery from the Platte Basin Timelapse Project, water inundation and vegetation phenology metrics are quantified via image analysis and then paired with passive monitoring data, including streamflow and water chemistry. Dynamic and interactive time-lapse data sequences elucidate the visible and invisible ecological dynamics of a significantly altered yet internationally important river system in central Nebraska.

  9. Capturing change: the duality of time-lapse imagery to acquire data and depict ecological dynamics

    Directory of Open Access Journals (Sweden)

    Emma M. Brinley Buckley

    2017-09-01

    Full Text Available We investigate the scientific and communicative value of time-lapse imagery by exploring applications for data collection and visualization. Time-lapse imagery has a myriad of possible applications to study and depict ecosystems and can operate at unique temporal and spatial scales to bridge the gap between large-scale satellite imagery projects and observational field research. Time-lapse data sequences, linking time-lapse imagery with data visualization, have the ability to make data come alive for a wider audience by connecting abstract numbers to images that root data in time and place. Utilizing imagery from the Platte Basin Timelapse Project, water inundation and vegetation phenology metrics are quantified via image analysis and then paired with passive monitoring data, including streamflow and water chemistry. Dynamic and interactive time-lapse data sequences elucidate the visible and invisible ecological dynamics of a significantly altered yet internationally important river system in central Nebraska.

  10. Two-Photon Probes for Lysosomes and Mitochondria: Simultaneous Detection of Lysosomes and Mitochondria in Live Tissues by Dual-Color Two-Photon Microscopy Imaging.

    Science.gov (United States)

    Lim, Chang Su; Hong, Seung Taek; Ryu, Seong Shick; Kang, Dong Eun; Cho, Bong Rae

    2015-10-01

    Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Review: two-photon scanning systems for clinical high resolution in vivo tissue imaging

    Science.gov (United States)

    König, K.; Müller, J.; Höfer, M.; Müller, C.; Weinigel, M.; Bückle, R.; Elsner, P.; Kaatz, M.; Messerschmidt, B.

    2008-02-01

    The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as

  12. Two-Photon Functional Imaging of the Auditory Cortex in Behaving Mice: From Neural Networks to Single Spines

    Directory of Open Access Journals (Sweden)

    Ruijie Li

    2018-04-01

    Full Text Available In vivo two-photon Ca2+ imaging is a powerful tool for recording neuronal activities during perceptual tasks and has been increasingly applied to behaving animals for acute or chronic experiments. However, the auditory cortex is not easily accessible to imaging because of the abundant temporal muscles, arteries around the ears and their lateral locations. Here, we report a protocol for two-photon Ca2+ imaging in the auditory cortex of head-fixed behaving mice. By using a custom-made head fixation apparatus and a head-rotated fixation procedure, we achieved two-photon imaging and in combination with targeted cell-attached recordings of auditory cortical neurons in behaving mice. Using synthetic Ca2+ indicators, we recorded the Ca2+ transients at multiple scales, including neuronal populations, single neurons, dendrites and single spines, in auditory cortex during behavior. Furthermore, using genetically encoded Ca2+ indicators (GECIs, we monitored the neuronal dynamics over days throughout the process of associative learning. Therefore, we achieved two-photon functional imaging at multiple scales in auditory cortex of behaving mice, which extends the tool box for investigating the neural basis of audition-related behaviors.

  13. Detection of axonal synapses in 3D two-photon images.

    Directory of Open Access Journals (Sweden)

    Cher Bass

    Full Text Available Studies of structural plasticity in the brain often require the detection and analysis of axonal synapses (boutons. To date, bouton detection has been largely manual or semi-automated, relying on a step that traces the axons before detection the boutons. If tracing the axon fails, the accuracy of bouton detection is compromised. In this paper, we propose a new algorithm that does not require tracing the axon to detect axonal boutons in 3D two-photon images taken from the mouse cortex. To find the most appropriate techniques for this task, we compared several well-known algorithms for interest point detection and feature descriptor generation. The final algorithm proposed has the following main steps: (1 a Laplacian of Gaussian (LoG based feature enhancement module to accentuate the appearance of boutons; (2 a Speeded Up Robust Features (SURF interest point detector to find candidate locations for feature extraction; (3 non-maximum suppression to eliminate candidates that were detected more than once in the same local region; (4 generation of feature descriptors based on Gabor filters; (5 a Support Vector Machine (SVM classifier, trained on features from labelled data, and was used to distinguish between bouton and non-bouton candidates. We found that our method achieved a Recall of 95%, Precision of 76%, and F1 score of 84% within a new dataset that we make available for accessing bouton detection. On average, Recall and F1 score were significantly better than the current state-of-the-art method, while Precision was not significantly different. In conclusion, in this article we demonstrate that our approach, which is independent of axon tracing, can detect boutons to a high level of accuracy, and improves on the detection performance of existing approaches. The data and code (with an easy to use GUI used in this article are available from open source repositories.

  14. Definition of "banner clouds" based on time lapse movies

    Directory of Open Access Journals (Sweden)

    J. H. Schween

    2007-01-01

    Full Text Available Banner clouds appear on the leeward side of a mountain and resemble a banner or a flag. This article provides a comprehensive definition of "banner clouds". It is based primarily on an extensive collection of time lapse movies, but previous attempts at an explanation of this phenomenon are also taken into account. The following ingredients are considered essential: the cloud must be attached to the mountain but not appear on the windward side; the cloud must originate from condensation of water vapour contained in the air (rather than consist of blowing snow; the cloud must be persistent; and the cloud must not be of convective nature. The definition is illustrated and discussed with the help of still images and time lapse movies taken at Mount Zugspitze in the Bavarian Alps.

  15. Phosphorus ligand imaging with two-photon fluorescence spectroscopy: towards rational catalyst immobilization

    NARCIS (Netherlands)

    Marras, F.; Kluwer, A.M.; Siekierzycka, J.R.; Vozza, A.; Brouwer, A.M.; Reek, J.N.H.

    2010-01-01

    Spotless catalysts: Ligand immobilization was studied by two-photon fluorescence microscopy with a fluorescent nixantphos ligand as probe (see picture). In the immobilization process ligand aggregates form in solution and are deposited on the support, where they appear as bright spots in

  16. Two-photon imaging of field enhancement by groups of gold nanostrip antennas

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Sondergaard, Thomas

    2009-01-01

    Resonant field enhancement by groups of 16 nm thin gold nanostrip antennas consisting of four strips (widths of 70, 100, and 130 nm) and fixed gap (50, 100, 150, or 200 nm) between them and positioned on a quartz substrate is investigated by reflection spectroscopy and two-photon photoluminescenc...

  17. Label-free imaging immune cells and collagen in atherosclerosis with two-photon and second harmonic generation microscopy

    Directory of Open Access Journals (Sweden)

    Chunqiang Li

    2016-01-01

    Full Text Available Atherosclerosis has been recognized as a chronic inflammation disease, in which many types of cells participate in this process, including lymphocytes, macrophages, dendritic cells (DCs, mast cells, vascular smooth muscle cells (SMCs. Developments in imaging technology provide the capability to observe cellular and tissue components and their interactions. The knowledge of the functions of immune cells and their interactions with other cell and tissue components will facilitate our discovery of biomarkers in atherosclerosis and prediction of the risk factor of rupture-prone plaques. Nonlinear optical microscopy based on two-photon excited autofluorescence and second harmonic generation (SHG were developed to image mast cells, SMCs and collagen in plaque ex vivo using endogenous optical signals. Mast cells were imaged with two-photon tryptophan autofluorescence, SMCs were imaged with two-photon NADH autofluorescence, and collagen were imaged with SHG. This development paves the way for further study of mast cell degranulation, and the effects of mast cell derived mediators such as induced synthesis and activation of matrix metalloproteinases (MMPs which participate in the degradation of collagen.

  18. Water-Soluble Triarylborane Chromophores for One- and Two-Photon Excited Fluorescence Imaging of Mitochondria in Cells.

    Science.gov (United States)

    Griesbeck, Stefanie; Zhang, Zuolun; Gutmann, Marcus; Lühmann, Tessa; Edkins, Robert M; Clermont, Guillaume; Lazar, Adina N; Haehnel, Martin; Edkins, Katharina; Eichhorn, Antonius; Blanchard-Desce, Mireille; Meinel, Lorenz; Marder, Todd B

    2016-10-04

    Three water-soluble tetracationic quadrupolar chromophores comprising two three-coordinate boron π-acceptor groups bridged by thiophene-containing moieties were synthesised for biological imaging applications. Compound 3 containing the bulkier 5-(3,5-Me2 C6 H2 )-2,2'-(C4 H2 S)2 -5'-(3,5-Me2 C6 H2 ) bridge is stable over a long period of time, exhibits a high fluorescence quantum yield and strong one- and two-photon absorption (TPA), and has a TPA cross section of 268 GM at 800 nm in water. Confocal laser scanning fluorescence microscopy studies in live cells indicated localisation of the chromophore at the mitochondria; moreover, cytotoxicity measurements proved biocompatibility. Thus, chromophore 3 has excellent potential for one- and two-photon-excited fluorescence imaging of mitochondrial function in cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Imaging of Fluoride Ion in Living Cells and Tissues with a Two-Photon Ratiometric Fluorescence Probe

    Directory of Open Access Journals (Sweden)

    Xinyue Zhu

    2015-01-01

    Full Text Available A reaction-based two-photon (TP ratiometric fluorescence probe Z2 has been developed and successfully applied to detect and image fluoride ion in living cells and tissues. The Z2 probe was designed designed to utilize an ICT mechanism between n-butylnaphthalimide as a fluorophore and tert-butyldiphenylsilane (TBDPS as a response group. Upon addition of fluoride ion, the Si-O bond in the Z2 would be cleaved, and then a stronger electron-donating group was released. The fluorescent changes at 450 and 540 nm, respectively, made it possible to achieve ratiometric fluorescence detection. The results indicated that the Z2 could ratiometrically detect and image fluoride ion in living cells and tissues in a depth of 250 μm by two-photon microscopy (TPM.

  20. Time-Lapse Measurement of Wellbore Integrity

    Science.gov (United States)

    Duguid, A.

    2017-12-01

    Well integrity is becoming more important as wells are used longer or repurposed. For CO2, shale gas, and other projects it has become apparent that wells represent the most likely unintended migration pathway for fluids out of the reservoir. Comprehensive logging programs have been employed to determine the condition of legacy wells in North America. These studies provide examples of assessment technologies. Logging programs have included pulsed neutron logging, ultrasonic well mapping, and cement bond logging. While these studies provide examples of what can be measured, they have only conducted a single round of logging and cannot show if the well has changed over time. Recent experience with time-lapse logging of three monitoring wells at a US Department of Energy sponsored CO2 project has shown the full value of similar tools. Time-lapse logging has shown that well integrity changes over time can be identified. It has also shown that the inclusion of and location of monitoring technologies in the well and the choice of construction materials must be carefully considered. Two of the wells were approximately eight years old at the time of study; they were constructed with steel and fiberglass casing sections and had lines on the outside of the casing running to the surface. The third well was 68 years old when it was studied and was originally constructed as a production well. Repeat logs were collected six or eight years after initial logging. Time-lapse logging showed the evolution of the wells. The results identified locations where cement degraded over time and locations that showed little change. The ultrasonic well maps show clearly that the lines used to connect the monitoring technology to the surface are visible and have a local effect on cement isolation. Testing and sampling was conducted along with logging. It provided insight into changes identified in the time-lapse log results. Point permeability testing was used to provide an in-situ point

  1. Time lapse microscopy observation of cellular structural changes and image analysis of drug treated cancer cells to characterize the cellular heterogeneity.

    Science.gov (United States)

    Vaiyapuri, Periasamy S; Ali, Alshatwi A; Mohammad, Akbarsha A; Kandhavelu, Jeyalakshmi; Kandhavelu, Meenakshisundaram

    2015-01-01

    The effect of Calotropis gigantea latex (CGLX) on human mammary carcinoma cells is not well established. We present the results of this drug activity at total population and single cell level. CGLX inhibited the growth of MCF7 cancer cells at lower IC50 concentration (17 µL/mL). Microscopy of IC50 drug treated cells at 24 hr confirming the appearance of morphological characteristics of apoptotic and necrotic cells, associated with 70% of DNA damage. FACS analysis confirmed that, 10 and 20% of the disruption of cellular mitochondrial nature by at 24 and 48 h, respectively. Microscopic image analysis of total population level proved that MMP changes were statistically significant with P values. The cell to cell variation was confirmed by functional heterogeneity analysis which proves that CGLX was able to induce the apoptosis without the contribution of mitochondria. We conclude that CGLX inhibits cell proliferation, survival, and heterogeneity of pathways in human mammary carcinoma cells. © 2014 Wiley Periodicals, Inc.

  2. Human female meiosis revised: new insights into the mechanisms of chromosome segregation and aneuploidies from advanced genomics and time-lapse imaging.

    Science.gov (United States)

    Capalbo, Antonio; Hoffmann, Eva R; Cimadomo, Danilo; Ubaldi, Filippo Maria; Rienzi, Laura

    2017-11-01

    The unbalanced transmission of chromosomes in human gametes and early preimplantation embryos causes aneuploidy, which is a major cause of infertility and pregnancy failure. A baseline of 20% of human oocytes are estimated to be aneuploid and this increases exponentially from 30 to 35 years, reaching on average 80% by 42 years. As a result, reproductive senescence in human females is predominantly determined by the accelerated decline in genetic quality of oocytes from 30 years of age. Understanding mechanisms of chromosome segregation and aneuploidies in the female germline is a crucial step towards the development of new diagnostic approaches and, possibly, for the development of therapeutic targets and molecules. Here, we have reviewed emerging mechanisms that may drive human aneuploidy, in particular the maternal age effect. We conducted a systematic search in PubMed Central of the primary literature from 1990 through 2016 following the PRISMA guidelines, using MeSH terms related to human aneuploidy. For model organism research, we conducted a literature review based on references in human oocytes manuscripts and general reviews related to chromosome segregation in meiosis and mitosis. Advances in genomic and imaging technologies are allowing unprecedented insight into chromosome segregation in human oocytes. This includes the identification of a novel chromosome segregation error, termed reverse segregation, as well as sister kinetochore configurations that were not predicted based on murine models. Elucidation of mechanisms that result in errors in chromosome segregation in meiosis may lead to therapeutic developments that could improve reproductive outcomes by reducing aneuploidy. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com

  3. Time-lapse integrated geophysical imaging of magmatic injections and fluid-induced fracturing causing Campi Flegrei 1983-84 Unrest

    Science.gov (United States)

    De Siena, Luca; Crescentini, Luca; Amoruso, Antonella; Del Pezzo, Edoardo; Castellano, Mario

    2016-04-01

    Geophysical precursors measured during Unrest episodes are a primary source of geophysical information to forecast eruptions at the largest and most potentially destructive volcanic calderas. Despite their importance and uniqueness, these precursors are also considered difficult to interpret and unrepresentative of larger eruptive events. Here, we show how novel geophysical imaging and monitoring techniques are instead able to represent the dynamic evolution of magmatic- and fluid-induced fracturing during the largest period of Unrest at Campi Flegrei caldera, Italy (1983-1984). The time-dependent patterns drawn by microseismic locations and deformation, once integrated by 3D attenuation tomography and absorption/scattering mapping, model injections of magma- and fluid-related materials in the form of spatially punctual microseismic bursts at a depth of 3.5 km, west and offshore the city of Pozzuoli. The shallowest four kilometres of the crust work as a deformation-based dipolar system before and after each microseismic shock. Seismicity and deformation contemporaneously focus on the point of injection; patterns then progressively crack the medium directed towards the second focus, a region at depths 1-1.5 km south of Solfatara. A single high-absorption and high-scattering aseismic anomaly marks zones of fluid storage overlying the first dipolar centre. These results provide the first direct geophysical signature of the processes of aseismic fluid release at the top of the basaltic basement, producing pozzolanic activity and recently observed via rock-physics and well-rock experiments. The microseismicity caused by fluids and gasses rises to surface via high-absorption north-east rising paths connecting the two dipolar centres, finally beingq being generally expelled from the maar diatreme Solfatara structure. Geophysical precursors during Unrest depict how volcanic stress was released at the Campi Flegrei caldera during its period of highest recorded seismicity

  4. 3D imaging of intrinsic crystalline defects in zinc oxide by spectrally resolved two-photon fluorescence microscopy

    Science.gov (United States)

    Al-Tabich, A.; Inami, W.; Kawata, Y.; Jablonski, R.; Worasawat, S.; Mimura, H.

    2017-05-01

    We present a method for three-dimensional intrinsic defect imaging in zinc oxide (ZnO) by spectrally resolved two-photon fluorescence microscopy, based on the previously presented method of observing a photoluminescence distribution in wide-gap semiconductor crystals [Noor et al., Appl. Phys. Lett. 92(16), 161106 (2008)]. A tightly focused light beam radiated by a titanium-sapphire laser is used to obtain a two-photon excitation of selected area of the ZnO sample. Photoluminescence intensity of a specific spectral range is then selected by optical band pass filters and measured by a photomultiplier tube. Reconstruction of the specimen image is done by scanning the volume of interest by a piezoelectric positioning stage and measuring the spectrally resolved photoluminescence intensity at each point. The method has been proved to be effective at locating intrinsic defects of the ZnO crystalline structure in the volume of the crystal. The method was compared with other defect imaging and 3D imaging techniques like scanning tunneling microscopy and confocal microscopy. In both cases, our method shows superior penetration abilities and, as the only method, allows location of the defects of the chosen type in 3D. In this paper, we present the results of oxygen vacancies and zinc antisites imaging in ZnO nanorods.

  5. Time-lapse seismic attribute analysis for a water-flooded reservoir

    International Nuclear Information System (INIS)

    Jin, Long; Sen, M K; Stoffa, P L; Seif, R K

    2008-01-01

    One of the goals of time-lapse seismic monitoring is the direct detection of the fluid front and two-phase contact area. However, several factors affect the quality of time-lapse seismic difference data and decrease detectability. One of these factors is random noise. In this paper, we propose five different methods aimed at improving the quality and detectability of noisy time-lapse seismic difference data. Common to these methods is the transform of the differences to a domain where the time-lapse signal and random noise are well separated. Our proposed methods include direct Fourier transform based spectral decomposition, bispectra, wavelet transform, singular value decomposition and hybrid methods. We also propose a method that combines multiple time-lapse difference data and gives a final difference which enhances the common part and attenuates the differences of the multiple difference images resulting in a better detectability than the original images. A synthetic time-lapse model is used to demonstrate the feasibility of our proposed methods

  6. Construction of an efficient two-photon fluorescent probe for imaging nitroreductase in live cells and tissues

    Science.gov (United States)

    Zhou, Liyi; Gong, Liang; Hu, Shunqin

    2018-06-01

    Compared with traditional confocal microscopy, two-photon fluorescence microscopy (TPFM), which excites a two-photon (TP) fluorophore by near-infrared light, provides improved three-dimensional image resolution with increased tissue-image depth (>500 μm) and an extended observation time. Therefore, the development of novel functional TP fluorophores has attracted great attention in recent years. Herein, a novel TP fluorophore CM-NH2, which have the donor-π-acceptor (D-π-A)-structure, was designed and synthesized. We further used this dye developed a new type of TP fluorescent probe CM-NO2 for detecting nitroreductase (NTR). Upon incubated with NTR for 15 min, CM-NO2 displayed a 90-fold fluorescence enhancement at 505 nm and the maximal TP action cross-section value after reaction was detected and calculated to be 200 GM at 760 nm. The probe exhibited excellent properties such as high sensitivity, high selectivity, low cytotoxicity, and high photostability. Moreover, the probe was utilized to image the tumor hypoxia in live HeLa cells. Finally, using the CM-NO2 to image NTR in tissues was demonstrated.

  7. Extended two-photon microscopy in live samples with Bessel beams: steadier focus, faster volume scans, and simpler stereoscopic imaging.

    Science.gov (United States)

    Thériault, Gabrielle; Cottet, Martin; Castonguay, Annie; McCarthy, Nathalie; De Koninck, Yves

    2014-01-01

    Two-photon microscopy has revolutionized functional cellular imaging in tissue, but although the highly confined depth of field (DOF) of standard set-ups yields great optical sectioning, it also limits imaging speed in volume samples and ease of use. For this reason, we recently presented a simple and retrofittable modification to the two-photon laser-scanning microscope which extends the DOF through the use of an axicon (conical lens). Here we demonstrate three significant benefits of this technique using biological samples commonly employed in the field of neuroscience. First, we use a sample of neurons grown in culture and move it along the z-axis, showing that a more stable focus is achieved without compromise on transverse resolution. Second, we monitor 3D population dynamics in an acute slice of live mouse cortex, demonstrating that faster volumetric scans can be conducted. Third, we acquire a stereoscopic image of neurons and their dendrites in a fixed sample of mouse cortex, using only two scans instead of the complete stack and calculations required by standard systems. Taken together, these advantages, combined with the ease of integration into pre-existing systems, make the extended depth-of-field imaging based on Bessel beams a strong asset for the field of microscopy and life sciences in general.

  8. Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

    Energy Technology Data Exchange (ETDEWEB)

    Mueller-Harvey, Irene, E-mail: i.mueller-harvey@reading.ac.uk [Chemistry and Biochemistry Laboratory, Food Production and Quality Research Division, School of Agriculture, Policy and Development, University of Reading, P O Box 236, Reading RG6 6AT (United Kingdom); Feucht, Walter, E-mail: walter.feucht@gmail.com [Department of Plant Sciences, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Polster, Juergen, E-mail: j.polster@wzw.tum.de [Department of Physical Biochemistry, Technical University of Munich (TUM), Wissenschaftszentrum Weihenstephan (WZW), D-85354 Freising (Germany); Trnkova, Lucie, E-mail: lucie.trnkova@uhk.cz [University of Hradec Kralove, Faculty of Science, Department of Chemistry, Rokitanskeho 62, 50003 Hradec Kralove (Czech Republic); Burgos, Pierre, E-mail: pierre.burgos@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Parker, Anthony W., E-mail: tony.parker@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom); Botchway, Stanley W., E-mail: stan.botchway@stfc.ac.uk [Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell-Oxford, Didcot, Oxfordshire, OX11 0QX (United Kingdom)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer This fluorescence lifetime imaging microscopy (FLIM) technique for flavanols overcomes autofluorescence interference in cells. Black-Right-Pointing-Pointer Plant flavanols differed in their lifetimes. Black-Right-Pointing-Pointer Dissolved and bound flavanols revealed contrasting lifetime changes. Black-Right-Pointing-Pointer This technique will allow studying of flavanol trafficking in live cells. - Abstract: Two-photon excitation enabled for the first time the observation and measurement of excited state fluorescence lifetimes from three flavanols in solution, which were {approx}1.0 ns for catechin and epicatechin, but <45 ps for epigallocatechin gallate (EGCG). The shorter lifetime for EGCG is in line with a lower fluorescence quantum yield of 0.003 compared to catechin (0.015) and epicatechin (0.018). In vivo experiments with onion cells demonstrated that tryptophan and quercetin, which tend to be major contributors of background fluorescence in plant cells, have sufficiently low cross sections for two-photon excitation at 630 nm and therefore do not interfere with detection of externally added or endogenous flavanols in Allium cepa or Taxus baccata cells. Applying two-photon excitation to flavanols enabled 3-D fluorescence lifetime imaging microscopy and showed that added EGCG penetrated the whole nucleus of onion cells. Interestingly, EGCG and catechin showed different lifetime behaviour when bound to the nucleus: EGCG lifetime increased from <45 to 200 ps, whilst catechin lifetime decreased from 1.0 ns to 500 ps. Semi-quantitative measurements revealed that the relative ratios of EGCG concentrations in nucleoli associated vesicles: nucleus: cytoplasm were ca. 100:10:1. Solution experiments with catechin, epicatechin and histone proteins provided preliminary evidence, via the appearance of a second lifetime ({tau}{sub 2} = 1.9-3.1 ns), that both flavanols may be interacting with histone proteins. We conclude that there

  9. In vivo reactive neural plasticity investigation by means of correlative two photon: electron microscopy

    Science.gov (United States)

    Allegra Mascaro, A. L.; Cesare, P.; Sacconi, L.; Grasselli, G.; Mandolesi, G.; Maco, B.; Knott, G.; Huang, L.; De Paola, V.; Strata, P.; Pavone, F. S.

    2013-02-01

    In the adult nervous system, different populations of neurons correspond to different regenerative behavior. Although previous works showed that olivocerebellar fibers are capable of axonal regeneration in a suitable environment as a response to injury1, we have hitherto no details about the real dynamics of fiber regeneration. We set up a model of singularly axotomized climbing fibers (CF) to investigate their reparative properties in the adult central nervous system (CNS) in vivo. Time lapse two-photon imaging has been combined to laser nanosurgery2, 3 to define a temporal pattern of the degenerative event and to follow the structural rearrangement after injury. To characterize the damage and to elucidate the possible formation of new synaptic contacts on the sprouted branches of the lesioned CF, we combined two-photon in vivo imaging with block face scanning electron microscopy (FIB-SEM). Here we describe the approach followed to characterize the reactive plasticity after injury.

  10. FocusStack and StimServer: a new open source MATLAB toolchain for visual stimulation and analysis of two-photon calcium neuronal imaging data.

    Science.gov (United States)

    Muir, Dylan R; Kampa, Björn M

    2014-01-01

    Two-photon calcium imaging of neuronal responses is an increasingly accessible technology for probing population responses in cortex at single cell resolution, and with reasonable and improving temporal resolution. However, analysis of two-photon data is usually performed using ad-hoc solutions. To date, no publicly available software exists for straightforward analysis of stimulus-triggered two-photon imaging experiments. In addition, the increasing data rates of two-photon acquisition systems imply increasing cost of computing hardware required for in-memory analysis. Here we present a Matlab toolbox, FocusStack, for simple and efficient analysis of two-photon calcium imaging stacks on consumer-level hardware, with minimal memory footprint. We also present a Matlab toolbox, StimServer, for generation and sequencing of visual stimuli, designed to be triggered over a network link from a two-photon acquisition system. FocusStack is compatible out of the box with several existing two-photon acquisition systems, and is simple to adapt to arbitrary binary file formats. Analysis tools such as stack alignment for movement correction, automated cell detection and peri-stimulus time histograms are already provided, and further tools can be easily incorporated. Both packages are available as publicly-accessible source-code repositories.

  11. Two-photon excited fluorescence spectroscopy and imaging of melanin in vitro and in vivo

    Science.gov (United States)

    Krasieva, Tatiana B.; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Tromberg, Bruce J.

    2012-03-01

    The ability to detect early melanoma non-invasively would improve clinical outcome and reduce mortality. Recent advances in two-photon excited fluorescence (TPEF) in vivo microscopy offer a powerful tool in early malignant melanoma diagnostics. The goal of this work was to develop a TPEF optical index for measuring relative concentrations of eumelanin and pheomelanin since ex vivo studies show that changes in this ratio have been associated with malignant transformation. We acquired TPEF emission spectra (λex=1000 nm) of melanin from several specimens, including human hair, malignant melanoma cell lines, and normal melanocytes and keratinocytes in different skin layers (epidermis, papillary dermis) in five healthy volunteers in vivo. We found that the pheomelanin emission peaks at around 620 nm and is blue-shifted from the eumelanin with broad maximum at 640-680nm. We defined "optical melanin index" (OMI) as a ratio of fluorescence signal intensities measured at 645 nm and 615nm. The measured OMI for a melanoma cell line MNT-1 was 1.6+/-0.2. The MNT-46 and MNT-62 lines (Mc1R gene knockdown) showed an anticipated change in melanins production ratio and had OMI of 0.55+/-0.05 and 0.17+/-0.02, respectively, which strongly correlated with HPLC data obtained for these lines. Average OMI measured for basal cells layers (melanocytes and keratinocytes) in normal human skin type I, II-III (not tanned and tanned) in vivo was 0.5, 1.05 and 1.16 respectively. We could not dependably detect the presence of pheomelanin in highly pigmented skin type V-VI. These data suggest that a non-invasive TPEF index could potentially be used for rapid melanin ratio characterization both in vitro and in vivo, including pigmented lesions.

  12. Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment

    Directory of Open Access Journals (Sweden)

    Clément eRicard

    2014-02-01

    Full Text Available The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate of each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology.

  13. Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment.

    Science.gov (United States)

    Ricard, Clément; Debarbieux, Franck Christian

    2014-01-01

    The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM) model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color) images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology.

  14. Two-photon microscopy for imaging of the (atherosclerotic) vascular wall: a proof of concept study

    NARCIS (Netherlands)

    van Zandvoort, Marc; Engels, Wim; Douma, Kim; Beckers, Linda; Oude Egbrink, Mirjam; Daemen, Mat; Slaaf, Dick W.

    2004-01-01

    Understanding atherogenesis will benefit significantly from simultaneous imaging, both ex vivo and in vivo, of structural and functional information at the (sub)cellular level within intact arteries. Due to limited penetration depth and loss of resolution with depth, intravital and confocal

  15. Scanless functional imaging of hippocampal networks using patterned two-photon illumination through GRIN lenses

    KAUST Repository

    Moretti, Claudio; Antonini, Andrea; Bovetti, Serena; Liberale, Carlo; Fellin, Tommaso

    2016-01-01

    functional imaging in rodent hippocampal networks in vivo ~1.2 mm below the brain surface. Our results open the way to the application of patterned illumination approaches to deep regions of highly scattering biological tissues, such as the mammalian brain.

  16. Integrated femtosecond stimulated Raman scattering and two-photon fluorescence imaging of subcellular lipid and vesicular structures

    Science.gov (United States)

    Li, Xuesong; Lam, Wen Jiun; Cao, Zhe; Hao, Yan; Sun, Qiqi; He, Sicong; Mak, Ho Yi; Qu, Jianan Y.

    2015-11-01

    The primary goal of this study is to demonstrate that stimulated Raman scattering (SRS) as a new imaging modality can be integrated into a femtosecond (fs) nonlinear optical (NLO) microscope system. The fs sources of high pulse peak power are routinely used in multimodal nonlinear microscopy to enable efficient excitation of multiple NLO signals. However, with fs excitations, the SRS imaging of subcellular lipid and vesicular structures encounters significant interference from proteins due to poor spectral resolution and a lack of chemical specificity, respectively. We developed a unique NLO microscope of fs excitation that enables rapid acquisition of SRS and multiple two-photon excited fluorescence (TPEF) signals. In the in vivo imaging of transgenic C. elegans animals, we discovered that by cross-filtering false positive lipid signals based on the TPEF signals from tryptophan-bearing endogenous proteins and lysosome-related organelles, the imaging system produced highly accurate assignment of SRS signals to lipid. Furthermore, we demonstrated that the multimodal NLO microscope system could sequentially image lipid structure/content and organelles, such as mitochondria, lysosomes, and the endoplasmic reticulum, which are intricately linked to lipid metabolism.

  17. Inter- and intra-observer variability of time-lapse annotations

    DEFF Research Database (Denmark)

    Sundvall, Linda; Ingerslev, Hans Jakob; Breth Knudsen, Ulla

    2013-01-01

    . This provides the basis for further investigation of embryo assessment and selection by time-lapse imaging in prospective trials. Study funding/competing interest(s): Research at the Fertility Clinic was funded by an unrestricted grant from Ferring and MSD. The authors have no competing interests to declare.......Study question: How consistent is the time-lapse annotation of dynamic and static morphologic parameters of embryo development, within and between observers? Summary answer: The assessment of dynamic parameters is characterized by almost perfect agreement within and between observers. What is known...... already: The commonly employed method used to assess embryos in IVF treatments is based on static evaluation of morphology in a microscope, but this is limited by substantial intra- and inter-observer variation. Time-lapse imaging has been proposed as a method to refine embryo selection by adding new...

  18. Time-Lapse Videos for Physics Education: Specific Examples

    Science.gov (United States)

    Vollmer, Michael; Möllmann, Klaus-Peter

    2018-01-01

    There are many physics experiments with long time scales such that they are usually neither shown in the physics class room nor in student labs. However, they can be easily recorded with time-lapse cameras and the respective time-lapse videos allow qualitative and/or quantitative analysis of the underlying physics. Here, we present some examples…

  19. Time-lapse video sysem used to study nesting gyrfalcons

    Science.gov (United States)

    Booms, Travis; Fuller, Mark R.

    2003-01-01

    We used solar-powered time-lapse video photography to document nesting Gyrfalcon (Falco rusticolus) food habits in central West Greenland from May to July in 2000 and 2001. We collected 2677.25 h of videotape from three nests, representing 94, 87, and 49% of the nestling period at each nest. The video recorded 921 deliveries of 832 prey items. We placed 95% of the items into prey categories. The image quality was good but did not reveal enough detail to identify most passerines to species. We found no evidence that Gyrfalcons were negatively affected by the video system after the initial camera set-up. The video system experienced some mechanical problems but proved reliable. The system likely can be used to effectively document the food habits and nesting behavior of other birds, especially those delivering large prey to a nest or other frequently used site.

  20. Dynamic characterization of hydrophobic and hydrophilic solutes in oleic-acid enhanced transdermal delivery using two-photon fluorescence microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tseng, Te-Yu; Yang, Chiu-Sheng; Chen, Yang-Fang [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Tsai, Tsung-Hua [Department of Dermatology, Far Eastern Memorial Hospital, New Taipei City, Taiwan (China); Dong, Chen-Yuan, E-mail: cydong@phys.ntu.edu.tw [Department of Physics, National Taiwan University, Taipei, Taiwan (China); Center for Quantum Science and Engineering, National Taiwan University, Taipei, Taiwan (China); Center for Optoelectronic Biomedicine, National Taiwan University, Taipei, Taiwan (China)

    2014-10-20

    In this letter, we propose an efficient methodology of investigating dynamic properties of sulforhodamine B and rhodamine B hexyl ester molecules transporting across ex-vivo human stratum corneum with and without oleic acid enhancement. Three-dimensional, time-lapse fluorescence images of the stratum corneum can be obtained using two-photon fluorescence microscopy. Furthermore, temporal quantifications of transport enhancements in diffusion parameters can be achieved with the use of Fick's second law. Dynamic characterization of solutes transporting across the stratum corneum is an effective method for understanding transient phenomena in transdermal delivery of probe molecules, leading to improved delivery strategies of molecular species for therapeutic purposes.

  1. Volumetric label-free imaging and 3D reconstruction of mammalian cochlea based on two-photon excitation fluorescence microscopy

    International Nuclear Information System (INIS)

    Zhang, Xianzeng; Zhan, Zhenlin; Xie, Shusen; Geng, Yang; Ye, Qing

    2013-01-01

    The visualization of the delicate structure and spatial relationship of intracochlear sensory cells has relied on the laborious procedures of tissue excision, fixation, sectioning and staining for light and electron microscopy. Confocal microscopy is advantageous for its high resolution and deep penetration depth, yet disadvantageous due to the necessity of exogenous labeling. In this study, we present the volumetric imaging of rat cochlea without exogenous dyes using a near-infrared femtosecond laser as the excitation mechanism and endogenous two-photon excitation fluorescence (TPEF) as the contrast mechanism. We find that TPEF exhibits strong contrast, allowing cellular and even subcellular resolution imaging of the cochlea, differentiating cell types, visualizing delicate structures and the radial nerve fiber. Our results further demonstrate that 3D reconstruction rendered with z-stacks of optical sections enables better revealment of fine structures and spatial relationships, and easily performed morphometric analysis. The TPEF-based optical biopsy technique provides great potential for new and sensitive diagnostic tools for hearing loss or hearing disorders, especially when combined with fiber-based microendoscopy. (paper)

  2. NEAR-IR TWO PHOTON MICROSCOPY IMAGING OF SILICA NANOPARTICLES FUNCTIONALIZED WITH ISOLATED SENSITIZED Yb(III) CENTERS

    Energy Technology Data Exchange (ETDEWEB)

    Lapadula, Giuseppe; Bourdolle, Adrien; Allouche, Florian; Conley, Matthew P.; Maron, Laurent; Lukens, Wayne W.; Guyot, Yannick; Andraud, Chantal; Brasselet, Sophie; Copé; ret, Christophe; Maury, Olivier; Andersen, Richard A.

    2013-01-12

    Bright nano objects emitting in the near infrared with a maximal cross section of 41.4 x 103 GM (Goppert Mayer), were prepared by implanting ca. 180 4,4 diethylaminostyryl 2,2 bipyridine (DEAS) Yb(III) complexes on the surface of 12 nm silica nanoparticles. The surface complexes ([DEAS Ln SiO2], Ln =Y,Yb) were characterized using IR, solid state NMR, UV Vis, EXAFS spectroscopies in combination with the preparation and characterization of similar molecular analogues by analytical techniques (IR, solution NMR, UV Vis, X ray crystallography) as well as DFT calculations. Starting from the partial dehydroxylation of the silica at 700 C on high vacuum having 0.8 OH.nm 2, the grafting of Ln(N(SiMe3)2)3 generate ≤SiO Ln(N(SiMe3)2)2, which upon thermal step and coordination of the DEAS chromophore yields (≤SiO)3Ln(DEAS). Surface and molecular analogues display similar properties, in terms of DEAS binding constants absorption maxima and luminescence properties (intense emission band assigned to a ligand centered CT fluorescence and life time) in the solid state, consistent with the molecular nature of the surface species. The densely functionalized nanoparticles can be dispersed via ultra-sonication in small ca. 15-20 nm aggregates (1 to 6 elementary particles) that were detected using two photon microscopy imaging at 720 nm excitation, making them promising nano objects for bio imaging.

  3. Femtosecond, two-photon laser-induced-fluorescence imaging of atomic oxygen in an atmospheric-pressure plasma jet

    Science.gov (United States)

    Schmidt, Jacob B.; Sands, Brian L.; Kulatilaka, Waruna D.; Roy, Sukesh; Scofield, James; Gord, James R.

    2015-06-01

    Femtosecond, two-photon-absorption laser-induced-fluorescence (fs-TALIF) spectroscopy is employed to measure space- and time-resolved atomic-oxygen distributions in a nanosecond, repetitively pulsed, externally grounded, atmospheric-pressure plasma jet flowing helium with a variable oxygen admixture. The high-peak-intensity, low-average-energy femtosecond pulses result in increased TALIF signal with reduced photolytic inferences. This allows 2D imaging of absolute atomic-oxygen number densities ranging from 5.8   ×   1015 to 2.0   ×   1012cm-3 using a cooled CCD with an external intensifier. Xenon is used for signal and imaging-system calibrations to quantify the atomic-oxygen fluorescence signal. Initial results highlight a transition in discharge morphology from annular to filamentary, corresponding with a change in plasma chemistry from ozone to atomic oxygen production, as the concentration of oxygen in the feed gas is changed at a fixed voltage-pulse-repetition rate. In this configuration, significant concentrations of reactive oxygen species may be remotely generated by sustaining an active discharge beyond the confines of the dielectric capillary, which may benefit applications that require large concentrations of reactive oxygen species such as material processing or biomedical devices.

  4. Mercury effects on Thalassiosira weissflogii: Applications of two-photon excitation chlorophyll fluorescence lifetime imaging and flow cytometry

    International Nuclear Information System (INIS)

    Wu Yun; Zeng Yan; Qu, Jianan Y.; Wang Wenxiong

    2012-01-01

    The toxic effects of inorganic mercury [Hg(II)] and methylmercury (MeHg) on the photosynthesis and population growth in a marine diatom Thalassiosira weissflogii were investigated using two methods: two-photon excitation fluorescence lifetime imaging (FLIM) and flow cytometry (FCM). For photosynthesis, Hg(II) exposure increased the average chlorophyll fluorescence lifetime, whereas such increment was not found under MeHg stress. This may be caused by the inhibitory effect of Hg(II) instead of MeHg on the electron transport chain. For population growth, modeled specific growth rate data showed that the reduction in population growth by Hg(II) mainly resulted from an increased number of injured cells, while the live cells divided at the normal rates. However, MeHg inhibitory effects on population growth were contributed by the reduced division rates of all cells. Furthermore, the cell images and the FCM data reflected the morphological changes of diatom cells under Hg(II)/MeHg exposure vividly and quantitatively. Our results demonstrated that the toxigenicity mechanisms between Hg(II) and MeHg were different in the algal cells.

  5. Calcium rubies: a family of red-emitting functionalizable indicators suitable for two-photon Ca2+ imaging.

    Science.gov (United States)

    Collot, Mayeul; Loukou, Christina; Yakovlev, Aleksey V; Wilms, Christian D; Li, Dongdong; Evrard, Alexis; Zamaleeva, Alsu; Bourdieu, Laurent; Léger, Jean-François; Ropert, Nicole; Eilers, Jens; Oheim, Martin; Feltz, Anne; Mallet, Jean-Maurice

    2012-09-12

    We designed Calcium Rubies, a family of functionalizable BAPTA-based red-fluorescent calcium (Ca(2+)) indicators as new tools for biological Ca(2+) imaging. The specificity of this Ca(2+)-indicator family is its side arm, attached on the ethylene glycol bridge that allows coupling the indicator to various groups while leaving open the possibility of aromatic substitutions on the BAPTA core for tuning the Ca(2+)-binding affinity. Using this possibility we now synthesize and characterize three different CaRubies with affinities between 3 and 22 μM. Their long excitation and emission wavelengths (peaks at 586/604 nm) allow their use in otherwise challenging multicolor experiments, e.g., when combining Ca(2+) uncaging or optogenetic stimulation with Ca(2+) imaging in cells expressing fluorescent proteins. We illustrate this capacity by the detection of Ca(2+) transients evoked by blue light in cultured astrocytes expressing CatCh, a light-sensitive Ca(2+)-translocating channelrhodopsin linked to yellow fluorescent protein. Using time-correlated single-photon counting, we measured fluorescence lifetimes for all CaRubies and demonstrate a 10-fold increase in the average lifetime upon Ca(2+) chelation. Since only the fluorescence quantum yield but not the absorbance of the CaRubies is Ca(2+)-dependent, calibrated two-photon fluorescence excitation measurements of absolute Ca(2+) concentrations are feasible.

  6. Two-photon physics

    International Nuclear Information System (INIS)

    Bardeen, W.A.

    1981-10-01

    A new experimental frontier has recently been opened to the study of two photon processes. The first results of many aspects of these reactions are being presented at this conference. In contrast, the theoretical development of research ito two photon processes has a much longer history. This talk reviews the many different theoretical ideas which provide a detailed framework for our understanding of two photon processes

  7. Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging.

    Science.gov (United States)

    Chen, Bin; Feng, Guangxue; He, Bairong; Goh, Chiching; Xu, Shidang; Ramos-Ortiz, Gabriel; Aparicio-Ixta, Laura; Zhou, Jian; Ng, Laiguan; Zhao, Zujin; Liu, Bin; Tang, Ben Zhong

    2016-02-10

    Robust luminescent dyes with efficient two-photon fluorescence are highly desirable for biological imaging applications, but those suitable for organic dots fabrication are still rare because of aggregation-caused quenching. In this work, a red fluorescent silole, 2,5-bis[5-(dimesitylboranyl)thiophen-2-yl]-1-methyl-1,3,4-triphenylsilole ((MesB)2 DTTPS), is synthesized and characterized. (MesB)2 DTTPS exhibits enhanced fluorescence efficiency in nanoaggregates, indicative of aggregation-enhanced emission (AEE). The organic dots fabricated by encapsulating (MesB)2 DTTPS within lipid-PEG show red fluorescence peaking at 598 nm and a high fluorescence quantum yield of 32%. Upon excitation at 820 nm, the dots show a large two-photon absorption cross section of 3.43 × 10(5) GM, which yields a two-photon action cross section of 1.09 × 10(5) GM. These (MesB)2 DTTPS dots show good biocompatibility and are successfully applied to one-photon and two-photon fluorescence imaging of MCF-7 cells and two-photon in vivo visualization of the blood vascular of mouse muscle in a high-contrast and noninvasive manner. Moreover, the 3D blood vasculature located at the mouse ear skin with a depth of over 100 μm can also be visualized clearly, providing the spatiotemporal information about the whole blood vascular network. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Inter and intra-observer variability of time-lapse annotations

    DEFF Research Database (Denmark)

    Sundvall Germeys, Linda Karin M; Ingerslev, Hans Jakob; Knudsen, Ulla Breth

    . This provides the basis for further investigation of embryo assessment and selection by time-lapse imaging in prospective trials. Study funding/competing interest(s): Research at the Fertility Clinic was funded by an unrestricted grant from Ferring and MSD. The authors have no competing interests to declare....

  9. Latest time-lapse seismic data from Sleipner yield new insights into CO2 plume development

    NARCIS (Netherlands)

    Chadwick, R.A.; Noy, D.; Arts, R.; Eiken, O.

    2009-01-01

    Since its inception in 1996, the CO2 injection operation at Sleipner has been monitored by 3D time-lapse seismic surveys. Striking images of the CO2 plume have been obtained, showing a multi-tier feature of high reflectivity, interpreted as arising from a number of thin layers of CO2 trapped beneath

  10. Time-lapse videos for physics education: specific examples

    Science.gov (United States)

    Vollmer, Michael; Möllmann, Klaus-Peter

    2018-05-01

    There are many physics experiments with long time scales such that they are usually neither shown in the physics class room nor in student labs. However, they can be easily recorded with time-lapse cameras and the respective time-lapse videos allow qualitative and/or quantitative analysis of the underlying physics. Here, we present some examples from thermal physics (melting, evaporation, cooling) as well as diffusion processes

  11. Two-photon imaging of formaldehyde in live cells and animals utilizing a lysosome-targetable and acidic pH-activatable fluorescent probe.

    Science.gov (United States)

    Xie, Xilei; Tang, Fuyan; Shangguan, Xiaoyan; Che, Shiyi; Niu, Jinye; Xiao, Yongsheng; Wang, Xu; Tang, Bo

    2017-06-13

    Lyso-TPFP presents lysosomal targetability and an acidic pH-activatable response toward formaldehyde. Thus, it exclusively visualizes lysosomal formaldehyde and is immune against it in neutral cytosol and other organelles. In addition, two-photon fluorescence imaging endows Lyso-TPFP with the capability of in situ tracking formaldehyde in live cells and animals.

  12. Direct Vpr-Vpr Interaction in Cells monitored by two Photon Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging

    Directory of Open Access Journals (Sweden)

    Mély Yves

    2008-09-01

    Full Text Available Abstract Background The human immunodeficiency virus type 1 (HIV-1 encodes several regulatory proteins, notably Vpr which influences the survival of the infected cells by causing a G2/M arrest and apoptosis. Such an important role of Vpr in HIV-1 disease progression has fuelled a large number of studies, from its 3D structure to the characterization of specific cellular partners. However, no direct imaging and quantification of Vpr-Vpr interaction in living cells has yet been reported. To address this issue, eGFP- and mCherry proteins were tagged by Vpr, expressed in HeLa cells and their interaction was studied by two photon fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy. Results Results show that Vpr forms homo-oligomers at or close to the nuclear envelope. Moreover, Vpr dimers and trimers were found in the cytoplasm and in the nucleus. Point mutations in the three α helices of Vpr drastically impaired Vpr oligomerization and localization at the nuclear envelope while point mutations outside the helical regions had no effect. Theoretical structures of Vpr mutants reveal that mutations within the α-helices could perturb the leucine zipper like motifs. The ΔQ44 mutation has the most drastic effect since it likely disrupts the second helix. Finally, all Vpr point mutants caused cell apoptosis suggesting that Vpr-mediated apoptosis functions independently from Vpr oligomerization. Conclusion We report that Vpr oligomerization in HeLa cells relies on the hydrophobic core formed by the three α helices. This oligomerization is required for Vpr localization at the nuclear envelope but not for Vpr-mediated apoptosis.

  13. Probabilistic 3-D time-lapse inversion of magnetotelluric data: application to an enhanced geothermal system

    Science.gov (United States)

    Rosas-Carbajal, M.; Linde, N.; Peacock, J.; Zyserman, F. I.; Kalscheuer, T.; Thiel, S.

    2015-12-01

    Surface-based monitoring of mass transfer caused by injections and extractions in deep boreholes is crucial to maximize oil, gas and geothermal production. Inductive electromagnetic methods, such as magnetotellurics, are appealing for these applications due to their large penetration depths and sensitivity to changes in fluid conductivity and fracture connectivity. In this work, we propose a 3-D Markov chain Monte Carlo inversion of time-lapse magnetotelluric data to image mass transfer following a saline fluid injection. The inversion estimates the posterior probability density function of the resulting plume, and thereby quantifies model uncertainty. To decrease computation times, we base the parametrization on a reduced Legendre moment decomposition of the plume. A synthetic test shows that our methodology is effective when the electrical resistivity structure prior to the injection is well known. The centre of mass and spread of the plume are well retrieved. We then apply our inversion strategy to an injection experiment in an enhanced geothermal system at Paralana, South Australia, and compare it to a 3-D deterministic time-lapse inversion. The latter retrieves resistivity changes that are more shallow than the actual injection interval, whereas the probabilistic inversion retrieves plumes that are located at the correct depths and oriented in a preferential north-south direction. To explain the time-lapse data, the inversion requires unrealistically large resistivity changes with respect to the base model. We suggest that this is partly explained by unaccounted subsurface heterogeneities in the base model from which time-lapse changes are inferred.

  14. N-doped carbon dots derived from bovine serum albumin and formic acid with one- and two-photon fluorescence for live cell nuclear imaging.

    Science.gov (United States)

    Tan, Mingqian; Li, Xintong; Wu, Hao; Wang, Beibei; Wu, Jing

    2015-12-01

    Carbon dots with both one- and two-photon fluorescence have drawn great attention for biomedical imaging. Herein, nitrogen-doped carbon dots were facilely developed by one-pot hydrothermal method using bovine serum albumin and formic acid as carbon sources. They are highly water-soluble with strong fluorescence when excited with ultraviolet or near infrared light. The carbon dots have a diameter of ~8.32 nm and can emit strong two-photon induced fluorescence upon excitation at 750 nm with a femtosecond laser. X-ray photoelectron spectrometer analysis revealed that the carbon dots contained three components, C, N and O, corresponding to the peak at 285, 398 and 532 eV, respectively. The Fourier-transform infrared spectroscopy analysis revealed that there are carboxyl and carboxylic groups on the surface, which allowed further linking of functional molecules. pH stability study demonstrated that the carbon dots are able to be used in a wide range of pH values. The fluorescence mechanism is also discussed in this study. Importantly, these carbon dots are biocompatible and highly photostable, which can be directly applied for both one- and two-photon living cell imaging. After proper surface functionalization with TAT peptide, they can be used as fluorescent probes for live cell nuclear-targeted imaging. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Tale of two photons

    International Nuclear Information System (INIS)

    Anon.

    1994-01-01

    A very profitable spinoff from electron- positron collisions is two-photon physics. Rather than the electron and positron interacting directly via an exchanged photon, two virtual (transient) photons, one from each particle, get tangled up. With new electron-positron colliders appearing on the scene, a topical meeting on two-photon physics - 'From DAPHNE to LEP 200 and beyond' - held from 2-4 February in Paris, in the premises of the Ministry of Higher Education and Research, was particularly timely. Some 60 physicists, both experimentalists and theorists, participated, with some thirty speakers

  16. Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

    Science.gov (United States)

    Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

    2014-06-01

    Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

  17. Time-lapse controlled-source electromagnetics using interferometry

    NARCIS (Netherlands)

    Hunziker, J.W.; Slob, E.C.; Wapenaar, C.P.A.

    In time-lapse controlled-source electromagnetics, it is crucial that the source and the receivers are positioned at exactly the same location at all times of measurement. We use interferometry by multidimensional deconvolution (MDD) to overcome problems in repeatability of the source location.

  18. Multifunctional biocompatible graphene oxide quantum dots decorated magnetic nanoplatform for efficient capture and two-photon imaging of rare tumor cells.

    Science.gov (United States)

    Shi, Yongliang; Pramanik, Avijit; Tchounwou, Christine; Pedraza, Francisco; Crouch, Rebecca A; Chavva, Suhash Reddy; Vangara, Aruna; Sinha, Sudarson Sekhar; Jones, Stacy; Sardar, Dhiraj; Hawker, Craig; Ray, Paresh Chandra

    2015-05-27

    Circulating tumor cells (CTCs) are extremely rare cells in blood containing billions of other cells. The selective capture and identification of rare cells with sufficient sensitivity is a real challenge. Driven by this need, this manuscript reports the development of a multifunctional biocompatible graphene oxide quantum dots (GOQDs) coated, high-luminescence magnetic nanoplatform for the selective separation and diagnosis of Glypican-3 (GPC3)-expressed Hep G2 liver cancer tumor CTCs from infected blood. Experimental data show that an anti-GPC3-antibody-attached multifunctional nanoplatform can be used for selective Hep G2 hepatocellular carcinoma tumor cell separation from infected blood containing 10 tumor cells/mL of blood in a 15 mL sample. Reported data indicate that, because of an extremely high two-photon absorption cross section (40530 GM), an anti-GPC3-antibody-attached GOQDs-coated magnetic nanoplatform can be used as a two-photon luminescence platform for selective and very bright imaging of a Hep G2 tumor cell in a biological transparency window using 960 nm light. Experimental results with nontargeted GPC3(-) and SK-BR-3 breast cancer cells show that multifunctional-nanoplatform-based cell separation, followed by two-photon imaging, is highly selective for Hep G2 hepatocellular carcinoma tumor cells.

  19. A fast-response two-photon fluorescent probe for imaging endogenous H2O2 in living cells and tissues

    Science.gov (United States)

    Lu, Yanan; Shi, Xiaomin; Fan, Wenlong; Black, Cory A.; Lu, Zhengliang; Fan, Chunhua

    2018-02-01

    As a second messenger, hydrogen peroxide plays significant roles in numerous physiological and pathological processes and is related to various diseases including inflammatory disease, diabetes, neurodegenerative disorders, cardiovascular disease and Alzheimer's disease. Two-photon (TP) fluorescent probes reported for the detection of endogenous H2O2 are rare and most have drawbacks such as slow response and low sensitivity. In this report, we demonstrate a simple H2O2-specific TP fluorescent probe (TX-HP) containing a two-photon dye 6-hydroxy-2,3,4,4a-tetrahydro-1H-xanthen-1-one (TX) on the modulation of the ICT process. The probe exhibits a rapid fluorescent response to H2O2 in 9 min with both high sensitivity and selectivity. The probe can detect exogenous H2O2 in living cells. Furthermore, the probe is successfully utilized for imaging H2O2 in liver tissues.

  20. Two Photon Distribution Amplitudes

    International Nuclear Information System (INIS)

    El Beiyad, M.; Pire, B.; Szymanowski, L.; Wallon, S.

    2008-01-01

    The factorization of the amplitude of the process γ*γ→γγ in the low energy and high photon virtuality region is demonstrated at the Born order and in the leading logarithmic approximation. The leading order two photon (generalized) distribution amplitudes exhibit a characteristic ln Q 2 behaviour and obey new inhomogeneous evolution equations

  1. Cell and brain tissue imaging of the flavonoid fisetin using label-free two-photon microscopy.

    Science.gov (United States)

    Krasieva, Tatiana B; Ehren, Jennifer; O'Sullivan, Thomas; Tromberg, Bruce J; Maher, Pamela

    2015-10-01

    Over the last few years, we have identified an orally active, novel neuroprotective and cognition-enhancing molecule, the flavonoid fisetin. Fisetin not only has direct antioxidant activity but it can also increase the intracellular levels of glutathione, the major intracellular antioxidant. Fisetin can also activate key neurotrophic factor signaling pathways. In addition, it has anti-inflammatory activity against microglia and astrocytes and inhibits the activity of lipoxygenases, thereby reducing the production of pro-inflammatory eicosanoids and their by-products. However, key questions about its targets and brain penetration remain. In this study, we used label-free two-photon microscopy of intrinsic fisetin fluorescence to examine the localization of fisetin in living nerve cells and the brains of living mice. In cells, fisetin but not structurally related flavonols with different numbers of hydroxyl groups, localized to the nucleoli suggesting that key targets of fisetin may reside in this organelle. In the mouse brain, following intraperitoneal injection and oral administration, fisetin rapidly distributed to the blood vessels of the brain followed by a slower dispersion into the brain parenchyma. Thus, these results provide further support for the effects of fisetin on brain function. In addition, they suggest that label-free two-photon microscopy may prove useful for studying the intracellular and tissue distribution of other intrinsically-fluorescent flavonoids. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Quantitative evaluation of redox ratio and collagen characteristics during breast cancer chemotherapy using two-photon intrinsic imaging.

    Science.gov (United States)

    Wu, Shulian; Huang, Yudian; Tang, Qinggong; Li, Zhifang; Horng, Hannah; Li, Jiatian; Wu, Zaihua; Chen, Yu; Li, Hui

    2018-03-01

    Preoperative neoadjuvant treatment in locally advanced breast cancer is recognized as an effective adjuvant therapy, as it improves treatment outcomes. However, the potential complications remain a threat, so there is an urgent clinical need to assess both the tumor response and changes in its microenvironment using non-invasive and precise identification techniques. Here, two-photon microscopy was employed to detect morphological alterations in breast cancer progression and recession throughout chemotherapy. The changes in structure were analyzed based on the autofluorescence and collagen of differing statuses. Parameters, including optical redox ratio, the ratio of second harmonic generation and auto-fluorescence signal, collagen density, and collagen shape orientation, were studied. Results indicate that these parameters are potential indicators for evaluating breast tumors and their microenvironment changes during progression and chemotherapy. Combined analyses of these parameters could provide a quantitative, novel method for monitoring tumor therapy.

  3. Growth Rate and Morphology of a Single Calcium Carbonate Crystal on Polysulfone Film Measured with Time Lapse Raman Micro Spectroscopy

    NARCIS (Netherlands)

    Liszka, B.; Lenferink, Aufrid T.M.; Otto, Cornelis

    2016-01-01

    The growth of single, self- nucleated calcium carbonate crystals on a polysulfone (PSU) film was investigated with high resolution, time lapse Raman imaging. The Raman images were acquired on the interface of the polymer with the crystal. The growth of crystals could thus be followed in time. PSU is

  4. Probabilistic 3-D time-lapse inversion of magnetotelluric data: Application to an enhanced geothermal system

    Science.gov (United States)

    Rosas-Carbajal, Marina; Linde, Nicolas; Peacock, Jared R.; Zyserman, F. I.; Kalscheuer, Thomas; Thiel, Stephan

    2015-01-01

    Surface-based monitoring of mass transfer caused by injections and extractions in deep boreholes is crucial to maximize oil, gas and geothermal production. Inductive electromagnetic methods, such as magnetotellurics, are appealing for these applications due to their large penetration depths and sensitivity to changes in fluid conductivity and fracture connectivity. In this work, we propose a 3-D Markov chain Monte Carlo inversion of time-lapse magnetotelluric data to image mass transfer following a saline fluid injection. The inversion estimates the posterior probability density function of the resulting plume, and thereby quantifies model uncertainty. To decrease computation times, we base the parametrization on a reduced Legendre moment decomposition of the plume. A synthetic test shows that our methodology is effective when the electrical resistivity structure prior to the injection is well known. The centre of mass and spread of the plume are well retrieved.We then apply our inversion strategy to an injection experiment in an enhanced geothermal system at Paralana, South Australia, and compare it to a 3-D deterministic time-lapse inversion. The latter retrieves resistivity changes that are more shallow than the actual injection interval, whereas the probabilistic inversion retrieves plumes that are located at the correct depths and oriented in a preferential north-south direction. To explain the time-lapse data, the inversion requires unrealistically large resistivity changes with respect to the base model. We suggest that this is partly explained by unaccounted subsurface heterogeneities in the base model from which time-lapse changes are inferred.

  5. Calibrating vadose zone models with time-lapse gravity data

    DEFF Research Database (Denmark)

    Christiansen, Lars; Binning, Philip John; Rosbjerg, Dan

    2011-01-01

    The vadose zone plays an important role in the hydrologic cycle. Various geophysical methods can determine soil water content variations in time and space in volumes ranging from a few cubic centimeters to several cubic meters. In contrast to the established methods, time-lapse gravity measurements...... of changes in soil water content do not rely on a petrophysical relationship between the measured quantity and the water content but give a direct measure of the mass change in the soil. Only recently has the vadose zone been systematically incorporated when ground-based gravity data are used to infer...... hydrologic information. In this study, changes in the soil water content gave rise to a measurable signal in a forced infiltration experiment on a 107-m2 grassland area. Time-lapse gravity data were able to constrain the van Genuchten soil hydraulic parameters in both a synthetic example and a field...

  6. The Use of Intravital Two-Photon and Thick Section Confocal Imaging to Analyze B Lymphocyte Trafficking in Lymph Nodes and Spleen.

    Science.gov (United States)

    Park, Chung; Hwang, Il-Young; Kehrl, John H

    2018-01-01

    Intravital two-photon laser scanning microscopy (TP-LSM) has allowed the direct observation of immune cells in intact organs of living animals. In the B cell biology field TP-LSM has detailed the movement of B cells in high endothelial venules and during their transmigration into lymph organs; described the movement and positioning of B cells within lymphoid organs; outlined the mechanisms by which antigen is delivered to B cells; observed B cell interacting with T cells, other cell types, and even with pathogens; and delineated the egress of B cells from the lymph node (LN) parenchyma into the efferent lymphatics. As the quality of TP-LSM improves and as new fluorescent probes become available additional insights into B cell behavior and function await new investigations. Yet intravital TP-LSM has some disadvantages including a lower resolution than standard confocal microscopy, a narrow imaging window, and a shallow depth of imaging. We have found that supplementing intravital TP-LSM with conventional confocal microscopy using thick LN sections helps to overcome some of these shortcomings. Here, we describe procedures for visualizing the behavior and trafficking of fluorescently labeled, adoptively transferred antigen-activated B cells within the inguinal LN of live mice using two-photon microscopy. Also, we introduce procedures for fixed thick section imaging using standard confocal microscopy, which allows imaging of fluorescently labeled cells deep in the LN cortex and in the spleen with high resolution.

  7. Definition of "banner clouds" based on time lapse movies

    OpenAIRE

    Schween , J. H.; Kuettner , J.; Reinert , D.; Reuder , J.; Wirth , V.

    2007-01-01

    International audience; Banner clouds appear on the leeward side of a mountain and resemble a banner or a flag. This article provides a comprehensive definition of "banner clouds". It is based primarily on an extensive collection of time lapse movies, but previous attempts at an explanation of this phenomenon are also taken into account. The following ingredients are considered essential: the cloud must be attached to the mountain but not appear on the windward side; the cloud must originate ...

  8. Water soluble two-photon fluorescent organic probes for long-term imaging of lysosomes in live cells and tumor spheroids.

    Science.gov (United States)

    Kumari, Pratibha; Verma, Sanjay K; Mobin, Shaikh M

    2018-01-11

    The morphological alteration of lysosomes is a powerful indicator of various pathological disorders. In this regard, we have designed and synthesized a new water soluble fluorescent Schiff-base ligand (L-lyso) containing two hydroxyl groups. L-lyso exhibits excellent two-photon properties with tracking of lysosomes in live cells as well as in 3D tumor spheroids. Furthermore, it can label lysosomes for more than 3 days. Thus, L-lyso has an edge over the commercially available expensive LysoTracker probes and also over other reported probes in terms of its long-term imaging, water solubility and facile synthesis.

  9. Two-photon time-lapse microscopy of BODIPY-cholesterol reveals anomalous sterol diffusion in chinese hamster ovary cells

    DEFF Research Database (Denmark)

    Lund, F. W.; Lomholt, M. A.; Solanko, L. M.

    2012-01-01

    to sterol trafficking from the plasma membrane. Significant photobleaching, however, limits the possibilities for in-depth analysis of sterol dynamics using DHE. Co-trafficking studies with DHE and the recently introduced fluorescent cholesterol analog BODIPY-cholesterol (BChol) suggested that the latter...

  10. Time-Lapse Joint Inversion of Cross-Well DC Resistivity and Seismic Data: A Numerical Investigation

    Science.gov (United States)

    Time-lapse joint inversion of geophysical data is required to image the evolution of oil reservoirs during production and enhanced oil recovery, CO2 sequestration, geothermal fields during production, and to monitor the evolution of contaminant plumes. Joint inversion schemes red...

  11. Two-photon imaging during prolonged middle cerebral artery occlusion in mice reveals recovery of dendritic structure after reperfusion.

    Science.gov (United States)

    Li, Ping; Murphy, Timothy H

    2008-11-12

    Filament occlusion of the middle cerebral artery (MCA) is a well accepted animal model of focal ischemia. Advantages of the model are relatively long occlusion times and a large penumbra region that simulates aspects of human stroke. Here, we use two-photon and confocal microscopy in combination with regional measurement of blood flow using laser speckle to assess the spatial relationship between the borders of the MCA ischemic territory and loss of dendrite structure, as well as the effect of reperfusion on dendritic damage in adult YFP (yellow fluorescent protein) and GFP (green fluorescent protein) C57BL/6 transgenic mice with fluorescent (predominantly layer 5) neurons. By examining the spatial extent of dendritic damage, we determined that 60 min of MCA occlusion produced a core with severe structural damage that did not recover after reperfusion (begins approximately 3.8 mm lateral to midline), a reversibly damaged area up to 0.6 mm medial to the core that recovered after reperfusion (penumbra), and a relatively structurally intact area ( approximately 1 mm wide; medial penumbra) with hypoperfusion. Loss of structure was preceded by a single ischemic depolarization 122.1 +/- 10.2 s after occlusion onset. Reperfusion of animals after 60 min of ischemia was not associated with exacerbation of damage (reperfusion injury) and resulted in a significant restoration of blebbed dendritic structure, but only within approximately 0.6 mm lateral of the dendritic damage structural border. In summary, we find that recovery of dendritic structure can occur after reperfusion after even 60 min of ischemia, but is likely restricted to a relatively small penumbra region with partial blood flow or oxygenation.

  12. Label-free imaging of brain and brain tumor specimens with combined two-photon excited fluorescence and second harmonic generation microscopy

    Science.gov (United States)

    Jiang, Liwei; Wang, Xingfu; Wu, Zanyi; Du, Huiping; Wang, Shu; Li, Lianhuang; Fang, Na; Lin, Peihua; Chen, Jianxin; Kang, Dezhi; Zhuo, Shuangmu

    2017-10-01

    Label-free imaging techniques are gaining acceptance within the medical imaging field, including brain imaging, because they have the potential to be applied to intraoperative in situ identifications of pathological conditions. In this paper, we describe the use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) microscopy in combination for the label-free detection of brain and brain tumor specimens; gliomas. Two independently detecting channels were chosen to subsequently collect TPEF/SHG signals from the specimen to increase TPEF/SHG image contrasts. Our results indicate that the combined TPEF/SHG microscopic techniques can provide similar rat brain structural information and produce a similar resolution like conventional H&E staining in neuropathology; including meninges, cerebral cortex, white-matter structure corpus callosum, choroid plexus, hippocampus, striatum, and cerebellar cortex. It can simultaneously detect infiltrating human brain tumor cells, the extracellular matrix collagen fiber of connective stroma within brain vessels and collagen depostion in tumor microenvironments. The nuclear-to-cytoplasmic ratio and collagen content can be extracted as quantitative indicators for differentiating brain gliomas from healthy brain tissues. With the development of two-photon fiberscopes and microendoscope probes and their clinical applications, the combined TPEF and SHG microcopy may become an important multimodal, nonlinear optical imaging approach for real-time intraoperative histological diagnostics of residual brain tumors. These occur in various brain regions during ongoing surgeries through the method of simultaneously identifying tumor cells, and the change of tumor microenvironments, without the need for the removal biopsies and without the need for tissue labelling or fluorescent markers.

  13. Time-lapse seismic waveform modelling and attribute analysis using hydromechanical models for a deep reservoir undergoing depletion

    Science.gov (United States)

    He, Y.-X.; Angus, D. A.; Blanchard, T. D.; Wang, G.-L.; Yuan, S.-Y.; Garcia, A.

    2016-04-01

    Extraction of fluids from subsurface reservoirs induces changes in pore pressure, leading not only to geomechanical changes, but also perturbations in seismic velocities and hence observable seismic attributes. Time-lapse seismic analysis can be used to estimate changes in subsurface hydromechanical properties and thus act as a monitoring tool for geological reservoirs. The ability to observe and quantify changes in fluid, stress and strain using seismic techniques has important implications for monitoring risk not only for petroleum applications but also for geological storage of CO2 and nuclear waste scenarios. In this paper, we integrate hydromechanical simulation results with rock physics models and full-waveform seismic modelling to assess time-lapse seismic attribute resolution for dynamic reservoir characterization and hydromechanical model calibration. The time-lapse seismic simulations use a dynamic elastic reservoir model based on a North Sea deep reservoir undergoing large pressure changes. The time-lapse seismic traveltime shifts and time strains calculated from the modelled and processed synthetic data sets (i.e. pre-stack and post-stack data) are in a reasonable agreement with the true earth models, indicating the feasibility of using 1-D strain rock physics transform and time-lapse seismic processing methodology. Estimated vertical traveltime shifts for the overburden and the majority of the reservoir are within ±1 ms of the true earth model values, indicating that the time-lapse technique is sufficiently accurate for predicting overburden velocity changes and hence geomechanical effects. Characterization of deeper structure below the overburden becomes less accurate, where more advanced time-lapse seismic processing and migration is needed to handle the complex geometry and strong lateral induced velocity changes. Nevertheless, both migrated full-offset pre-stack and near-offset post-stack data image the general features of both the overburden and

  14. Real-time two-photon lithography in controlled flow to create a single-microparticle array and particle-cluster array for optofluidic imaging.

    Science.gov (United States)

    Xu, Bing; Shi, Yang; Lao, Zhaoxin; Ni, Jincheng; Li, Guoqiang; Hu, Yanlei; Li, Jiawen; Chu, Jiaru; Wu, Dong; Sugioka, Koji

    2018-01-30

    Microarray technology provides an excellent platform for biomedical and biochemical research including basic scientific studies, drug discovery, and diagnostics. Here, we develop a novel method referred to as real-time two-photon lithography in a controlled flow in which femtosecond laser two-photon lithography is performed in situ in the sequential mode stopping and flowing the flow of liquid resin containing microparticles to achieve 100% trapping on a one-bead-to-one-trap basis. Polydisperse particles can be all trapped to form a desired array by freely designing trap structures, resulting in an unprecedentedly high capture efficiency of ∼100%. No persistent pressure is needed after trapping which reduces the complexity of the system. In addition, trapping of particle-cluster arrays with a controlled number of particles is also achieved via this method. The trapped particles inside the microchip are successfully applied as microlenses for high quality imaging. The present technology marks an essential step towards a versatile platform for the integration of bead-based assays and paves the way for developing innovative microfluidics, optofluidics, micro-optics and single-cell analysis devices.

  15. Interferometric full-waveform inversion of time-lapse data

    KAUST Repository

    Sinha, Mrinal

    2017-08-17

    One of the key challenges associated with time-lapse surveys is ensuring the repeatability between the baseline and monitor surveys. Non-repeatability between the surveys is caused by varying environmental conditions over the course of different surveys. To overcome this challenge, we propose the use of interferometric full waveform inversion (IFWI) for inverting the velocity model from data recorded by baseline and monitor surveys. A known reflector is used as the reference reflector for IFWI, and the data are naturally redatumed to this reference reflector using natural reflections as the redatuming operator. This natural redatuming mitigates the artifacts introduced by the repeatability errors that originate above the reference reflector.

  16. Quantitative 3-dimensional imaging of auxin and cytokinin levels in transgenic soybean and medicago truncatula roots via two-photon induced fluorescence imaging

    Science.gov (United States)

    Fisher, Jon; Gaillard, Paul; Nurmalasari, Ni Putu Dewi; Fellbaum, Carl; Subramaniam, Sen; Smith, Steve

    2018-02-01

    Industrial nitrogen fertilizers account for nearly 50% of the fossil fuel costs in modern agriculture and contribute to soil and water pollution. Therefore, significant interest exists in understanding and characterizing the efficiency of nitrogen fixation, and the biochemical signaling pathways which orchestrate the plant-microbial symbiosis through which plants fix nitrogen. Legume plant species exhibit a particularly efficient nitrogen uptake mechanism, using root nodules which house nitrogen-fixing rhizobial bacteria. While nodule development has been widely studied, there remain significant gaps in understanding the regulatory hormones' role in plant development. In this work, we produce 3-dimensional maps of auxin (AX) and cytokinin (CK) hormone concentrations within model plant root tips and nodules with respect to root architecture and cell type. Soybean and Medicago plants were transfected with a two-color fluorescent vector with AXsensitive green fluorescent protein (GFP) and CK-sensitive TdTomato (TdT). 3D images of soybean root nodules were captured using two-photon induced fluorescence microscopy. The resulting images were computationally analyzed using the localization code first developed by Weeks and later adapted by Kilfoil, and analyzed in the context of the root architecture. Statistical analysis of the resulting 3D hormone level maps reproduce-well the known roles of AX and CK in developing plant roots, and are the first quantitative description of these regulatory hormones tied to specific plant architecture. The analytical methods used, and the spatial distribution of these key regulatory hormones in plant roots, nodule primordia and root nodules, and their statistical interpretation are presented.

  17. Cardiac muscle organization revealed in 3-D by imaging whole-mount mouse hearts using two-photon fluorescence and confocal microscopy.

    Science.gov (United States)

    Sivaguru, Mayandi; Fried, Glenn; Sivaguru, Barghav S; Sivaguru, Vignesh A; Lu, Xiaochen; Choi, Kyung Hwa; Saif, M Taher A; Lin, Brian; Sadayappan, Sakthivel

    2015-11-01

    The ability to image the entire adult mouse heart at high resolution in 3-D would provide enormous advantages in the study of heart disease. However, a technique for imaging nuclear/cellular detail as well as the overall structure of the entire heart in 3-D with minimal effort is lacking. To solve this problem, we modified the benzyl alcohol:benzyl benzoate (BABB) clearing technique by labeling mouse hearts with periodic acid Schiff (PAS) stain. We then imaged the hearts with a combination of two-photon fluorescence microscopy and automated tile-scan imaging/stitching. Utilizing the differential spectral properties of PAS, we could identify muscle and nuclear compartments in the heart. We were also able to visualize the differences between a 3-month-old normal mouse heart and a mouse heart that had undergone heart failure due to the expression of cardiac myosin binding protein-C (cMyBP-C) gene mutation (t/t). Using 2-D and 3-D morphometric analysis, we found that the t/t heart had anomalous ventricular shape, volume, and wall thickness, as well as a disrupted sarcomere pattern. We further validated our approach using decellularized hearts that had been cultured with 3T3 fibroblasts, which were tracked using a nuclear label. We were able to detect the 3T3 cells inside the decellularized intact heart tissue, achieving nuclear/cellular resolution in 3-D. The combination of labeling, clearing, and two-photon microscopy together with tiling eliminates laborious and time-consuming physical sectioning, alignment, and 3-D reconstruction.

  18. Apertureless near-field/far-field CW two-photon microscope for biological and material imaging and spectroscopic applications.

    Science.gov (United States)

    Nowak, Derek B; Lawrence, A J; Sánchez, Erik J

    2010-12-10

    We present the development of a versatile spectroscopic imaging tool to allow for imaging with single-molecule sensitivity and high spatial resolution. The microscope allows for near-field and subdiffraction-limited far-field imaging by integrating a shear-force microscope on top of a custom inverted microscope design. The instrument has the ability to image in ambient conditions with optical resolutions on the order of tens of nanometers in the near field. A single low-cost computer controls the microscope with a field programmable gate array data acquisition card. High spatial resolution imaging is achieved with an inexpensive CW multiphoton excitation source, using an apertureless probe and simplified optical pathways. The high-resolution, combined with high collection efficiency and single-molecule sensitive optical capabilities of the microscope, are demonstrated with a low-cost CW laser source as well as a mode-locked laser source.

  19. A software solution for recording circadian oscillator features in time-lapse live cell microscopy

    Directory of Open Access Journals (Sweden)

    Salmon Patrick

    2010-07-01

    Full Text Available Abstract Background Fluorescent and bioluminescent time-lapse microscopy approaches have been successfully used to investigate molecular mechanisms underlying the mammalian circadian oscillator at the single cell level. However, most of the available software and common methods based on intensity-threshold segmentation and frame-to-frame tracking are not applicable in these experiments. This is due to cell movement and dramatic changes in the fluorescent/bioluminescent reporter protein during the circadian cycle, with the lowest expression level very close to the background intensity. At present, the standard approach to analyze data sets obtained from time lapse microscopy is either manual tracking or application of generic image-processing software/dedicated tracking software. To our knowledge, these existing software solutions for manual and automatic tracking have strong limitations in tracking individual cells if their plane shifts. Results In an attempt to improve existing methodology of time-lapse tracking of a large number of moving cells, we have developed a semi-automatic software package. It extracts the trajectory of the cells by tracking theirs displacements, makes the delineation of cell nucleus or whole cell, and finally yields measurements of various features, like reporter protein expression level or cell displacement. As an example, we present here single cell circadian pattern and motility analysis of NIH3T3 mouse fibroblasts expressing a fluorescent circadian reporter protein. Using Circadian Gene Express plugin, we performed fast and nonbiased analysis of large fluorescent time lapse microscopy datasets. Conclusions Our software solution, Circadian Gene Express (CGE, is easy to use and allows precise and semi-automatic tracking of moving cells over longer period of time. In spite of significant circadian variations in protein expression with extremely low expression levels at the valley phase, CGE allows accurate and

  20. 3-D time-lapse electrical resistivity monitoring of injected CO2 in a shallow aquifer

    DEFF Research Database (Denmark)

    Doetsch, Joseph A. J.A.; Auken, Esben; Christiansen, Anders Vest C A.V.C.

    2013-01-01

    and 10 m and monitored its migration using 320 surface electrodes on a 126 m × 20 m grid. A fully automated acquisition system continuously collected data and uploaded it into an online database. The large amount of data allows for time-series analysis for data quality and noise estimation. A baseline...... inversion reveals the geology at the site consisting of aeolian sands near the surface and glacial sands below 5 m depth. Time-lapse inversions clearly image the dissolved CO2 plume with decreased electrical resistivity values. We can follow the CO2 plume as it spreads and moves with the groundwater...

  1. A software solution for recording circadian oscillator features in time-lapse live cell microscopy.

    Science.gov (United States)

    Sage, Daniel; Unser, Michael; Salmon, Patrick; Dibner, Charna

    2010-07-06

    Fluorescent and bioluminescent time-lapse microscopy approaches have been successfully used to investigate molecular mechanisms underlying the mammalian circadian oscillator at the single cell level. However, most of the available software and common methods based on intensity-threshold segmentation and frame-to-frame tracking are not applicable in these experiments. This is due to cell movement and dramatic changes in the fluorescent/bioluminescent reporter protein during the circadian cycle, with the lowest expression level very close to the background intensity. At present, the standard approach to analyze data sets obtained from time lapse microscopy is either manual tracking or application of generic image-processing software/dedicated tracking software. To our knowledge, these existing software solutions for manual and automatic tracking have strong limitations in tracking individual cells if their plane shifts. In an attempt to improve existing methodology of time-lapse tracking of a large number of moving cells, we have developed a semi-automatic software package. It extracts the trajectory of the cells by tracking theirs displacements, makes the delineation of cell nucleus or whole cell, and finally yields measurements of various features, like reporter protein expression level or cell displacement. As an example, we present here single cell circadian pattern and motility analysis of NIH3T3 mouse fibroblasts expressing a fluorescent circadian reporter protein. Using Circadian Gene Express plugin, we performed fast and nonbiased analysis of large fluorescent time lapse microscopy datasets. Our software solution, Circadian Gene Express (CGE), is easy to use and allows precise and semi-automatic tracking of moving cells over longer period of time. In spite of significant circadian variations in protein expression with extremely low expression levels at the valley phase, CGE allows accurate and efficient recording of large number of cell parameters, including

  2. Preparation of wholemount mouse intestine for high-resolution three-dimensional imaging using two-photon microscopy.

    Science.gov (United States)

    Appleton, P L; Quyn, A J; Swift, S; Näthke, I

    2009-05-01

    Visualizing overall tissue architecture in three dimensions is fundamental for validating and integrating biochemical, cell biological and visual data from less complex systems such as cultured cells. Here, we describe a method to generate high-resolution three-dimensional image data of intact mouse gut tissue. Regions of highest interest lie between 50 and 200 mum within this tissue. The quality and usefulness of three-dimensional image data of tissue with such depth is limited owing to problems associated with scattered light, photobleaching and spherical aberration. Furthermore, the highest-quality oil-immersion lenses are designed to work at a maximum distance of image at high-resolution deep within tissue. We show that manipulating the refractive index of the mounting media and decreasing sample opacity greatly improves image quality such that the limiting factor for a standard, inverted multi-photon microscope is determined by the working distance of the objective as opposed to detectable fluorescence. This method negates the need for mechanical sectioning of tissue and enables the routine generation of high-quality, quantitative image data that can significantly advance our understanding of tissue architecture and physiology.

  3. Nonlinear spectral imaging of human normal skin, basal cell carcinoma and squamous cell carcinoma based on two-photon excited fluorescence and second-harmonic generation

    Science.gov (United States)

    Xiong, S. Y.; Yang, J. G.; Zhuang, J.

    2011-10-01

    In this work, we use nonlinear spectral imaging based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) for analyzing the morphology of collagen and elastin and their biochemical variations in basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and normal skin tissue. It was found in this work that there existed apparent differences among BCC, SCC and normal skin in terms of their thickness of the keratin and epithelial layers, their size of elastic fibers, as well as their distribution and spectral characteristics of collagen. These differences can potentially be used to distinguish BCC and SCC from normal skin, and to discriminate between BCC and SCC, as well as to evaluate treatment responses.

  4. Dipolar versus octupolar triphenylamine-based fluorescent organic nanoparticles as brilliant one- and two-photon emitters for (bio)imaging.

    Science.gov (United States)

    Parthasarathy, Venkatakrishnan; Fery-Forgues, Suzanne; Campioli, Elisa; Recher, Gaëlle; Terenziani, Francesca; Blanchard-Desce, Mireille

    2011-11-18

    Two related triphenylamine-based dipolar and octupolar fluorophores are used to prepare aqueous suspensions of fluorescent organic nanoparticles (FONs) via the reprecipitation method. The obtained spherical nanoparticles (30-40 nm in diameter) are fluorescent in aqueous solution (up to 15% fluorescence quantum yield) and exhibit extremely high one- and two-photon brightness, superior to those obtained for quantum dots. Despite the two chromophores showing similar fluorescence in solution, the fluorescence of FONs made from the octupolar derivative is significantly red-shifted compared to that generated by the dipolar FONs. In addition, the maximum two-photon absorption cross section of the FONs made from the octupolar derivative is 55% larger than that of the dipolar derivative FONs. The experimental observations provide evidence that the different molecular shape (rodlike versus three-branched) and charge distribution (dipolar versus octupolar) of the two chromophores strongly affect the packing inside the nanoparticles as well as their spectroscopic properties and colloidal stability in pure water. The use of these FONs as probes for biphotonic in-vivo imaging is investigated on Xenopus laevis tadpoles to test their utilization for angiography. When using FONs made from the octupolar dye, the formation of microagglomerates (2-5 μm scale) is observed in vivo, with subsequent lethal occlusion of the blood vessels. Conversely, the nanoparticles of the dipolar dye allow acute imaging of blood vessels thanks to their suitable size and brightness, while no toxic effect is observed. Such a goal cannot be achieved with the dissolved dye, which permeates the vessel walls. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Rock Slope Monitoring from 4D Time-Lapse Structure from Motion Analysis

    Science.gov (United States)

    Kromer, Ryan; Abellan, Antonio; Chyz, Alex; Hutchinson, Jean

    2017-04-01

    more accurate 3D models using time-lapse photos taken from an array of cameras than photos taken from a single camera from multiple positions. For this survey setup, it was possible to detect mm to cm level of changes, which is of sufficient accuracy to detect the pre-failure stage of rockfalls, as well as small rockfall events. Additionally, cameras mounted in a static array can be operated remotely and automatically. Time-lapse SfM photogrammetry can be a cost effective alternative to terrestrial laser scanning for rockfall prone areas and facilitates the study of surface processes with high spatial and temporal detail. We gratefully acknowledge support from the NSERC collaborative research and development grant. References Eltner, A., Kaiser, A., Castillo, C.; Rock, G., Neugirg, F., Abellán, A. Image-based surface reconstruction in geomorphometry—Merits, limits and developments. Earth Surf. Dyn. 2016, 4, 359-389. Kromer, R. A., Abellán, A., Hutchinson, D. J., Lato, M., Edwards, T., & Jaboyedoff, M. A 4D filtering and calibration technique for small-scale point cloud change detection with a terrestrial laser scanner. Remote Sensing 2015, 7(10), 13029-13052.

  6. Two-photon excited fluorescence imaging of the pancreatic solid pseudopapillary tumor without hematoxylin and eosin stains.

    Science.gov (United States)

    Xu, Yahao; Liao, Chenxi; Chen, Jing; Chen, Youting; Zhu, Xiaoqin; Chen, Jianxin

    2016-05-01

    Solid pseudopapillary tumor (SPT) of the pancreas is an epithelial tumor with low-grade malignant potential and present more common in females. At present, the gold standard for accurate diagnosis of pancreatic tumor was mostly depending on the pathological and/or cytological evaluation. In this work, TPEF microscopy was applied to obtain the images of human normal pancreas and SPT of the pancreas without hematoxylin and eosin (H&E) staining, for the purpose of identifying the organization structural, cell morphological, and cytoplasm changing, which were then compared to their corresponding H&E stained histopathological results. Our results showed that high-resolution TPEF imaging of the pancreatic SPT can clearly distinguish the pathological features from normal pancreas in unstained histological sections, and the results are consistent with the histological results. Moreover, we measured the nuclear-cytoplasmic ratios of the pancreatic SPT and normal pancreas to characterize their difference in the cytomorphological feature. It indicated that this technique can achieve the consistent information of pathological diagnosis, and has the potential to substantially improve the optical diagnosis and treatment of the pancreatic SPT without H&E staining in the future. SCANNING 38:245-250, 2016. © 2015 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

  7. Correlating two-photon excited fluorescence imaging of breast cancer cellular redox state with seahorse flux analysis of normalized cellular oxygen consumption

    Science.gov (United States)

    Hou, Jue; Wright, Heather J.; Chan, Nicole; Tran, Richard; Razorenova, Olga V.; Potma, Eric O.; Tromberg, Bruce J.

    2016-06-01

    Two-photon excited fluorescence (TPEF) imaging of the cellular cofactors nicotinamide adenine dinucleotide and oxidized flavin adenine dinucleotide is widely used to measure cellular metabolism, both in normal and pathological cells and tissues. When dual-wavelength excitation is used, ratiometric TPEF imaging of the intrinsic cofactor fluorescence provides a metabolic index of cells-the "optical redox ratio" (ORR). With increased interest in understanding and controlling cellular metabolism in cancer, there is a need to evaluate the performance of ORR in malignant cells. We compare TPEF metabolic imaging with seahorse flux analysis of cellular oxygen consumption in two different breast cancer cell lines (MCF-7 and MDA-MB-231). We monitor metabolic index in living cells under both normal culture conditions and, for MCF-7, in response to cell respiration inhibitors and uncouplers. We observe a significant correlation between the TPEF-derived ORR and the flux analyzer measurements (R=0.7901, p<0.001). Our results confirm that the ORR is a valid dynamic index of cell metabolism under a range of oxygen consumption conditions relevant for cancer imaging.

  8. Opposite reactivity of meningeal versus cortical microvessels to the nitric oxide donor glyceryl trinitrate evaluated in vivo with two-photon imaging.

    Directory of Open Access Journals (Sweden)

    Evgeny Pryazhnikov

    Full Text Available Vascular changes underlying headache in migraine patients induced by Glyceryl trinitrate (GTN were previously studied with various imaging techniques. Despite the long history of medical and experimental use of GTN, its effects on the brain vasculature are still poorly understood presumably due to low spatial resolution of the imaging modalities used so far. We took advantage of the micrometer-scale vertical resolution of two-photon microscopy to differentiate between the vasodynamic effects of GTN on meningeal versus cortical vessels imaged simultaneously in anesthetized rats through either thinned skull or glass-sealed cranial window. Intermediate and small calibre vessels were visualized in vivo by imaging intravascular fluorescent dextran, and detection of blood flow direction allowed identification of individual arterioles and venules. We found that i.p.-injected GTN induced a transient constriction of meningeal arterioles, while their cortical counterparts were, in contrast, dilated. These opposing effects of GTN were restricted to arterioles, whereas the effects on venules were insignificant. Interestingly, the NO synthase inhibitor L-NAME did not affect the diameter of meningeal vessels but induced a constriction of cortical vessels. The different cellular environment in cortex versus meninges as well as distinct vessel wall anatomical features probably play crucial role in the observed phenomena. These findings highlight differential region- and vessel-type-specific effects of GTN on cranial vessels, and may implicate new vascular mechanisms of NO-mediated primary headaches.

  9. Calibrating Vadose Zone Models with Time-Lapse Gravity Data

    DEFF Research Database (Denmark)

    Christiansen, Lars; Hansen, A. B.; Looms, M. C.

    2009-01-01

    A change in soil water content is a change in mass stored in the subsurface. Given that the mass change is big enough, the change can be measured with a gravity meter. Attempts have been made with varying success over the last decades to use ground-based time-lapse gravity measurements to infer...... hydrogeological parameters. These studies focused on the saturated zone with specific yield as the most prominent target parameter. Any change in storage in the vadose zone has been considered as noise. Our modeling results show a measureable change in gravity from the vadose zone during a forced infiltration...... experiment on 10m by 10m grass land. Simulation studies show a potential for vadose zone model calibration using gravity data in conjunction with other geophysical data, e.g. cross-borehole georadar. We present early field data and calibration results from a forced infiltration experiment conducted over 30...

  10. Combined time-lapse cinematography and immuno-electron microscopy.

    Science.gov (United States)

    Balfour, B M; Goscicka, T; MacKenzie, J L; Gautam, A; Tate, M; Clark, J

    1990-04-01

    A method was developed to record interactions between mobile non-adherent immunocytes by time-lapse cinematography and then to study the same cells by immuno-electron microscopy, using monoclonal antibodies against surface components. For this purpose a modified stage was designed to fit an inverted microscope. The attachment included a device to cool the culture chamber with N2 gas, a micro-injector for monoclonal antibody and immuno-gold treatment, and two pairs of washing needles to change the medium without disturbance. The technique was first employed to study the formation of aggregates around the antigen-presenting cells in cultures containing cells from hyper-immunized animals. Recently peripheral blood cells from normal subjects and patients with immune deficiency syndromes were stimulated with pokeweed mitogen, cluster formation was recorded, and the cells were processed for immuno-electron microscopy.

  11. A Prototype System for Time-Lapse Electrical Resistivity Tomographies

    Directory of Open Access Journals (Sweden)

    Raffaele Luongo

    2012-01-01

    Full Text Available A prototype system for time-lapse acquisition of 2D electrical resistivity tomography (ERT and time domain reflectometry (TDR measurements was installed in a test site affected by a landslide in Basilicata region (southern Italy. The aim of the system is to monitor in real-time the rainwater infiltration into the soil and obtain information about the variation of the water content in the first layers of the subsoil and the possible influence of this variation on landslide activity. A rain gauge placed in the test site gives information on the rainfall intensity and frequency and suggests the acquisition time interval. The installed system and the preliminary results are presented in this paper.

  12. Model-Based Generation of Synthetic 3D Time-Lapse Sequences of Motile Cells with Growing Filopodia

    OpenAIRE

    Sorokin , Dmitry ,; Peterlik , Igor; Ulman , Vladimír ,; Svoboda , David; Maška , Martin

    2017-01-01

    International audience; The existence of benchmark datasets is essential to objectively evaluate various image analysis methods. Nevertheless, manual annotations of fluorescence microscopy image data are very laborious and not often practicable, especially in the case of 3D+t experiments. In this work, we propose a simulation system capable of generating 3D time-lapse sequences of single motile cells with filopodial protrusions, accompanied by inherently generated ground truth. The system con...

  13. Intravital two-photon microscopy of immune cell dynamics in corneal lymphatic vessels.

    Directory of Open Access Journals (Sweden)

    Philipp Steven

    Full Text Available BACKGROUND: The role of lymphatic vessels in tissue and organ transplantation as well as in tumor growth and metastasis has drawn great attention in recent years. METHODOLOGY/PRINCIPAL FINDINGS: We now developed a novel method using non-invasive two-photon microscopy to simultaneously visualize and track specifically stained lymphatic vessels and autofluorescent adjacent tissues such as collagen fibrils, blood vessels and immune cells in the mouse model of corneal neovascularization in vivo. The mouse cornea serves as an ideal tissue for this technique due to its easy accessibility and its inducible and modifiable state of pathological hem- and lymphvascularization. Neovascularization was induced by suture placement in corneas of Balb/C mice. Two weeks after treatment, lymphatic vessels were stained intravital by intrastromal injection of a fluorescently labeled LYVE-1 antibody and the corneas were evaluated in vivo by two-photon microscopy (TPM. Intravital TPM was performed at 710 nm and 826 nm excitation wavelengths to detect immunofluorescence and tissue autofluorescence using a custom made animal holder. Corneas were then harvested, fixed and analyzed by histology. Time lapse imaging demonstrated the first in vivo evidence of immune cell migration into lymphatic vessels and luminal transport of individual cells. Cells immigrated within 1-5.5 min into the vessel lumen. Mean velocities of intrastromal corneal immune cells were around 9 µm/min and therefore comparable to those of T-cells and macrophages in other mucosal surfaces. CONCLUSIONS: To our knowledge we here demonstrate for the first time the intravital real-time transmigration of immune cells into lymphatic vessels. Overall this study demonstrates the valuable use of intravital autofluorescence two-photon microscopy in the model of suture-induced corneal vascularizations to study interactions of immune and subsequently tumor cells with lymphatic vessels under close as possible

  14. Automated filtering of intrinsic movement artifacts during two-photon intravital microscopy.

    Directory of Open Access Journals (Sweden)

    Denis Soulet

    Full Text Available In vivo imaging using two-photon microscopy is an essential tool to explore the dynamic of physiological events deep within biological tissues for short or extended periods of time. The new capabilities offered by this technology (e.g. high tissue penetrance, low toxicity have opened a whole new era of investigations in modern biomedical research. However, the potential of using this promising technique in tissues of living animals is greatly limited by the intrinsic irregular movements that are caused by cardiac and respiratory cycles and muscular and vascular tone. Here, we show real-time imaging of the brain, spinal cord, sciatic nerve and myenteric plexus of living mice using a new automated program, named Intravital_Microscopy_Toolbox, that removes frames corrupted with motion artifacts from time-lapse videos. Our approach involves generating a dissimilarity score against precalculated reference frames in a specific reference channel, thus allowing the gating of distorted, out-of-focus or translated frames. Since the algorithm detects the uneven peaks of image distortion caused by irregular animal movements, the macro allows a fast and efficient filtering of the image sequence. In addition, extra features have been implemented in the macro, such as XY registration, channel subtraction, extended field of view with maximum intensity projection, noise reduction with average intensity projections, and automated timestamp and scale bar overlay. Thus, the Intravital_Microscopy_Toolbox macro for ImageJ provides convenient tools for biologists who are performing in vivo two-photon imaging in tissues prone to motion artifacts.

  15. Embryo selection: the role of time-lapse monitoring.

    Science.gov (United States)

    Kovacs, Peter

    2014-12-15

    In vitro fertilization has been available for over 3 decades. Its use is becoming more widespread worldwide, and in the developed world, up to 5% of children have been born following IVF. It is estimated that over 5 million children have been conceived in vitro. In addition to giving hope to infertile couples to have their own family, in vitro fertilization has also introduced risks as well. The risk of multiple gestation and the associated maternal and neonatal morbidity/mortality has increased significantly over the past few decades. While stricter transfer policies have eliminated the majority of the high-order multiples, these changes have not yet had much of an impact on the incidence of twins. A twin pregnancy can be avoided by the transfer of a single embryo only. However, the traditionally used method of morphologic embryo selection is not predictive enough to allow routine single embryo transfer; therefore, new screening tools are needed. Time-lapse embryo monitoring allows continuous, non-invasive embryo observation without the need to remove the embryo from optimal culturing conditions. The extra information on the cleavage pattern, morphologic changes and embryo development dynamics could help us identify embryos with a higher implantation potential. These technologic improvements enable us to objectively select the embryo(s) for transfer based on certain algorithms. In the past 5-6 years, numerous studies have been published that confirmed the safety of time-lapse technology. In addition, various markers have already been identified that are associated with the minimal likelihood of implantation and others that are predictive of blastocyst development, implantation potential, genetic health and pregnancy. Various groups have proposed different algorithms for embryo selection based on mostly retrospective data analysis. However, large prospective trials are needed to study the full benefit of these (and potentially new) algorithms before their

  16. Possible mechanism of polyspermy block in human oocytes observed by time-lapse cinematography.

    Science.gov (United States)

    Mio, Yasuyuki; Iwata, Kyoko; Yumoto, Keitaro; Kai, Yoshiteru; Sargant, Haruka C; Mizoguchi, Chizuru; Ueda, Minako; Tsuchie, Yuka; Imajo, Akifumi; Iba, Yumiko; Nishikori, Kyoko

    2012-09-01

    To analyze the fertilization process related to polyspermy block in human oocytes using an in vitro culturing system for time-lapse cinematography. We had 122 oocytes donated for this study from couples that provided informed consent. We recorded human oocytes at 2,000 to 2,800 frames every 10 s during the fertilization process and thereafter every 2 min using a new in vitro culture system originally developed by the authors for time-lapse cinematography. We displayed 30 frames per second for analysis of the polyspermy block during fertilization. Three oocytes showed the leading and following sperm within the zona pellucida in the same microscopic field. The dynamic images obtained during the fertilization process using this new system revealed that once a leading sperm penetrated the zona pellucida and attached to the oocyte membrane, a following sperm was arrested from further penetration into the zona pellucida within 10 s. The present results strongly suggest the existence of a novel mechanism of polyspermy block that takes place at the zona pellucida immediately after fertilization. These findings are clearly different from previous mechanisms describing polyspermy block as the oocyte membrane block to sperm penetration and the zona reaction. The finding presented herein thus represents a novel discovery about the highly complicated polyspermy block mechanism occurring in human oocytes.

  17. Time-lapse cinematography in living Drosophila tissues: preparation of material.

    Science.gov (United States)

    Davis, Ilan; Parton, Richard M

    2006-11-01

    The fruit fly, Drosophila melanogaster, has been an extraordinarily successful model organism for studying the genetic basis of development and evolution. It is arguably the best-understood complex multicellular model system, owing its success to many factors. Recent developments in imaging techniques, in particular sophisticated fluorescence microscopy methods and equipment, now allow cellular events to be studied at high resolution in living material. This ability has enabled the study of features that tend to be lost or damaged by fixation, such as transient or dynamic events. Although many of the techniques of live cell imaging in Drosophila are shared with the greater community of cell biologists working on other model systems, studying living fly tissues presents unique difficulties in keeping the cells alive, introducing fluorescent probes, and imaging through thick hazy cytoplasm. This protocol outlines the preparation of major tissue types amenable to study by time-lapse cinematography and different methods for keeping them alive.

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

    Science.gov (United States)

    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.

  19. Interface-Targeting Strategy Enables Two-Photon Fluorescent Lipid Droplet Probes for High-Fidelity Imaging of Turbid Tissues and Detecting Fatty Liver.

    Science.gov (United States)

    Guo, Lifang; Tian, Minggang; Feng, Ruiqing; Zhang, Ge; Zhang, Ruoyao; Li, Xuechen; Liu, Zhiqiang; He, Xiuquan; Sun, Jing Zhi; Yu, Xiaoqiang

    2018-04-04

    Lipid droplets (LDs) with unique interfacial architecture not only play crucial roles in protecting a cell from lipotoxicity and lipoapoptosis but also closely relate with many diseases such as fatty liver and diabetes. Thus, as one of the important applied biomaterials, fluorescent probes with ultrahigh selectivity for in situ and high-fidelity imaging of LDs in living cells and tissues are critical to elucidate relevant physiological and pathological events as well as detect related diseases. However, available probes only utilizing LDs' waterless neutral cores but ignoring the unique phospholipid monolayer interfaces exhibit low selectivity. They cannot differentiate neutral cores of LDs from intracellular other lipophilic microenvironments, which results in extensively cloud-like background noise and severely limited their bioapplications. Herein, to design LD probes with ultrahigh selectivity, the exceptional interfacial architecture of LDs is considered adequately and thus an interface-targeting strategy is proposed for the first time. According to the novel strategy, we have developed two amphipathic fluorescent probes (N-Cy and N-Py) by introducing different cations into a lipophilic fluorophore (nitrobenzoxadiazole (NBD)). Consequently, their cationic moiety precisely locates the interfaces through electrostatic interaction and simultaneously NBD entirely embeds into the waterless core via hydrophobic interaction. Thus, high-fidelity and background-free fluorescence imaging of LDs are expectably realized in living cells in situ. Moreover, LDs in turbid tissues like skeletal muscle slices have been clearly imaged (up to 82 μm depth) by a two-photon microscope. Importantly, using N-Cy, we not only intuitively monitored the variations of LDs in number, size, and morphology but also clearly revealed their abnormity in hepatic tissues resulting from fatty liver. Therefore, these unique probes provide excellent imaging tools for elucidating LD

  20. Three-dimensional cell organization leads to almost immediate HRE activity as demonstrated by molecular imaging of MG-63 spheroids using two-photon excitation microscopy.

    Science.gov (United States)

    Indovina, Paola; Collini, Maddalena; Chirico, Giuseppe; Santini, Maria Teresa

    2007-02-20

    Hypoxia through HRE (hypoxia-responsive element) activity in MG-63 human osteosarcoma cells grown in monolayer and as very small, three-dimensional tumor spheroids was investigated using molecular imaging techniques. MG-63 cells were stably transfected with a vector constructed with multiple copies of the HRE sequence of the human vascular endothelial growth factor (VEGF) gene and with the enhanced green fluorescent protein (EGFP) coding sequence. During hypoxia when HIF-1alpha (hypoxia-inducible factor-1alpha) is stabilized, the binding of HIF-1 to the HRE sequences of the vector allows the transcription of EGFP and the appearance of fluorescence. Transfected monolayer cells were characterized by flow cytometric analysis in response to various hypoxic conditions and HIF-1alpha expression in these cells was assessed by Western blotting. Two-photon excitation (TPE) microscopy was then used to examine both MG-63-transfected monolayer cells and spheroids at 2 and 5 days of growth in normoxic conditions. Monolayer cells reveal almost no fluorescence, whereas even very small spheroids (HRE activation. This activation of the HRE sequences, which control a wide variety of genes, suggests that monolayer cells and spheroids of the MG-63 cell line have different genes activated and thus diverse functional activities.

  1. Network Analysis of Time-Lapse Microscopy Recordings

    Directory of Open Access Journals (Sweden)

    Erik eSmedler

    2014-09-01

    Full Text Available Multicellular organisms rely on intercellular communication to regulate important cellular processes critical to life. To further our understanding of those processes there is a need to scrutinize dynamical signaling events and their functions in both cells and organisms. Here, we report a method and provide MATLAB code that analyzes time-lapse microscopy recordings to identify and characterize network structures within large cell populations, such as interconnected neurons. The approach is demonstrated using intracellular calcium (Ca2+ recordings in neural progenitors and cardiac myocytes, but could be applied to a wide variety of biosensors employed in diverse cell types and organisms. In this method, network structures are analyzed by applying cross-correlation signal processing and graph theory to single-cell recordings. The goal of the analysis is to determine if the single cell activity constitutes a network of interconnected cells and to decipher the properties of this network. The method can be applied in many fields of biology in which biosensors are used to monitor signaling events in living cells. Analyzing intercellular communication in cell ensembles can reveal essential network structures that provide important biological insights.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

  5. Two-Photon Processor and SeNeCA: a freely available software package to process data from two-photon calcium imaging at speeds down to several milliseconds per frame

    Czech Academy of Sciences Publication Activity Database

    Tomek, J.; Novák, Ondřej; Syka, Josef

    2013-01-01

    Roč. 110, č. 1 (2013), s. 243-256 ISSN 0022-3077 R&D Projects: GA ČR(CZ) GAP303/12/1347; GA ČR(CZ) GBP304/12/G069 Institutional support: RVO:68378041 Keywords : algorithm * segmentation * calcium imaging Subject RIV: FH - Neurology Impact factor: 3.041, year: 2013

  6. On the possibility of time-lapse ultrahigh-resolution optical coherence tomography for bladder cancer grading

    Science.gov (United States)

    Yuan, Zhijia; Chen, Bai; Ren, Hugang; Pan, Yingtian

    2009-09-01

    It has been recently demonstrated that the cellular details of bladder epithelium embedded in speckle noise can be uncovered with time-lapse ultrahigh-resolution optical coherence tomography (TL-uOCT) by proper time-lapse frame averaging that takes advantage of cellular micromotion in fresh biological tissue ex vivo. Here, spectral-domain 3-D TL-uOCT is reported to further improve the image fidelity, and new experimental evidence is presented to differentiate normal and cancerous nuclei of rodent bladder epithelia. Results of animal cancer study reveal that despite a slight overestimation (e.g., cancerous (e.g., high-grade DN''~13 μm) urothelia, which may potentially be very useful for enhancing the diagnosis of nonpapillary bladder cancer. More animal study is being conducted to examine the utility to differentiate hyperplasia, dysplasia, and carcinoma in situ.

  7. Two-photon microscopy imaging of thy1GFP-M transgenic mice: a novel animal model to investigate brain dendritic cell subsets in vivo.

    Directory of Open Access Journals (Sweden)

    Claudia Laperchia

    Full Text Available Transgenic mice expressing fluorescent proteins in specific cell populations are widely used for in vivo brain studies with two-photon fluorescence (TPF microscopy. Mice of the thy1GFP-M line have been engineered for selective expression of green fluorescent protein (GFP in neuronal populations. Here, we report that TPF microscopy reveals, at the brain surface of these mice, also motile non-neuronal GFP+ cells. We have analyzed the behavior of these cells in vivo and characterized in brain sections their immunophenotype.With TPF imaging, motile GFP+ cells were found in the meninges, subarachnoid space and upper cortical layers. The striking feature of these cells was their ability to move across the brain parenchyma, exhibiting evident shape changes during their scanning-like motion. In brain sections, GFP+ cells were immunonegative to antigens recognizing motile cells such as migratory neuroblasts, neuronal and glial precursors, mast cells, and fibroblasts. GFP+ non-neuronal cells exhibited instead the characteristic features and immunophenotype (CD11c and major histocompatibility complex molecule class II immunopositivity of dendritic cells (DCs, and were immunonegative to the microglial marker Iba-1. GFP+ cells were also identified in lymph nodes and blood of thy1GFP-M mice, supporting their identity as DCs. Thus, TPF microscopy has here allowed the visualization for the first time of the motile behavior of brain DCs in situ. The results indicate that the thy1GFP-M mouse line provides a novel animal model for the study of subsets of these professional antigen-presenting cells in the brain. Information on brain DCs is still very limited and imaging in thy1GFP-M mice has a great potential for analyses of DC-neuron interaction in normal and pathological conditions.

  8. Exploring Time-Lapse Photography as a Means for Qualitative Data Collection

    Science.gov (United States)

    Persohn, Lindsay

    2015-01-01

    Collecting information via time-lapse photography is nothing new. Scientists and artists have been using this kind of data since the late 1800s. However, my research and experiments with time-lapse have shown that great potential may lie in its application to educational and social scientific research methods. This article is part history, part…

  9. Quantifying Effusion Rates at Active Volcanoes through Integrated Time-Lapse Laser Scanning and Photography

    Directory of Open Access Journals (Sweden)

    Neil Slatcher

    2015-11-01

    Full Text Available During volcanic eruptions, measurements of the rate at which magma is erupted underpin hazard assessments. For eruptions dominated by the effusion of lava, estimates are often made using satellite data; here, in a case study at Mount Etna (Sicily, we make the first measurements based on terrestrial laser scanning (TLS, and we also include explosive products. During the study period (17–21 July 2012, regular Strombolian explosions were occurring within the Bocca Nuova crater, producing a ~50 m-high scoria cone and a small lava flow field. TLS surveys over multi-day intervals determined a mean cone growth rate (effusive and explosive products of ~0.24 m3·s−1. Differences between 0.3-m resolution DEMs acquired at 10-minute intervals captured the evolution of a breakout lava flow lobe advancing at 0.01–0.03 m3·s−1. Partial occlusion within the crater prevented similar measurement of the main flow, but integrating TLS data with time-lapse imagery enabled lava viscosity (7.4 × 105 Pa·s to be derived from surface velocities and, hence, a flux of 0.11 m3·s−1 to be calculated. Total dense rock equivalent magma discharge estimates are ~0.1–0.2 m3·s−1 over the measurement period and suggest that simultaneous estimates from satellite data are somewhat overestimated. Our results support the use of integrated TLS and time-lapse photography for ground-truthing space-based measurements and highlight the value of interactive image analysis when automated approaches, such as particle image velocimetry (PIV, fail.

  10. Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy.

    Directory of Open Access Journals (Sweden)

    Masahito Asada

    Full Text Available BACKGROUND: Babesia bovis is an apicomplexan intraerythrocytic protozoan parasite that induces babesiosis in cattle after transmission by ticks. During specific stages of the apicomplexan parasite lifecycle, such as the sporozoites of Plasmodium falciparum and tachyzoites of Toxoplasma gondii, host cells are targeted for invasion using a unique, active process termed "gliding motility". However, it is not thoroughly understood how the merozoites of B. bovis target and invade host red blood cells (RBCs, and gliding motility has so far not been observed in the parasite. METHODOLOGY/PRINCIPAL FINDINGS: Gliding motility of B. bovis merozoites was revealed by time-lapse video microscopy. The recorded images revealed that the process included egress of the merozoites from the infected RBC, gliding motility, and subsequent invasion into new RBCs. The gliding motility of B. bovis merozoites was similar to the helical gliding of Toxoplasma tachyzoites. The trails left by the merozoites were detected by indirect immunofluorescence assay using antiserum against B. bovis merozoite surface antigen 1. Inhibition of gliding motility by actin filament polymerization or depolymerization indicated that the gliding motility was driven by actomyosin dependent process. In addition, we revealed the timing of breakdown of the parasitophorous vacuole. Time-lapse image analysis of membrane-stained bovine RBCs showed formation and breakdown of the parasitophorous vacuole within ten minutes of invasion. CONCLUSIONS/SIGNIFICANCE: This is the first report of the gliding motility of B. bovis. Since merozoites of Plasmodium parasites do not glide on a substrate, the gliding motility of B. bovis merozoites is a notable finding.

  11. Active and passive electrical and seismic time-lapse monitoring of earthen embankments

    Science.gov (United States)

    Rittgers, Justin Bradley

    In this dissertation, I present research involving the application of active and passive geophysical data collection, data assimilation, and inverse modeling for the purpose of earthen embankment infrastructure assessment. Throughout the dissertation, I identify several data characteristics, and several challenges intrinsic to characterization and imaging of earthen embankments and anomalous seepage phenomena, from both a static and time-lapse geophysical monitoring perspective. I begin with the presentation of a field study conducted on a seeping earthen dam, involving static and independent inversions of active tomography data sets, and self-potential modeling of fluid flow within a confined aquifer. Additionally, I present results of active and passive time-lapse geophysical monitoring conducted during two meso-scale laboratory experiments involving the failure and self-healing of embankment filter materials via induced vertical cracking. Identified data signatures and trends, as well as 4D inversion results, are discussed as an underlying motivation for conducting subsequent research. Next, I present a new 4D acoustic emissions source localization algorithm that is applied to passive seismic monitoring data collected during a full-scale embankment failure test. Acoustic emissions localization results are then used to help spatially constrain 4D inversion of collocated self-potential monitoring data. I then turn to time-lapse joint inversion of active tomographic data sets applied to the characterization and monitoring of earthen embankments. Here, I develop a new technique for applying spatiotemporally varying structural joint inversion constraints. The new technique, referred to as Automatic Joint Constraints (AJC), is first demonstrated on a synthetic 2D joint model space, and is then applied to real geophysical monitoring data sets collected during a full-scale earthen embankment piping-failure test. Finally, I discuss some non-technical issues related to

  12. Time-lapse three-dimensional inversion of complex conductivity data using an active time constrained (ATC) approach

    Science.gov (United States)

    Karaoulis, M.; Revil, A.; Werkema, D.D.; Minsley, B.J.; Woodruff, W.F.; Kemna, A.

    2011-01-01

    Induced polarization (more precisely the magnitude and phase of impedance of the subsurface) is measured using a network of electrodes located at the ground surface or in boreholes. This method yields important information related to the distribution of permeability and contaminants in the shallow subsurface. We propose a new time-lapse 3-D modelling and inversion algorithm to image the evolution of complex conductivity over time. We discretize the subsurface using hexahedron cells. Each cell is assigned a complex resistivity or conductivity value. Using the finite-element approach, we model the in-phase and out-of-phase (quadrature) electrical potentials on the 3-D grid, which are then transformed into apparent complex resistivity. Inhomogeneous Dirichlet boundary conditions are used at the boundary of the domain. The calculation of the Jacobian matrix is based on the principles of reciprocity. The goal of time-lapse inversion is to determine the change in the complex resistivity of each cell of the spatial grid as a function of time. Each model along the time axis is called a 'reference space model'. This approach can be simplified into an inverse problem looking for the optimum of several reference space models using the approximation that the material properties vary linearly in time between two subsequent reference models. Regularizations in both space domain and time domain reduce inversion artefacts and improve the stability of the inversion problem. In addition, the use of the time-lapse equations allows the simultaneous inversion of data obtained at different times in just one inversion step (4-D inversion). The advantages of this new inversion algorithm are demonstrated on synthetic time-lapse data resulting from the simulation of a salt tracer test in a heterogeneous random material described by an anisotropic semi-variogram. ?? 2011 The Authors Geophysical Journal International ?? 2011 RAS.

  13. A Versatile Time-Lapse Camera System Developed by the Hawaiian Volcano Observatory for Use at Kilauea Volcano, Hawaii

    Science.gov (United States)

    Orr, Tim R.; Hoblitt, Richard P.

    2008-01-01

    Volcanoes can be difficult to study up close. Because it may be days, weeks, or even years between important events, direct observation is often impractical. In addition, volcanoes are often inaccessible due to their remote location and (or) harsh environmental conditions. An eruption adds another level of complexity to what already may be a difficult and dangerous situation. For these reasons, scientists at the U.S. Geological Survey (USGS) Hawaiian Volcano Observatory (HVO) have, for years, built camera systems to act as surrogate eyes. With the recent advances in digital-camera technology, these eyes are rapidly improving. One type of photographic monitoring involves the use of near-real-time network-enabled cameras installed at permanent sites (Hoblitt and others, in press). Time-lapse camera-systems, on the other hand, provide an inexpensive, easily transportable monitoring option that offers more versatility in site location. While time-lapse systems lack near-real-time capability, they provide higher image resolution and can be rapidly deployed in areas where the use of sophisticated telemetry required by the networked cameras systems is not practical. This report describes the latest generation (as of 2008) time-lapse camera system used by HVO for photograph acquisition in remote and hazardous sites on Kilauea Volcano.

  14. Preparation of a Two-Photon Fluorescent Probe for Imaging H2O2 in Lysosomes in Living Cells and Tissues.

    Science.gov (United States)

    Ren, Mingguang; Deng, Beibei; Kong, Xiuqi; Tang, Yonghe; Lin, Weiying

    2017-01-01

    Hydrogen peroxide (H 2 O 2 ) plays important roles in many physiological and pathological processes. At the cellular organelle level, the abnormal concentrations of H 2 O 2 in the lysosomes may cause redox imbalance and the loss of the critical functions of the lysosomes. Herein, we describe the preparation of a potent lysosome-targeted two-photon fluorescent probe (Lyso-HP) for the detection of H 2 O 2 in the lysosomes in the living cells. This unique fluorescent probe can also be employed to effectively detect H 2 O 2 in the living tissues using two-photon fluorescence microscopy.

  15. 3D time-lapse analysis of Rab11/FIP5 complex: spatiotemporal dynamics during apical lumen formation.

    Science.gov (United States)

    Mangan, Anthony; Prekeris, Rytis

    2015-01-01

    Fluorescent imaging of fixed cells grown in two-dimensional (2D) cultures is one of the most widely used techniques for observing protein localization and distribution within cells. Although this technique can also be applied to polarized epithelial cells that form three-dimensional (3D) cysts when grown in a Matrigel matrix suspension, there are still significant limitations in imaging cells fixed at a particular point in time. Here, we describe the use of 3D time-lapse imaging of live cells to observe the dynamics of apical membrane initiation site (AMIS) formation and lumen expansion in polarized epithelial cells.

  16. Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring

    DEFF Research Database (Denmark)

    Ingerslev, Hans Jakob; Hindkjær, Johnny Juhl; Kirkegaard, Kirstine

    2012-01-01

    recently demonstrated to occur from first cleavage cycle in mice using time-lapse microscopy, with the largest impact on the pre-compaction stages. However, embryonic development in mice differs in many aspects from human embryonic development. The objective of this retrospective, descriptive study...... was to evaluate the influence of oxygen tension on human pre-implantation development using time-lapse monitoring. Materials and methods: Human embryos were cultured to the blastocyst stage in a time-lapse incubator (EmbryoScope™) in 20% O2 (group 1), 20% O2 for 24 hours followed by culture in 5% O2 (group 2......) or in 5% O2 (group 3). Eligible were patients with age 8 oocytes retrieved. Group 1 consisted of 120 IVF/ICSI embryos from 26 patients recruited to a study conducted to evaluate the safety of the time-lapse incubator by randomising 1:1 embryos from a patient to culture...

  17. History matching of time-lapse crosswell data using ensemble kalman filtering

    KAUST Repository

    de Matos Ravanelli, Fabio Miguel

    2015-09-03

    Data from crosswell seismic surveys is processed to provide crosswell time-lapse data to map fluid changes in a reservoir where time-lapse or 4D seismic data is unavailable or unreliable, such as in onshore reservoirs. The resultant processing results provide quantitative information for history matching purposes using a probabilistic approach to take in account uncertainties in the geological model and reduce uncertainties in reservoir production forecasts.

  18. MitoGen: A Framework for Generating 3D Synthetic Time-Lapse Sequences of Cell Populations in Fluorescence Microscopy.

    Science.gov (United States)

    Svoboda, David; Ulman, Vladimir

    2017-01-01

    The proper analysis of biological microscopy images is an important and complex task. Therefore, it requires verification of all steps involved in the process, including image segmentation and tracking algorithms. It is generally better to verify algorithms with computer-generated ground truth datasets, which, compared to manually annotated data, nowadays have reached high quality and can be produced in large quantities even for 3D time-lapse image sequences. Here, we propose a novel framework, called MitoGen, which is capable of generating ground truth datasets with fully 3D time-lapse sequences of synthetic fluorescence-stained cell populations. MitoGen shows biologically justified cell motility, shape and texture changes as well as cell divisions. Standard fluorescence microscopy phenomena such as photobleaching, blur with real point spread function (PSF), and several types of noise, are simulated to obtain realistic images. The MitoGen framework is scalable in both space and time. MitoGen generates visually plausible data that shows good agreement with real data in terms of image descriptors and mean square displacement (MSD) trajectory analysis. Additionally, it is also shown in this paper that four publicly available segmentation and tracking algorithms exhibit similar performance on both real and MitoGen-generated data. The implementation of MitoGen is freely available.

  19. Monotoring of CO2 Sequestration at Sleipner Using Full Waveform Inversion in Time-lapse Mode.

    Science.gov (United States)

    Gosselet, A.; Singh, S. C.

    2007-12-01

    It is now widely admitted that recent increase of CO2 in the atmosphere is due to human activities. The consecutive greenhouse effect is a major ecological concern. Geological storage is one proposed way to reduce atmosphere CO2 emissions. The Sleipner methane field, North Sea, is the very first site where CO2 has been injected back into a deep saline aquifer. In 1996, the Norwegian company Statoil and its partners began the production of the methane. The extracted methane contains a relatively high ratio of CO2, between 4% and 9%, that has to be reduced below 2.5% before delivering into the pipeline. An environmental tax introduced in Norway as early as 1991 prompted the company to store the separated CO2 instead of releasing it into the atmosphere as usually done. The CO2 is injected at the base of the Utsira sands. This water bearing formation lies at a depth between 800 and 1000m and is sealed by a thick shale layer. Seismic monitoring is a key tool in this strategy from a security standpoint and for sequestration optimization itself. Consequently, 3D seismic data were acquired before injection in 1994 and after injection in 1999, 2001, 2002, 2004 and 2006. Well-log revealed that the reservoir is crossed by thin shale layers that are 1 to 10m thick. CO2 rises up and is confined vertically by the shale layers, favouring horizontal gas migration and creating gas bearing thin beds. Seismic imaging of the gas pockets is therefore a challenging problem because large velocity variations occur on very short distance. Classical processing of time-lapse data consists in subtracting repeated survey seismic traces from the pre- injection baseline traces to exhibit changes within the reservoir. This approach remains qualitative, providing only the shape and extent of the gas cloud. Instead, we propose to compare elastic models of the subsurface computed through 2D full wave form inversion, an advanced seismic imaging technique. This method is based on the wave equation

  20. ESIAC: A data products system for ERTS imagery (time-lapse viewing and measuring)

    Science.gov (United States)

    Evans, W. E.; Serebreny, S. M.

    1974-01-01

    An Electronic Satellite Image Analysis Console (ESIAC) has been developed for visual analysis and objective measurement of earth resources imagery. The system is being employed to process imagery for use by USGS investigators in several different disciplines studying dynamic hydrologic conditions. The ESIAC provides facilities for storing registered image sequences in a magnetic video disc memory for subsequent recall, enhancement, and animated display in monochrome or color. The unique feature of the system is the capability to time-lapse the ERTS imagery and/or analytic displays of the imagery. Data products have included quantitative measurements of distances and areas, brightness profiles, and movie loops of selected themes. The applications of these data products are identified and include such diverse problem areas as measurement of snowfield extent, sediment plumes from estuary dicharge, playa inventory, phreatophyte and other vegetation changes. A comparative ranking of the electronic system in terms of accuracy, cost effectiveness and data output shows it to be a viable means of data analysis.

  1. Time-lapse analysis of methane quantity in Mary Lee group of coal seams using filter-based multiple-point geostatistical simulation

    Science.gov (United States)

    Karacan, C. Özgen; Olea, Ricardo A.

    2013-01-01

    Coal seam degasification and its success are important for controlling methane, and thus for the health and safety of coal miners. During the course of degasification, properties of coal seams change. Thus, the changes in coal reservoir conditions and in-place gas content as well as methane emission potential into mines should be evaluated by examining time-dependent changes and the presence of major heterogeneities and geological discontinuities in the field. In this work, time-lapsed reservoir and fluid storage properties of the New Castle coal seam, Mary Lee/Blue Creek seam, and Jagger seam of Black Warrior Basin, Alabama, were determined from gas and water production history matching and production forecasting of vertical degasification wellbores. These properties were combined with isotherm and other important data to compute gas-in-place (GIP) and its change with time at borehole locations. Time-lapsed training images (TIs) of GIP and GIP difference corresponding to each coal and date were generated by using these point-wise data and Voronoi decomposition on the TI grid, which included faults as discontinuities for expansion of Voronoi regions. Filter-based multiple-point geostatistical simulations, which were preferred in this study due to anisotropies and discontinuities in the area, were used to predict time-lapsed GIP distributions within the study area. Performed simulations were used for mapping spatial time-lapsed methane quantities as well as their uncertainties within the study area.

  2. In-situ, time-lapse study of extracellular polymeric substance discharge in Streptococcus mutans biofilm.

    Science.gov (United States)

    Liu, Bernard Haochih; Yu, Li-Chieh

    2017-02-01

    Streptococcus mutans is one of the main pathogens that cause tooth decay. By metabolizing carbohydrates, S. mutans emits extracellular polymeric substance (EPS) that adheres to the tooth surface and forms layers of biofilm. Periodontal disease occurs due to the low pH environment created by S. mutans biofilm, and such an acidic environment gradually erodes tooth enamel. Since the existence of EPS is essential in the formation of biofilm, the in-situ investigation of its generation and distribution in real time is the key to the control and suppression of S. mutans biofilm. Prior studies of the biofilm formation process by fluorescence microscope, scanning electron microscope, or spectroscope have roughly divided the mechanism into three stages: (1) initial attachment; (2) microcolonies; and (3) maturation. However, these analytical methods are incapable to observe real-time changes in different locations of the extracellular matrix, and to analyze mechanical properties for single bacteria in micro and nanoscale. Since atomic force microscopy (AFM) operates by precise control of tip-sample interaction forces in liquid and in air, living microorganisms can be analyzed under near-physiological conditions. Thus, analytical techniques based on AFM constitute powerful tools for the study of biological samples, both qualitatively and quantitatively. In this study, we used AFM to quantitatively track the changes of multiple nanomechanical properties of S. mutans, including dissipation energy, adhesion force, deformation, and elastic modulus at different metabolic stages. The data revealed that the bacterial extracellular matrix has a gradient distribution in stickiness, in which different stickiness indicates the variation of EPS compositions, freshness, and metabolic stages. In-situ, time-lapse AFM images showed the local generation and distribution of EPS at different times, in which the highest adhesion distributed along sides of the S. mutans cells. Through time-lapse

  3. Spatiotemporal monitoring of soil water content profiles in an irrigated field using probabilistic inversion of time-lapse EMI data

    KAUST Repository

    Moghadas, Davood

    2017-10-17

    Monitoring spatiotemporal variations of soil water content (θ) is important across a range of research fields, including agricultural engineering, hydrology, meteorology and climatology. Low frequency electromagnetic induction (EMI) systems have proven to be useful tools in mapping soil apparent electrical conductivity (σa) and soil moisture. However, obtaining depth profile water content is an area that has not been fully explored using EMI. To examine this, we performed time-lapse EMI measurements using a CMD mini-Explorer sensor along a 10 m transect of a maize field over a 6 day period. Reference data were measured at the end of the profile via an excavated pit using 5TE capacitance sensors. In order to derive a time-lapse, depth-specific subsurface image of electrical conductivity (σ), we applied a probabilistic sampling approach, DREAM(ZS), on the measured EMI data. The inversely estimated σ values were subsequently converted to θ using the Rhoades et al. (1976) petrophysical relationship. The uncertainties in measured σa, as well as inaccuracies in the inverted data, introduced some discrepancies between estimated σ and reference values in time and space. Moreover, the disparity between the measurement footprints of the 5TE and CMD Mini-Explorer sensors also led to differences. The obtained θ permitted an accurate monitoring of the spatiotemporal distribution and variation of soil water content due to root water uptake and evaporation. The proposed EMI measurement and modeling technique also allowed for detecting temporal root zone soil moisture variations. The time-lapse θ monitoring approach developed using DREAM(ZS) thus appears to be a useful technique to understand spatiotemporal patterns of soil water content and provide insights into linked soil moisture vegetation processes and the dynamics of soil moisture/infiltration processes.

  4. Spatiotemporal monitoring of soil water content profiles in an irrigated field using probabilistic inversion of time-lapse EMI data

    Science.gov (United States)

    Moghadas, Davood; Jadoon, Khan Zaib; McCabe, Matthew F.

    2017-12-01

    Monitoring spatiotemporal variations of soil water content (θ) is important across a range of research fields, including agricultural engineering, hydrology, meteorology and climatology. Low frequency electromagnetic induction (EMI) systems have proven to be useful tools in mapping soil apparent electrical conductivity (σa) and soil moisture. However, obtaining depth profile water content is an area that has not been fully explored using EMI. To examine this, we performed time-lapse EMI measurements using a CMD mini-Explorer sensor along a 10 m transect of a maize field over a 6 day period. Reference data were measured at the end of the profile via an excavated pit using 5TE capacitance sensors. In order to derive a time-lapse, depth-specific subsurface image of electrical conductivity (σ), we applied a probabilistic sampling approach, DREAM(ZS) , on the measured EMI data. The inversely estimated σ values were subsequently converted to θ using the Rhoades et al. (1976) petrophysical relationship. The uncertainties in measured σa, as well as inaccuracies in the inverted data, introduced some discrepancies between estimated σ and reference values in time and space. Moreover, the disparity between the measurement footprints of the 5TE and CMD Mini-Explorer sensors also led to differences. The obtained θ permitted an accurate monitoring of the spatiotemporal distribution and variation of soil water content due to root water uptake and evaporation. The proposed EMI measurement and modeling technique also allowed for detecting temporal root zone soil moisture variations. The time-lapse θ monitoring approach developed using DREAM(ZS) thus appears to be a useful technique to understand spatiotemporal patterns of soil water content and provide insights into linked soil moisture vegetation processes and the dynamics of soil moisture/infiltration processes.

  5. Spatiotemporal monitoring of soil water content profiles in an irrigated field using probabilistic inversion of time-lapse EMI data

    KAUST Repository

    Moghadas, Davood; Jadoon, Khan Zaib; McCabe, Matthew

    2017-01-01

    Monitoring spatiotemporal variations of soil water content (θ) is important across a range of research fields, including agricultural engineering, hydrology, meteorology and climatology. Low frequency electromagnetic induction (EMI) systems have proven to be useful tools in mapping soil apparent electrical conductivity (σa) and soil moisture. However, obtaining depth profile water content is an area that has not been fully explored using EMI. To examine this, we performed time-lapse EMI measurements using a CMD mini-Explorer sensor along a 10 m transect of a maize field over a 6 day period. Reference data were measured at the end of the profile via an excavated pit using 5TE capacitance sensors. In order to derive a time-lapse, depth-specific subsurface image of electrical conductivity (σ), we applied a probabilistic sampling approach, DREAM(ZS), on the measured EMI data. The inversely estimated σ values were subsequently converted to θ using the Rhoades et al. (1976) petrophysical relationship. The uncertainties in measured σa, as well as inaccuracies in the inverted data, introduced some discrepancies between estimated σ and reference values in time and space. Moreover, the disparity between the measurement footprints of the 5TE and CMD Mini-Explorer sensors also led to differences. The obtained θ permitted an accurate monitoring of the spatiotemporal distribution and variation of soil water content due to root water uptake and evaporation. The proposed EMI measurement and modeling technique also allowed for detecting temporal root zone soil moisture variations. The time-lapse θ monitoring approach developed using DREAM(ZS) thus appears to be a useful technique to understand spatiotemporal patterns of soil water content and provide insights into linked soil moisture vegetation processes and the dynamics of soil moisture/infiltration processes.

  6. Observation of human embryonic behavior in vitro by high-resolution time-lapse cinematography.

    Science.gov (United States)

    Iwata, Kyoko; Mio, Yasuyuki

    2016-07-01

    Assisted reproductive technology (ART) has yielded vast amounts of information and knowledge on human embryonic development in vitro; however, still images provide limited data on dynamic changes in the developing embryos. Using our high-resolution time-lapse cinematography (hR-TLC) system, we were able to describe normal human embryonic development continuously from the fertilization process to the hatched blastocyst stage in detail. Our hR-TLC observation also showed the embryonic abnormality of a third polar body (PB)-like substance likely containing a small pronucleus being extruded and resulting in single-pronucleus (1PN) formation, while our molecular biological investigations suggested the possibility that some 1PN embryos could be diploid, carrying both maternal and paternal genomes. Furthermore, in some embryos the extruded third PB-like substance was eventually re-absorbed into the ooplasm resulting in the formation of an uneven-sized, two-PN zygote. In addition, other hR-TLC observations showed that cytokinetic failure was correlated with equal-sized, multi-nucleated blastomeres that were also observed in the embryo showing early initiation of compaction. Assessment combining our hR-TLC with molecular biological techniques enables a better understanding of embryonic development and potential improvements in ART outcomes.

  7. Reservoir characterization using production data and time-lapse seismic data

    Energy Technology Data Exchange (ETDEWEB)

    Dadashpour, Mohsen

    2009-12-15

    The most commonly encountered, and probably the most challenging task in reservoir engineering, is to describe the reservoir accurately and efficiently. An accurate description of a reservoir is crucial to the management of production and efficiency of oil recovery. Reservoir modeling is an important step in a reservoir's future performance, which is in direct proportion to reservoir management, risk analysis and making key economic decisions. The purpose of reservoir modeling is to not only build a model that is consistent with currently available data, but to build one that gives a good prediction of its future behavior. Updating a reservoir model to behave as closely as possible to the real reservoir is called history matching, and the estimation of reservoir properties using this method is known as parameter estimation problem, which is an inversion process. Parameter estimation is a time consuming and non-unique problem with a large solution space. Saturation and pressure changes, and porosity and permeability distributions are the most common parameters to estimate in the oil industry. These parameters must be specified in every node within a petroleum reservoir simulator. These parameters will be adjusted until the model prediction data match the observation data to a sufficient degree. The solution space reduction in this project is done by adding time-lapse seismic data as a new set of dynamic data to the traditional production histories. Time-lapse (or 4D) seismic consists of two or more 3D seismic surveys shot at different calendar times. Time-lapse seismic surveys produce images at different times in a reservoir's history. The seismic response of a reservoir may change due to changes in pressure, fluid saturation and temperature. These changes in seismic images due to a variation in saturation and pressure can be used as additional observation data. Time-lapse seismic data are dynamical measurements which have a high resolution in the

  8. Coherence revivals in two-photon frequency combs

    International Nuclear Information System (INIS)

    Torres-Company, Victor; Lancis, Jesus; Lajunen, Hanna; Friberg, Ari T.

    2011-01-01

    We describe and theoretically analyze the self-imaging Talbot effect of entangled photon pairs in the time domain. Rich phenomena are observed in coherence propagation along dispersive media of mode-locked two-photon states with frequency entanglement exhibiting a comblike correlation function. Our results can be used to remotely transfer frequency standards through optical fiber networks with two-photon light, avoiding the requirement of dispersion compensation.

  9. How does blastomere removal affect embryonic development? : A time-lapse analysis

    DEFF Research Database (Denmark)

    Kirkegaard, Kirstine; Hindkjær, Johnny Juhl; Ingerslev, Hans Jakob

    of the 6-10 cell embryo. It has been argued that blastomere removal does not affect embryonic development, but few studies have focussed on safety of the procedure. Recently, time-lapse studies on mice have suggested that blastomere removal affects embryonic development. The present study was conducted...... to evaluate the effect of blastomere biopsy on early human embryonic development using time-lapse analysis. Materials and methods: Couples undergoing IVF treatment or PGD were requested permission to include embryos in the project. The diagnosis healthy/diseased was made by analysis of a single blastomere....... For PGD 56 human embryos were biopsied 68 hours after fertilisation, the majority at the eight cell stage. As controls 43 non-biopsied embryos at the 6-8 cell stage were selected. All embryos were cultured until 5 days after fertilisation in a time-lapse incubator (EmbryoScope™). Key events such as time...

  10. Uniform silica nanoparticles encapsulating two-photon absorbing fluorescent dye

    International Nuclear Information System (INIS)

    Wu Weibing; Liu Chang; Wang Mingliang; Huang Wei; Zhou Shengrui; Jiang Wei; Sun Yueming; Cui Yiping; Xu Chunxinag

    2009-01-01

    We have prepared uniform silica nanoparticles (NPs) doped with a two-photon absorbing zwitterionic hemicyanine dye by reverse microemulsion method. Obvious solvatochromism on the absorption spectra of dye-doped NPs indicates that solvents can partly penetrate into the silica matrix and then affect the ground and excited state of dye molecules. For dye-doped NP suspensions, both one-photon and two-photon excited fluorescence are much stronger and recorded at shorter wavelength compared to those of free dye solutions with comparative overall dye concentration. This behavior is possibly attributed to the restricted twisted intramolecular charge transfer (TICT), which reduces fluorescence quenching when dye molecules are trapped in the silica matrix. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells with low cytotoxicity. - Graphical abstract: Water-soluble silica NPs doped with a two-photon absorbing zwitterionic hemicyanine dye were prepared. They were found of enhanced one-photon and two-photon excited fluorescence compared to free dye solutions. Images from two-photon laser scanning fluorescence microscopy demonstrate that the dye-doped silica NPs can be actively uptaken by Hela cells.

  11. Automated Ground-based Time-lapse Camera Monitoring of West Greenland ice sheet outlet Glaciers: Challenges and Solutions

    Science.gov (United States)

    Ahn, Y.; Box, J. E.; Balog, J.; Lewinter, A.

    2008-12-01

    Monitoring Greenland outlet glaciers using remotely sensed data has drawn a great attention in earth science communities for decades and time series analysis of sensory data has provided important variability information of glacier flow by detecting speed and thickness changes, tracking features and acquiring model input. Thanks to advancements of commercial digital camera technology and increased solid state storage, we activated automatic ground-based time-lapse camera stations with high spatial/temporal resolution in west Greenland outlet and collected one-hour interval data continuous for more than one year at some but not all sites. We believe that important information of ice dynamics are contained in these data and that terrestrial mono-/stereo-photogrammetry can provide theoretical/practical fundamentals in data processing along with digital image processing techniques. Time-lapse images over periods in west Greenland indicate various phenomenon. Problematic is rain, snow, fog, shadows, freezing of water on camera enclosure window, image over-exposure, camera motion, sensor platform drift, and fox chewing of instrument cables, and the pecking of plastic window by ravens. Other problems include: feature identification, camera orientation, image registration, feature matching in image pairs, and feature tracking. Another obstacle is that non-metric digital camera contains large distortion to be compensated for precise photogrammetric use. Further, a massive number of images need to be processed in a way that is sufficiently computationally efficient. We meet these challenges by 1) identifying problems in possible photogrammetric processes, 2) categorizing them based on feasibility, and 3) clarifying limitation and alternatives, while emphasizing displacement computation and analyzing regional/temporal variability. We experiment with mono and stereo photogrammetric techniques in the aide of automatic correlation matching for efficiently handling the enormous

  12. Time-Lapse and Slow-Motion Tracking of Temperature Changes: Response Time of a Thermometer

    Science.gov (United States)

    Moggio, L.; Onorato, P.; Gratton, L. M.; Oss, S.

    2017-01-01

    We propose the use of a smartphone based time-lapse and slow-motion video techniques together with tracking analysis as valuable tools for investigating thermal processes such as the response time of a thermometer. The two simple experimental activities presented here, suitable also for high school and undergraduate students, allow one to measure…

  13. Exploiting the airwave for time-lapse reservoir monitoring with CSEM on land

    NARCIS (Netherlands)

    Wirianto, M.; Mulder, W.A.; Slob, E.C.

    2011-01-01

    In the application of controlled source electromagnetics for reservoir monitoring on land, repeatability errors in the source will mask the time-lapse signal due to hydrocarbon production when recording surface data close to the source. We demonstrate that at larger distances, the airwave will still

  14. Time-Lapse Monitoring of Subsurface Fluid Flow using Parsimonious Seismic Interferometry

    KAUST Repository

    Hanafy, Sherif; Li, Jing; Schuster, Gerard T.

    2017-01-01

    of parsimonious seismic interferometry with the time-lapse mentoring idea with field examples, where we were able to record 30 different data sets within a 2-hour period. The recorded data are then processed to generate 30 snapshots that shows the spread of water

  15. Time-lapse misorientation maps for the analysis of electron backscatter diffraction data from evolving microstructures

    NARCIS (Netherlands)

    Wheeler, J.; Cross, A.; Drury, M.; Hough, R.M.; Mariani, E.; Piazolo, S.; Prior, D.J.

    2011-01-01

    A “time-lapse misorientation map” is defined here as a map which shows the orientation change at each point in an evolving crystalline microstructure between two different times. Electron backscatter diffraction data from in situ heating experiments can be used to produce such maps, which then

  16. Holographic Two-Photon Induced Photopolymerization

    Data.gov (United States)

    Federal Laboratory Consortium — Holographic two-photon-induced photopolymerization (HTPIP) offers distinct advantages over conventional one-photon-induced photopolymerization and current techniques...

  17. A randomized clinical trial comparing embryo culture in a conventional incubator with a time-lapse incubator

    DEFF Research Database (Denmark)

    Kirkegaard, Kirstine; Hindkjaer, Johnny Juhl; Grøndahl, Marie Louise

    2012-01-01

    Time-lapse monitoring allows for a flexible embryo evaluation and potentially provides new dynamic markers of embryo competence. Before introducing time-lapse monitoring in a clinical setting, the safety of the instrument must be properly documented. Accordingly, the aim of this study...

  18. Application of a linear finite-frequency theory to time-lapse crosswell tomography in ultrasonic and numerical experiments

    NARCIS (Netherlands)

    Spetzler, J.; Sijacic, D.; Wolf, K.H.A.A.

    2007-01-01

    Time-lapse seismic monitoring is the geophysical discipline whereby multiple data sets recorded at the same location but at different times are used to locate and quantify temporal changes in the elastic parameters of the subsurface. We validate a time-lapse monitoring method by crosswell tomography

  19. Advances and challenges in label-free nonlinear optical imaging using two-photon excitation fluorescence and second harmonic generation for cancer research

    NARCIS (Netherlands)

    Thomas, Giju; van Voskuilen, Johan; Gerritsen, Hans C.; Sterenborg, H. J. C. M.

    2014-01-01

    Nonlinear optical imaging (NLOI) has emerged to be a promising tool for bio-medical imaging in recent times. Among the various applications of NLOI, its utility is the most significant in the field of pre-clinical and clinical cancer research. This review begins by briefly covering the core

  20. Quarkonium two-photon decays in QCD

    International Nuclear Information System (INIS)

    Dulyan, L.S.; Khodjamirian, A.Yu.; Magakian, A.D.

    1989-01-01

    The two-photon decay of tensor charmonium χ c2 → 2 γ is calculated with account of gluon condensate effects. The result is in good agreement with experiment. The two-photon width of pseudoscalar b-quarkonium η b → 2 γ is estimated. 19 refs.; 1 fig.; 1 tab

  1. A Novel Methodology for Characterizing Cell Subpopulations in Automated Time-lapse Microscopy

    Directory of Open Access Journals (Sweden)

    Georges Hattab

    2018-02-01

    Full Text Available Time-lapse imaging of cell colonies in microfluidic chambers provides time series of bioimages, i.e., biomovies. They show the behavior of cells over time under controlled conditions. One of the main remaining bottlenecks in this area of research is the analysis of experimental data and the extraction of cell growth characteristics, such as lineage information. The extraction of the cell line by human observers is time-consuming and error-prone. Previously proposed methods often fail because of their reliance on the accurate detection of a single cell, which is not possible for high density, high diversity of cell shapes and numbers, and high-resolution images with high noise. Our task is to characterize subpopulations in biomovies. In order to shift the analysis of the data from individual cell level to cellular groups with similar fluorescence or even subpopulations, we propose to represent the cells by two new abstractions: the particle and the patch. We use a three-step framework: preprocessing, particle tracking, and construction of the patch lineage. First, preprocessing improves the signal-to-noise ratio and spatially aligns the biomovie frames. Second, cell sampling is performed by assuming particles, which represent a part of a cell, cell or group of contiguous cells in space. Particle analysis includes the following: particle tracking, trajectory linking, filtering, and color information, respectively. Particle tracking consists of following the spatiotemporal position of a particle and gives rise to coherent particle trajectories over time. Typical tracking problems may occur (e.g., appearance or disappearance of cells, spurious artifacts. They are effectively processed using trajectory linking and filtering. Third, the construction of the patch lineage consists in joining particle trajectories that share common attributes (i.e., proximity and fluorescence intensity and feature common ancestry. This step is based on patch finding

  2. Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring

    DEFF Research Database (Denmark)

    Ingerslev, Hans Jakob; Hindkjær, Johnny Juhl; Kirkegaard, Kirstine

    2012-01-01

    -points for each cell division and blastocyst stages were registered until 120 hours after oocyte retrieval. Only 2PN embryos completing the first cleavage were evaluated. The groups were compared using one-way ANOVA or Kruskall-Wallis test. Estimates are reported as medians with 95% confidence intervals. Time......Introduction: Data from a number of studies indicate -but not unequivocally- that culture of embryos in 5% O2 compared to 20% O2 improves blastocyst formation in humans and various animal species and may yield better pregnancy rates in IVF. The detrimental effects of atmospheric oxygen were...... was to evaluate the influence of oxygen tension on human pre-implantation development using time-lapse monitoring. Materials and methods: Human embryos were cultured to the blastocyst stage in a time-lapse incubator (EmbryoScope™) in 20% O2 (group 1), 20% O2 for 24 hours followed by culture in 5% O2 (group 2...

  3. Time-Lapse Monitoring of Subsurface Fluid Flow using Parsimonious Seismic Interferometry

    KAUST Repository

    Hanafy, Sherif

    2017-04-21

    A typical small-scale seismic survey (such as 240 shot gathers) takes at least 16 working hours to be completed, which is a major obstacle in case of time-lapse monitoring experiments. This is especially true if the subject that needs to be monitored is rapidly changing. In this work, we will discuss how to decrease the recording time from 16 working hours to less than one hour of recording. Here, the virtual data has the same accuracy as the conventional data. We validate the efficacy of parsimonious seismic interferometry with the time-lapse mentoring idea with field examples, where we were able to record 30 different data sets within a 2-hour period. The recorded data are then processed to generate 30 snapshots that shows the spread of water from the ground surface down to a few meters.

  4. Capturing tissue repair in zebrafish larvae with time-lapse brightfield stereomicroscopy.

    Science.gov (United States)

    Lisse, Thomas S; Brochu, Elizabeth A; Rieger, Sandra

    2015-01-31

    The zebrafish larval tail fin is ideal for studying tissue regeneration due to the simple architecture of the larval fin-fold, which comprises of two layers of skin that enclose undifferentiated mesenchyme, and because the larval tail fin regenerates rapidly within 2-3 days. Using this system, we demonstrate a method for capturing the repair dynamics of the amputated tail fin with time-lapse video brightfield stereomicroscopy. We demonstrate that fin amputation triggers a contraction of the amputation wound and extrusion of cells around the wound margin, leading to their subsequent clearance. Fin regeneration proceeds from proximal to distal direction after a short delay. In addition, developmental growth of the larva can be observed during all stages. The presented method provides an opportunity for observing and analyzing whole tissue-scale behaviors such as fin development and growth in a simple microscope setting, which is easily adaptable to any stereomicroscope with time-lapse capabilities.

  5. Freeze core sampling to validate time-lapse resistivity monitoring of the hyporheic zone.

    Science.gov (United States)

    Toran, Laura; Hughes, Brian; Nyquist, Jonathan; Ryan, Robert

    2013-01-01

    A freeze core sampler was used to characterize hyporheic zone storage during a stream tracer test. The pore water from the frozen core showed tracer lingered in the hyporheic zone after the tracer had returned to background concentration in collocated well samples. These results confirmed evidence of lingering subsurface tracer seen in time-lapse electrical resistivity tomographs. The pore water exhibited brine exclusion (ion concentrations in ice lower than source water) in a sediment matrix, despite the fast freezing time. Although freeze core sampling provided qualitative evidence of lingering tracer, it proved difficult to quantify tracer concentration because the amount of brine exclusion during freezing could not be accurately determined. Nonetheless, the additional evidence for lingering tracer supports using time-lapse resistivity to detect regions of low fluid mobility within the hyporheic zone that can act as chemically reactive zones of importance in stream health. © 2012, The Author(s). GroundWater © 2012, National Ground Water Association.

  6. Studying Axon-Astrocyte Functional Interactions by 3D Two-Photon Ca2+ Imaging: A Practical Guide to Experiments and "Big Data" Analysis.

    Science.gov (United States)

    Savtchouk, Iaroslav; Carriero, Giovanni; Volterra, Andrea

    2018-01-01

    Recent advances in fast volumetric imaging have enabled rapid generation of large amounts of multi-dimensional functional data. While many computer frameworks exist for data storage and analysis of the multi-gigabyte Ca 2+ imaging experiments in neurons, they are less useful for analyzing Ca 2+ dynamics in astrocytes, where transients do not follow a predictable spatio-temporal distribution pattern. In this manuscript, we provide a detailed protocol and commentary for recording and analyzing three-dimensional (3D) Ca 2+ transients through time in GCaMP6f-expressing astrocytes of adult brain slices in response to axonal stimulation, using our recently developed tools to perform interactive exploration, filtering, and time-correlation analysis of the transients. In addition to the protocol, we release our in-house software tools and discuss parameters pertinent to conducting axonal stimulation/response experiments across various brain regions and conditions. Our software tools are available from the Volterra Lab webpage at https://wwwfbm.unil.ch/dnf/group/glia-an-active-synaptic-partner/member/volterra-andrea-volterra in the form of software plugins for Image J (NIH)-a de facto standard in scientific image analysis. Three programs are available: MultiROI_TZ_profiler for interactive graphing of several movable ROIs simultaneously, Gaussian_Filter5D for Gaussian filtering in several dimensions, and Correlation_Calculator for computing various cross-correlation parameters on voxel collections through time.

  7. Informing hydrological models with ground-based time-lapse relative gravimetry: potential and limitations

    DEFF Research Database (Denmark)

    Bauer-Gottwein, Peter; Christiansen, Lars; Rosbjerg, Dan

    2011-01-01

    parameter uncertainty decreased significantly when TLRG data was included in the inversion. The forced infiltration experiment caused changes in unsaturated zone storage, which were monitored using TLRG and ground-penetrating radar. A numerical unsaturated zone model was subsequently conditioned on both......Coupled hydrogeophysical inversion emerges as an attractive option to improve the calibration and predictive capability of hydrological models. Recently, ground-based time-lapse relative gravity (TLRG) measurements have attracted increasing interest because there is a direct relationship between...

  8. Assessment of Time-Lapse in Visible and Thermal Face Recognition

    Czech Academy of Sciences Publication Activity Database

    Farokhi, Sajad; Shamsuddin, Siti Mariyam; Flusser, Jan; Sheikh, Usman Ullah

    2012-01-01

    Roč. 6, č. 1 (2012), s. 181-186 R&D Projects: GA ČR GAP103/11/1552 Institutional support: RVO:67985556 Keywords : face recognition * moment invariants * Zernike moments Subject RIV: JD - Computer Applications, Robotics http://library.utia.cas.cz/separaty/2012/ZOI/flusser-assessment of time-lapse in visible and thermal face recognition -j.pdf

  9. Unscented Kalman filter assimilation of time-lapse self-potential data for monitoring solute transport

    Science.gov (United States)

    Cui, Yi-an; Liu, Lanbo; Zhu, Xiaoxiong

    2017-08-01

    Monitoring the extent and evolution of contaminant plumes in local and regional groundwater systems from existing landfills is critical in contamination control and remediation. The self-potential survey is an efficient and economical nondestructive geophysical technique that can be used to investigate underground contaminant plumes. Based on the unscented transform, we have built a Kalman filtering cycle to conduct time-lapse data assimilation for monitoring the transport of solute based on the solute transport experiment using a bench-scale physical model. The data assimilation was formed by modeling the evolution based on the random walk model and observation correcting based on the self-potential forward. Thus, monitoring self-potential data can be inverted by the data assimilation technique. As a result, we can reconstruct the dynamic process of the contaminant plume instead of using traditional frame-to-frame static inversion, which may cause inversion artifacts. The data assimilation inversion algorithm was evaluated through noise-added synthetic time-lapse self-potential data. The result of the numerical experiment shows validity, accuracy and tolerance to the noise of the dynamic inversion. To validate the proposed algorithm, we conducted a scaled-down sandbox self-potential observation experiment to generate time-lapse data that closely mimics the real-world contaminant monitoring setup. The results of physical experiments support the idea that the data assimilation method is a potentially useful approach for characterizing the transport of contamination plumes using the unscented Kalman filter (UKF) data assimilation technique applied to field time-lapse self-potential data.

  10. Evaluating four-dimensional time-lapse electrical resistivity tomography for monitoring DNAPL source zone remediation.

    Science.gov (United States)

    Power, Christopher; Gerhard, Jason I; Karaoulis, Marios; Tsourlos, Panagiotis; Giannopoulos, Antonios

    2014-07-01

    Practical, non-invasive tools do not currently exist for mapping the remediation of dense non-aqueous phase liquids (DNAPLs). Electrical resistivity tomography (ERT) exhibits significant potential but has not yet become a practitioner's tool due to challenges in interpreting the survey results at real sites. This study explores the effectiveness of recently developed four-dimensional (4D, i.e., 3D space plus time) time-lapse surface ERT to monitor DNAPL source zone remediation. A laboratory experiment demonstrated the approach for mapping a changing NAPL distribution over time. A recently developed DNAPL-ERT numerical model was then employed to independently simulate the experiment, providing confidence that the DNAPL-ERT model is a reliable tool for simulating real systems. The numerical model was then used to evaluate the potential for this approach at the field scale. Four DNAPL source zones, exhibiting a range of complexity, were initially simulated, followed by modeled time-lapse ERT monitoring of complete DNAPL remediation by enhanced dissolution. 4D ERT inversion provided estimates of the regions of the source zone experiencing mass reduction with time. Results show that 4D time-lapse ERT has significant potential to map both the outline and the center of mass of the evolving treated portion of the source zone to within a few meters in each direction. In addition, the technique can provide a reasonable, albeit conservative, estimate of the DNAPL volume remediated with time: 25% underestimation in the upper 2m and up to 50% underestimation at late time between 2 and 4m depth. The technique is less reliable for identifying cleanup of DNAPL stringers outside the main DNAPL body. Overall, this study demonstrates that 4D time-lapse ERT has potential for mapping where and how quickly DNAPL mass changes in real time during site remediation. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Time-lapse electrical surveys to locate infiltration zones in weathered hard rock tropical areas

    Science.gov (United States)

    Wubda, M.; Descloitres, M.; Yalo, N.; Ribolzi, O.; Vouillamoz, J. M.; Boukari, M.; Hector, B.; Séguis, L.

    2017-07-01

    In West Africa, infiltration and groundwater recharge processes in hard rock areas are depending on climatic, surface and subsurface conditions, and are poorly documented. Part of the reason is that identification, location and monitoring of these processes is still a challenge. Here, we explore the potential for time-lapse electrical surveys to bring additional information on these processes for two different climate situations: a semi-arid Sahelian site (north of Burkina and a humid Sudanian site (north of Benin), respectively focusing on indirect (localized) and direct (diffuse) recharge processes. The methodology is based on surveys in dry season and rainy season on typical pond or gully using Electrical Resistivity Tomography (ERT) and frequency electromagnetic (FEM) apparent conductivity mapping. The results show that in the Sahelian zone an indirect recharge occurs as expected, but infiltration doesn't takes place at the center of the pond to the aquifer, but occurs laterally in the banks. In Sudanian zone, the ERT survey shows a direct recharge process as expected, but also a complicated behavior of groundwater dilution, as well as the role of hardpans for fast infiltration. These processes are ascertained by groundwater monitoring in adjacent observing wells. At last, FEM time lapse mapping is found to be difficult to quantitatively interpreted due to the non-uniqueness of the model, clearly evidenced comparing FEM result to auger holes monitoring. Finally, we found that time-lapse ERT can be an efficient way to track infiltration processes across ponds and gullies in both climatic conditions, the Sahelian setting providing results easier to interpret, due to significant resistivity contrasts between dry and rain seasons. Both methods can be used for efficient implementation of punctual sensors for complementary studies. However, FEM time-lapse mapping remains difficult to practice without external information that renders this method less attractive for

  12. Time-lapse seismic - repeatability versus usefulness and 2D versus 3D

    Science.gov (United States)

    Landro, M.

    2017-12-01

    Time-lapse seismic has developed rapidly over the past decades, especially for monitoring of oil and gas reservoirs and subsurface storage of CO2. I will review and discuss some of the critical enabling factors for the commercial success of this technology. It was early realized that how well we are able to repeat our seismic experiment is crucial. However, it is always a question of detectability versus repeatability. For marine seismic, there are several factors limiting the repeatability: Weather conditions, positioning of sources and receivers and so on. I will discuss recent improvements in both acquisition and processing methods over the last decade. It is well known that repeated 3D seismic data is the most accurate tool for reservoir monitoring purposes. However, several examples show that 2D seismic data may be used for monitoring purposes despite lower repeatability. I will use examples from an underground blow out in the North Sea, and repeated 2D seismic lines acquired before and after the Tohoku earthquake in 2011 to illustrate this. A major challenge when using repeated 2D seismic for subsurface monitoring purposes is the lack of 3D calibration points and significantly less amount of data. For marine seismic acquisition, feathering issues and crossline dip effects become more critical compared to 3D seismic acquisition. Furthermore, the uncertainties arising from a non-ideal 2D seismic acquisition are hard to assess, since the 3D subsurface geometry has not been mapped. One way to shed more light on this challenge is to use 3D time lapse seismic modeling testing various crossline dips or geometries. Other ways are to use alternative data sources, such as bathymetry, time lapse gravity or electromagnetic data. The end result for all time-lapse monitoring projects is an interpretation associated with uncertainties, and for the 2D case these uncertainties are often large. The purpose of this talk is to discuss how to reduces and control these

  13. Calixarenes and cations: a time-lapse photography of the big-bang.

    Science.gov (United States)

    Casnati, Alessandro

    2013-08-07

    The outstanding cation complexation properties emerging from the pioneering studies on calixarene ligands during a five-year period in the early 1980s triggered a big-bang burst of publications on such macrocycles that is still lasting at a distance of more than 30 years. A time-lapse photography of this timeframe is proposed which allows the readers to pinpoint the contributions of the different research groups.

  14. Time-lapse misorientation maps for the analysis of electron backscatter diffraction data from evolving microstructures

    International Nuclear Information System (INIS)

    Wheeler, J.; Cross, A.; Drury, M.; Hough, R.M.; Mariani, E.; Piazolo, S.; Prior, D.J.

    2011-01-01

    A 'time-lapse misorientation map' is defined here as a map which shows the orientation change at each point in an evolving crystalline microstructure between two different times. Electron backscatter diffraction data from in situ heating experiments can be used to produce such maps, which then highlight areas of microstructural change and also yield statistics indicative of how far different types of boundary (with different misorientations) have moved.

  15. Inversion of time-domain induced polarization data based on time-lapse concept

    Science.gov (United States)

    Kim, Bitnarae; Nam, Myung Jin; Kim, Hee Joon

    2018-05-01

    Induced polarization (IP) surveys, measuring overvoltage phenomena of the medium, are widely and increasingly performed not only for exploration of mineral resources but also for engineering applications. Among several IP survey methods such as time-domain, frequency-domain and spectral IP surveys, this study introduces a noble inversion method for time-domain IP data to recover the chargeability structure of target medium. The inversion method employs the concept of 4D inversion of time-lapse resistivity data sets, considering the fact that measured voltage in time-domain IP survey is distorted by IP effects to increase from the instantaneous voltage measured at the moment the source current injection starts. Even though the increase is saturated very fast, we can consider the saturated and instantaneous voltages as a time-lapse data set. The 4D inversion method is one of the most powerful method for inverting time-lapse resistivity data sets. Using the developed IP inversion algorithm, we invert not only synthetic but also field IP data to show the effectiveness of the proposed method by comparing the recovered chargeability models with those from linear inversion that was used for the inversion of the field data in a previous study. Numerical results confirm that the proposed inversion method generates reliable chargeability models even though the anomalous bodies have large IP effects.

  16. Scanless two-photon excitation of channelrhodopsin-2

    DEFF Research Database (Denmark)

    Papagiakoumou, E.; Anselmi, F.; Bègue, A.

    2010-01-01

    developed a method that combines generalized phase contrast with temporal focusing (TF-GPC) to shape two-photon excitation for this purpose. The illumination patterns are generated automatically from fluorescence images of neurons and shaped to cover the cell body or dendrites, or distributed groups...... of cells. The TF-GPC two-photon excitation patterns generated large photocurrents in Channelrhodopsin-2–expressing cultured cells and neurons and in mouse acute cortical slices. The amplitudes of the photocurrents can be precisely modulated by controlling the size and shape of the excitation volume and...

  17. Characterization for capillary barriers effects in a sand box test using time-lapsed GPR measurements

    Science.gov (United States)

    Kuroda, S.; Ishii, N.; Morii, T.

    2017-12-01

    Capillary barriers have been known as the method to protect subsurface regions against infiltration from soil surface. It is caused by essentially heterogeneous structure in permeability or soil physical property and produce non-uniform infiltration process then, in order to estimate the actual situation of the capillary barrier effect, the site-characterization with imaging technique like geophysical prospecting is effective. In this study, we examine the applicability of GPR to characterization for capillary barriers. We built a sand box with 90x340x90cm in which a thin high-permeable gravel layer was embedded as a capillary barrier. We conducted an infiltration test in the sand box using porous tube array for irrigation. It is expected to lead to non-uniform flow of soil water induced by capillary barrier effects. We monitored this process by various types of GPR measurements, including time-lapsed common offset profiling (COP) with multi- frequency antenna and transmission measurements like cross-borehole radar. At first, we conducted GPR common-offset survey. It could show the depth of capillary barrier in sand box. After that we conducted the infiltration test and GPR monitoring for infiltration process. GPR profiles can detect the wetting front and estimate water content change in the soil layer above the capillary barrier. From spatial change in these results we can estimate the effect of capillary barrier and the zone where the break through occur or not. Based on these results, we will discuss the applicability of GPR for monitoring the phenomena around the capillary barrier of soil. At first, we conducted GPR common-offset survey. It could show the depth of capillary barrier in sand box. After that we conducted the infiltration test and GPR monitoring for infiltration process. GPR profiles can detect the wetting front and estimate water content change in the soil layer above the capillary barrier. From spatial change in these results we can estimate the

  18. Inter-laboratory agreement on embryo classification and clinical decision: Conventional morphological assessment vs. time lapse.

    Science.gov (United States)

    Martínez-Granados, Luis; Serrano, María; González-Utor, Antonio; Ortíz, Nereyda; Badajoz, Vicente; Olaya, Enrique; Prados, Nicolás; Boada, Montse; Castilla, Jose A

    2017-01-01

    The aim of this study is to determine inter-laboratory variability on embryo assessment using time-lapse platform and conventional morphological assessment. This study compares the data obtained from a pilot study of external quality control (EQC) of time lapse, performed in 2014, with the classical EQC of the Spanish Society for the Study of Reproductive Biology (ASEBIR) performed in 2013 and 2014. In total, 24 laboratories (8 using EmbryoScope™, 15 using Primo Vision™ and one with both platforms) took part in the pilot study. The clinics that used EmbryoScope™ analysed 31 embryos and those using Primo Vision™ analysed 35. The classical EQC was implemented by 39 clinics, based on an analysis of 25 embryos per year. Both groups were required to evaluate various qualitative morphological variables (cell fragmentation, the presence of vacuoles, blastomere asymmetry and multinucleation), to classify the embryos in accordance with ASEBIR criteria and to stipulate the clinical decision taken. In the EQC time-lapse pilot study, the groups were asked to determine, as well as the above characteristics, the embryo development times, the number, opposition and size of pronuclei, the direct division of 1 into 3 cells and/or of 3 into 5 cells and false divisions. The degree of agreement was determined by calculating the intra-class correlation coefficients and the coefficient of variation for the quantitative variables and the Gwet index for the qualitative variables. For both EmbryoScope™ and Primo Vision™, two periods of greater inter-laboratory variability were observed in the times of embryo development events. One peak of variability was recorded among the laboratories addressing the first embryo events (extrusion of the second polar body and the appearance of pronuclei); the second peak took place between the times corresponding to the 8-cell and morula stages. In most of the qualitative variables analysed regarding embryo development, there was almost

  19. Inter-laboratory agreement on embryo classification and clinical decision: Conventional morphological assessment vs. time lapse.

    Directory of Open Access Journals (Sweden)

    Luis Martínez-Granados

    Full Text Available The aim of this study is to determine inter-laboratory variability on embryo assessment using time-lapse platform and conventional morphological assessment. This study compares the data obtained from a pilot study of external quality control (EQC of time lapse, performed in 2014, with the classical EQC of the Spanish Society for the Study of Reproductive Biology (ASEBIR performed in 2013 and 2014. In total, 24 laboratories (8 using EmbryoScope™, 15 using Primo Vision™ and one with both platforms took part in the pilot study. The clinics that used EmbryoScope™ analysed 31 embryos and those using Primo Vision™ analysed 35. The classical EQC was implemented by 39 clinics, based on an analysis of 25 embryos per year. Both groups were required to evaluate various qualitative morphological variables (cell fragmentation, the presence of vacuoles, blastomere asymmetry and multinucleation, to classify the embryos in accordance with ASEBIR criteria and to stipulate the clinical decision taken. In the EQC time-lapse pilot study, the groups were asked to determine, as well as the above characteristics, the embryo development times, the number, opposition and size of pronuclei, the direct division of 1 into 3 cells and/or of 3 into 5 cells and false divisions. The degree of agreement was determined by calculating the intra-class correlation coefficients and the coefficient of variation for the quantitative variables and the Gwet index for the qualitative variables. For both EmbryoScope™ and Primo Vision™, two periods of greater inter-laboratory variability were observed in the times of embryo development events. One peak of variability was recorded among the laboratories addressing the first embryo events (extrusion of the second polar body and the appearance of pronuclei; the second peak took place between the times corresponding to the 8-cell and morula stages. In most of the qualitative variables analysed regarding embryo development, there

  20. Charm production in two-photon collisions

    International Nuclear Information System (INIS)

    Linde, F.L.

    1988-01-01

    The analysis focuses on the production of charmonium resonances η c , χ 0 and χ 2 in two-photon interactions. The measurement of the inclusive production of charged D* mesons is described. 97 refs.; 54 figs.; 15 tabs

  1. Exclusive hadron production in two photon reactions

    International Nuclear Information System (INIS)

    Poppe, M.

    1986-02-01

    This paper summarises experimental results on exclusive hadron production in two photon collisions at electron positron storage rings and attempts some interpretation. Experimental know how is described and new suggestions are made for future analyses. New model calculations on resonance form factors and pair production amplitudes are presented. The two photon vertex is decomposed such that experiments can be parameterised with the minimal number of free parameters. Selection rules for off shell photon collisions are given in addition to Yang's theorems. (orig.)

  2. Stimulated emission depletion following two photon excitation

    OpenAIRE

    Marsh, R. J.; Armoogum, D. A.; Bain, A. J.

    2002-01-01

    The technique of stimulated emission depletion of fluorescence (STED) from a two photon excited molecular population is demonstrated in the S, excited state of fluorescein in ethylene glycol and methanol. Two photon excitation (pump) is achieved using the partial output of a regeneratively amplified Ti:Sapphire laser in conjunction with an optical parametric amplifier whose tuneable output provides a synchronous depletion (dump) pulse. Time resolved fluorescence intensity and anisotropy measu...

  3. Technical note: Stage and water width measurement of a mountain stream using a simple time-lapse camera

    Directory of Open Access Journals (Sweden)

    P. Leduc

    2018-01-01

    Full Text Available Remote sensing applied to river monitoring adds complementary information useful for understanding the system behaviour. In this paper, we present a method for visual stage gauging and water surface width measurement using a ground-based time-lapse camera and a fully automatic image analysis algorithm for flow monitoring at a river cross section of a steep, bouldery channel. The remote stage measurement was coupled with a water level logger (pressure transducer on site and shows that the image-based method gives a reliable estimate of the water height variation and daily flow record when validated against the pressure transducer (R = 0.91. From the remotely sensed pictures, we also extracted the water width and show that it is possible to correlate water surface width and stage. The images also provide valuable ancillary information for interpreting and understanding flow hydraulics and site weather conditions. This image-based gauging method is a reliable, informative and inexpensive alternative or adjunct to conventional stage measurement especially for remote sites.

  4. Time-lapse refraction seismic tomography for the detection of ground ice degradation

    Directory of Open Access Journals (Sweden)

    C. Hilbich

    2010-07-01

    Full Text Available The ice content of the subsurface is a major factor controlling the natural hazard potential of permafrost degradation in alpine terrain. Monitoring of changes in ice content is therefore similarly important as temperature monitoring in mountain permafrost. Although electrical resistivity tomography monitoring (ERTM proved to be a valuable tool for the observation of ice degradation, results are often ambiguous or contaminated by inversion artefacts. In theory, the sensitivity of P-wave velocity of seismic waves to phase changes between unfrozen water and ice is similar to the sensitivity of electric resistivity. Provided that the general conditions (lithology, stratigraphy, state of weathering, pore space remain unchanged over the observation period, temporal changes in the observed travel times of repeated seismic measurements should indicate changes in the ice and water content within the pores and fractures of the subsurface material. In this paper, a time-lapse refraction seismic tomography (TLST approach is applied as an independent method to ERTM at two test sites in the Swiss Alps. The approach was tested and validated based on a the comparison of time-lapse seismograms and analysis of reproducibility of the seismic signal, b the analysis of time-lapse travel time curves with respect to shifts in travel times and changes in P-wave velocities, and c the comparison of inverted tomograms including the quantification of velocity changes. Results show a high potential of the TLST approach concerning the detection of altered subsurface conditions caused by freezing and thawing processes. For velocity changes on the order of 3000 m/s even an unambiguous identification of significant ice loss is possible.

  5. Time-Lapse Electrical Geophysical Monitoring of Amendment-Based Biostimulation.

    Science.gov (United States)

    Johnson, Timothy C; Versteeg, Roelof J; Day-Lewis, Frederick D; Major, William; Lane, John W

    2015-01-01

    Biostimulation is increasingly used to accelerate microbial remediation of recalcitrant groundwater contaminants. Effective application of biostimulation requires successful emplacement of amendment in the contaminant target zone. Verification of remediation performance requires postemplacement assessment and contaminant monitoring. Sampling-based approaches are expensive and provide low-density spatial and temporal information. Time-lapse electrical resistivity tomography (ERT) is an effective geophysical method for determining temporal changes in subsurface electrical conductivity. Because remedial amendments and biostimulation-related biogeochemical processes often change subsurface electrical conductivity, ERT can complement and enhance sampling-based approaches for assessing emplacement and monitoring biostimulation-based remediation. Field studies demonstrating the ability of time-lapse ERT to monitor amendment emplacement and behavior were performed during a biostimulation remediation effort conducted at the Department of Defense Reutilization and Marketing Office (DRMO) Yard, in Brandywine, Maryland, United States. Geochemical fluid sampling was used to calibrate a petrophysical relation in order to predict groundwater indicators of amendment distribution. The petrophysical relations were field validated by comparing predictions to sequestered fluid sample results, thus demonstrating the potential of electrical geophysics for quantitative assessment of amendment-related geochemical properties. Crosshole radar zero-offset profile and borehole geophysical logging were also performed to augment the data set and validate interpretation. In addition to delineating amendment transport in the first 10 months after emplacement, the time-lapse ERT results show later changes in bulk electrical properties interpreted as mineral precipitation. Results support the use of more cost-effective surface-based ERT in conjunction with limited field sampling to improve spatial

  6. A state-space Bayesian framework for estimating biogeochemical transformations using time-lapse geophysical data

    Energy Technology Data Exchange (ETDEWEB)

    Chen, J.; Hubbard, S.; Williams, K.; Pride, S.; Li, L.; Steefel, C.; Slater, L.

    2009-04-15

    We develop a state-space Bayesian framework to combine time-lapse geophysical data with other types of information for quantitative estimation of biogeochemical parameters during bioremediation. We consider characteristics of end-products of biogeochemical transformations as state vectors, which evolve under constraints of local environments through evolution equations, and consider time-lapse geophysical data as available observations, which could be linked to the state vectors through petrophysical models. We estimate the state vectors and their associated unknown parameters over time using Markov chain Monte Carlo sampling methods. To demonstrate the use of the state-space approach, we apply it to complex resistivity data collected during laboratory column biostimulation experiments that were poised to precipitate iron and zinc sulfides during sulfate reduction. We develop a petrophysical model based on sphere-shaped cells to link the sulfide precipitate properties to the time-lapse geophysical attributes and estimate volume fraction of the sulfide precipitates, fraction of the dispersed, sulfide-encrusted cells, mean radius of the aggregated clusters, and permeability over the course of the experiments. Results of the case study suggest that the developed state-space approach permits the use of geophysical datasets for providing quantitative estimates of end-product characteristics and hydrological feedbacks associated with biogeochemical transformations. Although tested here on laboratory column experiment datasets, the developed framework provides the foundation needed for quantitative field-scale estimation of biogeochemical parameters over space and time using direct, but often sparse wellbore data with indirect, but more spatially extensive geophysical datasets.

  7. Time-lapse electrical geophysical monitoring of amendment-based biostimulation

    Science.gov (United States)

    Johnson, Timothy C.; Versteeg, Roelof J.; Day-Lewis, Frederick D.; Major, William; Lane, John W.

    2015-01-01

    Biostimulation is increasingly used to accelerate microbial remediation of recalcitrant groundwater contaminants. Effective application of biostimulation requires successful emplacement of amendment in the contaminant target zone. Verification of remediation performance requires postemplacement assessment and contaminant monitoring. Sampling-based approaches are expensive and provide low-density spatial and temporal information. Time-lapse electrical resistivity tomography (ERT) is an effective geophysical method for determining temporal changes in subsurface electrical conductivity. Because remedial amendments and biostimulation-related biogeochemical processes often change subsurface electrical conductivity, ERT can complement and enhance sampling-based approaches for assessing emplacement and monitoring biostimulation-based remediation.Field studies demonstrating the ability of time-lapse ERT to monitor amendment emplacement and behavior were performed during a biostimulation remediation effort conducted at the Department of Defense Reutilization and Marketing Office (DRMO) Yard, in Brandywine, Maryland, United States. Geochemical fluid sampling was used to calibrate a petrophysical relation in order to predict groundwater indicators of amendment distribution. The petrophysical relations were field validated by comparing predictions to sequestered fluid sample results, thus demonstrating the potential of electrical geophysics for quantitative assessment of amendment-related geochemical properties. Crosshole radar zero-offset profile and borehole geophysical logging were also performed to augment the data set and validate interpretation.In addition to delineating amendment transport in the first 10 months after emplacement, the time-lapse ERT results show later changes in bulk electrical properties interpreted as mineral precipitation. Results support the use of more cost-effective surface-based ERT in conjunction with limited field sampling to improve spatial

  8. Soil Contamination, Advanced integrated characterisation and time-lapse Monitoring, SoilCAM project highlights

    Science.gov (United States)

    French, H. K.; Van Der Zee, S. E.; Wehrer, M.; Godio, A.; Pedersen, L. B.; Tsocano, G.

    2013-12-01

    The SoilCAM project (2008- 2012, EU-FP7-212663) aimed at improving methods for monitoring subsurace contaminant distribution and biodegradation. Two test sites were chosen, Oslo airport Gardermoen, Norway where de-icing agents infiltrate the soil during snowmelt and the Trecate site in Italy where an inland crude oil spill occurred in 1994. A number of geophysical investigation techniques were combined with soil and water sampling techniques. Data obtained from time-lapse measurements were further analysed by numerical modelling of flow and transport at different scales in order to characterise transport processes in the unsaturated and saturated zones. Laboratory experiments provided physical and biogeochemical data for model parameterisation and to select remediation methods. The geophysical techniques were used to map geological heterogeneities and to conduct time-lapse measurements of processes in the unsaturated zone. Both cross borehole and surface electrodes were used for electrical resistivity and induced polarisation surveys. Results showed clear indications of areas highly affected by de-icing chemicals along the runway at Oslo airport. The time lapse measurements along the runway at the airport showed infiltration patterns during snowmelt and were used to validate 2D unsaturated flow and transport simulations using SUTRA. The simulations illustrate the effect of layering geological structures and membranes, buried parallel to the runway, on the flow pattern. Complex interaction between bio-geo-chemical processes in a 1D vertical profile along the runway were described with the ORCHESTRA model. Smaller scale field site measurements revealed increase of iron and manganese during degradation of de-icing chemicals. At the Trecate site a combination of georadar, electrical resistivity and radio magneto telluric provided a broad outline of the geology down to 50 m. Anomalies in the Induced polarisation and electrical resistivity data from the cross borehole

  9. Time-lapse reveals that osteoclasts can move across the bone surface while resorbing

    DEFF Research Database (Denmark)

    Søe, Kent; Delaissé, Jean-Marie

    2017-01-01

    , clear real-time observations are still lacking. Herein, we used specific markers and time-lapse to monitor live the spatiotemporal generation of resorption events by osteoclasts cultured on bone slices. In accordance with the current view, we found alternating episodes of resorption and migration...... trenches. Compared to pit events, trench events show properties enabling higher aggressiveness: long duration (days), high erosion speed (two times faster) and long-distance erosion (several 100 µm). Simultaneous resorption and migration reflect a unique situation where epithelial/secretory and mesenchymal....../migratory characteristics are integrated into just one cell phenotype, and deserves attention in future research....

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

    Science.gov (United States)

    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. Electrically Induced Two-Photon Transparency in Semiconductor Quantum Wells

    International Nuclear Information System (INIS)

    Hayat, Alex; Nevet, Amir; Orenstein, Meir

    2009-01-01

    We demonstrate experimentally two-photon transparency, achieved by current injection into a semiconductor quantum-well structure which exhibits two-photon emission. The two-photon induced luminescence is progressively reduced by the injected current, reaching the point of two-photon transparency - a necessary condition for semiconductor two-photon gain and lasing. These results agree with our calculations.

  12. Analysis of embryo morphokinetics, multinucleation and cleavage anomalies using continuous time-lapse monitoring in blastocyst transfer cycles.

    Science.gov (United States)

    Desai, Nina; Ploskonka, Stephanie; Goodman, Linnea R; Austin, Cynthia; Goldberg, Jeffrey; Falcone, Tommaso

    2014-06-20

    Time-lapse imaging combined with embryo morphokinetics may offer a non-invasive means for improving embryo selection. Data from clinics worldwide are necessary to compare and ultimately develop embryo classifications models using kinetic data. The primary objective of this study was to determine if there were kinetic differences between embryos with limited potential and those more often associated with in vitro blastocyst formation and/or implantation. We also wanted to compare putative kinetic markers for embryo selection as proposed by other laboratories to what we were observing in our own laboratory setting. Kinetic data and cycle outcomes were retrospectively analyzed in patients age 39 and younger with 7 or more zygotes cultured in the Embryoscope. Timing of specific events from the point of insemination were determined using time-lapse (TL) imaging. The following kinetic markers were assessed: time to syngamy (tPNf), t2, time to two cells (c), 3c (t3), 4c ( t4), 5c (t5), 8c (t8), morula (tMor), start of blastulation (tSB); tBL, blastocyst (tBL); expanded blastocyst (tEBL). Durations of the second (cc2) and third (cc3) cell cycles, the t5-t2 interval as well as time to complete synchronous divisions s1, s2 and s3 were calculated. Incidence and impact on development of nuclear and cleavage anomalies were also assessed. A total of 648 embryos transferred on day 5 were analyzed. The clinical pregnancy and implantation rate were 72% and 50%, respectively. Morphokinetic data showed that tPNf, t2,t4, t8, s1, s2,s3 and cc2 were significantly different in embryos forming blastocysts (ET or frozen) versus those with limited potential either failing to blastulate or else forming poor quality blastocysts ,ultimately discarded. Comparison of embryo kinetics in cycles with all embryos implanting (KID+) versus no implantation (KID-) suggested that markers of embryo competence to implant may be different from ability to form a blastocyst. The incidence of multinucleation

  13. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

    Science.gov (United States)

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-25

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  14. Time-lapse nanoscopy of friction in the non-Amontons and non-Coulomb regime.

    Science.gov (United States)

    Ishida, Tadashi; Sato, Takaaki; Ishikawa, Takahiro; Oguma, Masatsugu; Itamura, Noriaki; Goda, Keisuke; Sasaki, Naruo; Fujita, Hiroyuki

    2015-03-11

    Originally discovered by Leonard da Vinci in the 15th century, the force of friction is directly proportional to the applied load (known as Amontons' first law of friction). Furthermore, kinetic friction is independent of the sliding speed (known as Coulomb's law of friction). These empirical laws break down at high normal pressure (due to plastic deformation) and low sliding speed (in the transition regime between static friction and kinetic friction). An important example of this phenomenon is friction between the asperities of tectonic plates on the Earth. Despite its significance, little is known about the detailed mechanism of friction in this regime due to the lack of experimental methods. Here we demonstrate in situ time-lapse nanoscopy of friction between asperities sliding at ultralow speed (∼0.01 nm/s) under high normal pressure (∼GPa). This is made possible by compressing and rubbing a pair of nanometer-scale crystalline silicon anvils with electrostatic microactuators and monitoring its dynamical evolution with a transmission electron microscope. Our analysis of the time-lapse movie indicates that superplastic behavior is induced by decrystallization, plastic deformation, and atomic diffusion at the asperity-asperity interface. The results hold great promise for a better understanding of quasi-static friction under high pressure for geoscience, materials science, and nanotechnology.

  15. Using a time lapse microgravity model for mapping seawater intrusion around Semarang

    Energy Technology Data Exchange (ETDEWEB)

    Supriyadi,, E-mail: supriyadi@mail.unnes.ac.id; Khumaedi [Physics Department, Semarang State University (UNNES), D7 Building 2nd Floor FMIPA Sekaran Gunungpati (Indonesia); Yusuf, M. [Badan Meteologi Klimatologi Goefisika (BMKG), Jl.Angkasa I No.2 Kemayoran Jakarta Pusat (Indonesia); Agung, W. [Physics Department, Diponegoro University (UNDIP), Jl. Prof. Soedharto, Tembalang, Semarang (Indonesia)

    2016-03-11

    A modeling of time-lapse microgravity anomaly due to sea water intrusion has been conducted. It used field data of aquifer cross section, aquifer thickness and lithology of research area. Those data were then processed using Grav3D and Surfer. Modeling results indicated that the intrusion of sea water resulting in a time-lapse microgravity anomalies of 0.12 to 0.18 mGal, at soil layer density of 0.15 g/cm{sup 3} to 0.3 g/cm{sup 3} and at depth of 30 to 100 m. These imply that the areas experiencing seawater intrusion were Tanjung Mas, SPBE Bandarharjo, Brass, Old Market Boom and Johar as the microgravity measured there were in the range of 0.12 to 0.18 mGal and the density contrast were at 0.15 g/cm{sup 3} to 0.28 g/cm{sup 3}. Areas that experienced fluid reduction were Puri Anjasmoro, Kenconowungu and Puspowarno with microgravity changes from -0.06 mGal to -0.18 mGal.

  16. Stochastic modeling of oligodendrocyte generation in cell culture: model validation with time-lapse data

    Directory of Open Access Journals (Sweden)

    Noble Mark

    2006-05-01

    Full Text Available Abstract Background The purpose of this paper is two-fold. The first objective is to validate the assumptions behind a stochastic model developed earlier by these authors to describe oligodendrocyte generation in cell culture. The second is to generate time-lapse data that may help biomathematicians to build stochastic models of cell proliferation and differentiation under other experimental scenarios. Results Using time-lapse video recording it is possible to follow the individual evolutions of different cells within each clone. This experimental technique is very laborious and cannot replace model-based quantitative inference from clonal data. However, it is unrivalled in validating the structure of a stochastic model intended to describe cell proliferation and differentiation at the clonal level. In this paper, such data are reported and analyzed for oligodendrocyte precursor cells cultured in vitro. Conclusion The results strongly support the validity of the most basic assumptions underpinning the previously proposed model of oligodendrocyte development in cell culture. However, there are some discrepancies; the most important is that the contribution of progenitor cell death to cell kinetics in this experimental system has been underestimated.

  17. Time-lapse crystallography snapshots of a double-strand break repair polymerase in action.

    Science.gov (United States)

    Jamsen, Joonas A; Beard, William A; Pedersen, Lars C; Shock, David D; Moon, Andrea F; Krahn, Juno M; Bebenek, Katarzyna; Kunkel, Thomas A; Wilson, Samuel H

    2017-08-15

    DNA polymerase (pol) μ is a DNA-dependent polymerase that incorporates nucleotides during gap-filling synthesis in the non-homologous end-joining pathway of double-strand break repair. Here we report time-lapse X-ray crystallography snapshots of catalytic events during gap-filling DNA synthesis by pol μ. Unique catalytic intermediates and active site conformational changes that underlie catalysis are uncovered, and a transient third (product) metal ion is observed in the product state. The product manganese coordinates phosphate oxygens of the inserted nucleotide and PP i . The product metal is not observed during DNA synthesis in the presence of magnesium. Kinetic analyses indicate that manganese increases the rate constant for deoxynucleoside 5'-triphosphate insertion compared to magnesium. The likely product stabilization role of the manganese product metal in pol μ is discussed. These observations provide insight on structural attributes of this X-family double-strand break repair polymerase that impact its biological function in genome maintenance.DNA polymerase (pol) μ functions in DNA double-strand break repair. Here the authors use time-lapse X-ray crystallography to capture the states of pol µ during the conversion from pre-catalytic to product complex and observe a third transiently bound metal ion in the product state.

  18. The time-lapse AVO difference inversion for changes in reservoir parameters

    Science.gov (United States)

    Longxiao, Zhi; Hanming, Gu; Yan, Li

    2016-12-01

    The result of conventional time-lapse seismic processing is the difference between the amplitude and the post-stack seismic data. Although stack processing can improve the signal-to-noise ratio (SNR) of seismic data, it also causes a considerable loss of important information about the amplitude changes and only gives the qualitative interpretation. To predict the changes in reservoir fluid more precisely and accurately, we also need the quantitative information of the reservoir. To achieve this aim, we develop the method of time-lapse AVO (amplitude versus offset) difference inversion. For the inversion of reservoir changes in elastic parameters, we apply the Gardner equation as the constraint and convert the three-parameter inversion of elastic parameter changes into a two-parameter inversion to make the inversion more stable. For the inversion of variations in the reservoir parameters, we infer the relation between the difference of the reflection coefficient and variations in the reservoir parameters, and then invert reservoir parameter changes directly. The results of the theoretical modeling computation and practical application show that our method can estimate the relative variations in reservoir density, P-wave and S-wave velocity, calculate reservoir changes in water saturation and effective pressure accurately, and then provide reference for the rational exploitation of the reservoir.

  19. Time lapse seismic observations and effects of reservoir compressibility at Teal South oil field

    Science.gov (United States)

    Islam, Nayyer

    One of the original ocean-bottom time-lapse seismic studies was performed at the Teal South oil field in the Gulf of Mexico during the late 1990's. This work reexamines some aspects of previous work using modern analysis techniques to provide improved quantitative interpretations. Using three-dimensional volume visualization of legacy data and the two phases of post-production time-lapse data, I provide additional insight into the fluid migration pathways and the pressure communication between different reservoirs, separated by faults. This work supports a conclusion from previous studies that production from one reservoir caused regional pressure decline that in turn resulted in liberation of gas from multiple surrounding unproduced reservoirs. I also provide an explanation for unusual time-lapse changes in amplitude-versus-offset (AVO) data related to the compaction of the producing reservoir which, in turn, changed an isotropic medium to an anisotropic medium. In the first part of this work, I examine regional changes in seismic response due to the production of oil and gas from one reservoir. The previous studies primarily used two post-production ocean-bottom surveys (Phase I and Phase II), and not the legacy streamer data, due to the unavailability of legacy prestack data and very different acquisition parameters. In order to incorporate the legacy data in the present study, all three post-stack data sets were cross-equalized and examined using instantaneous amplitude and energy volumes. This approach appears quite effective and helps to suppress changes unrelated to production while emphasizing those large-amplitude changes that are related to production in this noisy (by current standards) suite of data. I examine the multiple data sets first by using the instantaneous amplitude and energy attributes, and then also examine specific apparent time-lapse changes through direct comparisons of seismic traces. In so doing, I identify time-delays that, when

  20. Analysis of compaction initiation in human embryos by using time-lapse cinematography.

    Science.gov (United States)

    Iwata, Kyoko; Yumoto, Keitaro; Sugishima, Minako; Mizoguchi, Chizuru; Kai, Yoshiteru; Iba, Yumiko; Mio, Yasuyuki

    2014-04-01

    To analyze the initiation of compaction in human embryos in vitro by using time-lapse cinematography (TLC), with the goal of determining the precise timing of compaction and clarifying the morphological changes underlying the compaction process. One hundred and fifteen embryos donated by couples with no further need for embryo-transfer were used in this study. Donated embryos were thawed and processed, and then their morphological behavior during the initiation of compaction was dynamically observed via time-lapse cinematography (TLC) for 5 days. Although the initiation of compaction occurred throughout the period from the 4-cell to 16-cell stage, 99 (86.1 %) embryos initiated compaction at the 8-cell stage or later, with initiation at the 8-cell stage being most frequent (22.6 %). Of these 99 embryos, 49.5 % developed into good-quality blastocysts. In contrast, of the 16 (13.9 %) embryos that initiated compaction prior to the 8-cell stage, only 18.8 % developed into good-quality blastocysts. Embryos that initiated compaction before the 8-cell stage showed significantly higher numbers of multinucleated blastomeres, due to asynchronism in nuclear division at the third mitotic division resulting from cytokinetic failure. The initiation of compaction primarily occurs at the third mitotic division or later in human embryos. Embryos that initiate compaction before the 8-cell stage are usually associated with aberrant embryonic development (i.e., cytokinetic failure accompanied by karyokinesis).

  1. Time Lapse Electrical Resistivity to Connect Evapotranspiration and Groundwater Fluxes in the Critical Zone

    Science.gov (United States)

    Jarvis, S. K.; Harmon, R. E.; Barnard, H. R.; Randall, J.; Singha, K.

    2017-12-01

    The critical zone (CZ)—an open system extending from canopy top to the base of groundwater—is a highly dynamic and heterogeneous environment. In forested terrain, trees make up a large component of the CZ. This work aims to quantify the connection between vegetation and subsurface water storage at a hillslope scale within a forested watershed in the H.J. Andrews Experimental Forest, Oregon. To identify the mechanism(s) controlling the connection at the hillslope scale, we observe patterns in electrical conductivity using 2D-time lapse-DC resistivity. To compare inversions through time a representative error model was determined using L-curve criterion. Inverted data show high spatial variability in ground electrical conductivity and variation at both diel and seasonal timescales. These changes are most pronounced in areas corresponding to dense vegetation. The diel pattern in electrical conductivity is also observed in monitored sap flow sensors, water-level gauges, tensiometers, and sediment thermal probes. To quantify the temporal connection between these data over the course of the growing season a cross correlation analysis was conducted. Preliminary data show that over the course of the growing season transpiration becomes decoupled from both groundwater and soil moisture. Further decomposition of the inverted time lapse data will highlight spatial variability in electrical conductivity providing insight into the where, when, and how(s) of tree-modified subsurface storage.

  2. Two-photon superradiance in extended medium

    International Nuclear Information System (INIS)

    Branzan, V.; Enache, N.

    1993-01-01

    The possibility of collectivization of an ensemble of atoms of an extended system (the distance between atoms is larger or equal to the wave-length of a spontaneous emitted radiation) during two-photon spontaneous decay is theoretically investigated. It is demonstrated that such systems of inverted atoms should emit phase-correlated pairs of photons. The time-space correlation among atoms is realized due to the two-photon exchanging through the electromagnetic field's vacuum. An increase of the spontaneous decay rate of the two-atom inverted ensemble is demonstrated. The dependence of two-photon superradiance on the sample geometry is investigated. A non-equilibrium method of the elimination of the atoms level Fermi-operators is proposed. (Author)

  3. A new approach to dual-color two-photon microscopy with fluorescent proteins

    Directory of Open Access Journals (Sweden)

    Rebane Aleks

    2010-02-01

    Full Text Available Abstract Background Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA efficiency. Results Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Conclusion Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.

  4. Folate receptor targeting silica nanoparticle probe for two-photon fluorescence bioimaging

    Science.gov (United States)

    Wang, Xuhua; Yao, Sheng; Ahn, Hyo-Yang; Zhang, Yuanwei; Bondar, Mykhailo V.; Torres, Joseph A.; Belfield, Kevin D.

    2010-01-01

    Narrow dispersity organically modified silica nanoparticles (SiNPs), diameter ~30 nm, entrapping a hydrophobic two-photon absorbing fluorenyl dye, were synthesized by hydrolysis of triethoxyvinylsilane and (3-aminopropyl)triethoxysilane in the nonpolar core of Aerosol-OT micelles. The surface of the SiNPs were functionalized with folic acid, to specifically deliver the probe to folate receptor (FR) over-expressing Hela cells, making these folate two-photon dye-doped SiNPs potential candidates as probes for two-photon fluorescence microscopy (2PFM) bioimaging. In vitro studies using FR over-expressing Hela cells and low FR expressing MG63 cells demonstrated specific cellular uptake of the functionalized nanoparticles. One-photon fluorescence microscopy (1PFM) imaging, 2PFM imaging, and two-photon fluorescence lifetime microscopy (2P-FLIM) imaging of Hela cells incubated with folate-modified two-photon dye-doped SiNPs were demonstrated. PMID:21258480

  5. Squeezing via two-photon transitions

    Science.gov (United States)

    Savage, C. M.; Walls, D. F.

    1986-05-01

    The squeezing spectrum for a cavity field mode interacting with an ensemble of three-level 'Lambda-configuration' atoms by an effective two-photon transition is calculated. The advantage of the three-level Lambda system as a squeezing medium, that is, optical nonlinearity without atomic saturation, has recently been pointed out by Reid, Walls, and Dalton. Perfect squeezing is predicted at the turning points for dispersive optical bistability and good squeezing for a range of other cases. Three-level ladder atoms interacting by an effective two-photon transition are also shown to give perfect squeezing in the dispersive limit.

  6. Two-photon excitation of argon

    International Nuclear Information System (INIS)

    Pindzola, P.S.; Payne, M.C.

    1982-01-01

    The authors calculate two photon excitation parameters for various excited states of argon assuming the absorption of near resonance broad-bandwidth laser radiation. Results are given for the case of two photons absorbed for the same laser beam as well as the case of absorbing photons of different frequency from each of two laser beams. The authors use multiconfiguration Hartree-Fock wave functions to evaluate the second-order sums over matrix elements. Various experimental laser schemes are suggested for the efficient excitation and subsequent ionization of argon

  7. A review of two photon physics

    International Nuclear Information System (INIS)

    Cooper, S.

    1982-08-01

    This talk is intended as an introduction for those not yet expert in two-photon physics, especially those e + e - one-photon physicists who still think of two-photon events as background. I concentrate on the physics questions involved, especially emphasizing the areas where I feel progress can be made in the near future, and of necessity leaving most experimental details to be found in the references. After a quick survey of the field and a few words about kinematics, I discuss in detail two major fields: the photon structure function and resonance production. (orig.)

  8. Review of Two-Photon Interactions

    International Nuclear Information System (INIS)

    Urner, David

    2004-01-01

    Presented are recent results of two-photon interactions. Topics include photon structure functions, inclusive hadron production, differential cross sections derived from tagged γγ fusion events and results in exclusive hadron production, particularly the observations of the η'c

  9. Two-photon transitions to exciton polaritons

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1979-08-01

    A semiclassical theory for the creation of excitonic polariton states by two-photon absorption, via an intermediate exciton state, is given. A band model has been introduced which gives the dominant contribution to this process. A numerical calculation is found to be in good agreement with a recent observation in CuCl. (author)

  10. Pion-pair production by two photons

    International Nuclear Information System (INIS)

    Terazawa, Hidezumi.

    1994-07-01

    The cross section for pion-pair production by two photons is calculated approximately by using the low energy theorem previously derived from partially-conserved-axial-vector-current hypothesis and current algebra, and found to agree very well with the experimental data recently obtained by the Mark II, TPC/Two-Gamma and CLEO Collaborations. (author)

  11. Optimization of a Time-Lapse Gravity Network for Carbon Sequestration

    Science.gov (United States)

    Appriou, D.; Strickland, C. E.; Ruprecht Yonkofski, C. M.

    2017-12-01

    The objective of this study is to evaluate what could be a comprehensive and optimal state of the art gravity monitoring network that would meet the UIC class VI regulation and insure that 90% of the CO2 injected remain underground. Time-lapse gravity surveys have a long history of effective applications of monitoring temporal density changes in the subsurface. For decades, gravity measurements have been used for a wide range of applications. The interest of time-lapse gravity surveys for monitoring carbon sequestration sites started recently. The success of their deployment in such sites depends upon a combination of favorable conditions, such as the reservoir geometry, depth, thickness, density change over time induced by the CO2 injection and the location of the instrument. In most cases, the density changes induced by the CO2 plume in the subsurface are not detectable from the surface but the use of borehole gravimeters can provide excellent results. In the framework of the National Assessment and Risk Partnership (NRAP) funded by the Department of Energy, the evaluation of the effectiveness of the gravity monitoring of a CO2 storage site has been assessed using multiple synthetic scenarios implemented on a community model developed for the Kimberlina site (e.g., fault leakage scenarios, borehole leakage). The Kimberlina carbon sequestration project was a pilot project located in southern San Joaquin Valley, California, aimed to safely inject 250,000 t CO2/yr for four years. Although the project was cancelled in 2012, the site characterization efforts resulted in the development of a geologic model. In this study, we present the results of the time-lapse gravity monitoring applied on different multiphase flow and reactive transport models developed by Lawrence Berkeley National Laboratory (i.e., no leakage, permeable fault zone, wellbore leakage). Our monitoring approach considers an ideal network, consisting of multiple vertical and horizontal instrumented

  12. The Relationship between Cell Number, Division Behavior and Developmental Potential of Cleavage Stage Human Embryos: A Time-Lapse Study.

    Directory of Open Access Journals (Sweden)

    Xiangyi Kong

    Full Text Available Day 3 cleavage embryo transfer is routine in many assisted reproductive technology centers today. Embryos are usually selected according to cell number, cell symmetry and fragmentation for transfer. Many studies have showed the relationship between cell number and embryo developmental potential. However, there is limited understanding of embryo division behavior and their association with embryo cell number and developmental potential. A retrospective and observational study was conducted to investigate how different division behaviors affect cell number and developmental potential of day 3 embryos by time-lapse imaging. Based on cell number at day 3, the embryos (from 104 IVF/intracytoplasmic sperm injection (ICSI treatment cycles, n = 799 were classified as follows: less than 5 cells (10C; n = 42. Division behavior, morphokinetic parameters and blastocyst formation rate were analyzed in 5 groups of day 3 embryos with different cell numbers. In 10C embryos increased compared to 7-8C embryos (45.8%, 33.3% vs. 11.1%, respectively. In ≥5C embryos, FR and DC significantly reduced developmental potential, whereas 10C. In NB embryos, the cell cycle elongation or shortening was the main cause for abnormally low or high cell number, respectively. After excluding embryos with abnormal division behaviors, the developmental potential, implantation rate and live birth rate of day 3 embryos increased with cell number.

  13. Two-Photon Excitation Microscopy for the Study of Living Cells and Tissues

    Science.gov (United States)

    Benninger, Richard K.P.; Piston, David W.

    2013-01-01

    Two-photon excitation microscopy is an alternative to confocal microscopy that provides advantages for three-dimensional and deep tissue imaging. This unit will describe the basic physical principles behind two-photon excitation and discuss the advantages and limitations of its use in laser-scanning microscopy. The principal advantages of two-photon microscopy are reduced phototoxicity, increased imaging depth, and the ability to initiate highly localized photochemistry in thick samples. Practical considerations for the application of two-photon microscopy will then be discussed, including recent technological advances. This unit will conclude with some recent applications of two-photon microscopy that highlight the key advantages over confocal microscopy and the types of experiments which would benefit most from its application. PMID:23728746

  14. In-vivo third-harmonic generation microscopy at 1550nm three-dimensional long-term time-lapse studies in living C. elegans embryos

    Science.gov (United States)

    Aviles-Espinosa, Rodrigo; Santos, Susana I. C. O.; Brodschelm, Andreas; Kaenders, Wilhelm G.; Alonso-Ortega, Cesar; Artigas, David; Loza-Alvarez, Pablo

    2011-03-01

    In-vivo microscopic long term time-lapse studies require controlled imaging conditions to preserve sample viability. Therefore it is crucial to meet specific exposure conditions as these may limit the applicability of established techniques. In this work we demonstrate the use of third harmonic generation (THG) microscopy for long term time-lapse three-dimensional studies (4D) in living Caenorhabditis elegans embryos employing a 1550 nm femtosecond fiber laser. We take advantage of the fact that THG only requires the existence of interfaces to generate signal or a change in the refractive index or in the χ3 nonlinear coefficient, therefore no markers are required. In addition, by using this wavelength the emitted THG signal is generated at visible wavelengths (516 nm) enabling the use of standard collection optics and detectors operating near their maximum efficiency. This enables the reduction of the incident light intensity at the sample plane allowing to image the sample for several hours. THG signal is obtained through all embryo development stages, providing different tissue/structure information. By means of control samples, we demonstrate that the expected water absorption at this wavelength does not severely compromise sample viability. Certainly, this technique reduces the complexity of sample preparation (i.e. genetic modification) required by established linear and nonlinear fluorescence based techniques. We demonstrate the non-invasiveness, reduced specimen interference, and strong potential of this particular wavelength to be used to perform long-term 4D recordings.

  15. Calibrating vadose zone models with time-lapse gravity data: a forced infiltration experiment

    DEFF Research Database (Denmark)

    Christiansen, Lars; Hansen, Allan Bo; Zibar, Majken Caroline Looms

    A change in soil water content is a change in mass stored in the subsurface, and when large enough, can be measured with a gravity meter. Over the last few decades there has been increased use of ground-based time-lapse gravity measurements to infer hydrogeological parameters. These studies have...... focused on the saturated zone, with specific yield as the most prominent target parameter and with few exceptions, changes in storage in the vadose zone have been considered as noise. Here modeling results are presented suggesting that gravity changes will be measureable when soil moisture changes occur...... in the unsaturated zone. These results are confirmed by field measurements of gravity and georadar data at a forced infiltration experiment conducted over 14 days on a grassland area of 10 m by 10 m. An unsaturated zone infiltration model can be calibrated using the gravity data with good agreement to the field data...

  16. Slow speed—fast motion: time-lapse recordings in physics education

    Science.gov (United States)

    Vollmer, Michael; Möllmann, Klaus-Peter

    2018-05-01

    Video analysis with a 30 Hz frame rate is the standard tool in physics education. The development of affordable high-speed-cameras has extended the capabilities of the tool for much smaller time scales to the 1 ms range, using frame rates of typically up to 1000 frames s-1, allowing us to study transient physics phenomena happening too fast for the naked eye. Here we want to extend the range of phenomena which may be studied by video analysis in the opposite direction by focusing on much longer time scales ranging from minutes, hours to many days or even months. We discuss this time-lapse method, needed equipment and give a few hints of how to produce respective recordings for two specific experiments.

  17. Using time-lapse gravity for groundwater model calibration: An application to alluvial aquifer storage

    DEFF Research Database (Denmark)

    Christiansen, Lars; Binning, Philip John; Rosbjerg, Dan

    2011-01-01

    hydrogeophysical inversion to decrease parameter correlation in groundwater models. This is demonstrated for a model of riverbank infiltration where combined inversion successfully constrains hydraulic conductivity and specific yield in both an analytical and a numerical groundwater model. A sensitivity study...... shows that time-lapse gravity data are especially useful to constrain specific yield. Furthermore, we demonstrate that evapotranspiration, and riverbed conductance are better constrained by coupled inversion to gravity and head data than to head data alone. When estimating the four parameters...... simultaneously, the six correlation coefficients were reduced from unity when only head data were employed to significantly lower values when gravity and head data were combined. Our analysis reveals that the estimated parameter values are not very sensitive to the choice of weighting between head and gravity...

  18. Time-lapse seismic analysis of the North Sea Fulmar Field

    Energy Technology Data Exchange (ETDEWEB)

    Johnston, David H.; McKenny, Robert S.; Burkhart, Tucker D.

    1998-12-31

    Time-lapse seismic analysis has been applied to two 3-D seismic surveys acquired over the central North Sea Fulmar field in a pre-production survey shot in 1977, reprocessed in 1987, and a survey in 1992. The Upper Jurassic reservoirs in the field have been under production since 1982. Differences in averaged impedance between the 1977 and 1992 surveys clearly show the effects of water influx and pressure decline. The changes observed in the seismic data are overall consistent with predictions obtained from a full-field, history-matched simulation. Differences in details may suggest areas of bypassed oil. Dta quality is not sufficient to serve as the sole basis for drilling decisions. 1 ref., 6 figs.

  19. Time-lapse cinematography of the capillary tube cell migration inhibition test.

    Science.gov (United States)

    Bray, M A

    1980-01-01

    The kinetics of human and guinea pig cell migration inhibition have been studied using time-lapse cinematography of cells migrating from capillary tubes. Guinea pig and human cells exhibit markedly different kinetics in the absence of inhibitors. Specific antigen causes a dose-related inhibition of migration for up to 60 h using guinea pig cells and a peak of inhibition after 18 h using the human leucocyte system. The timing of measurement of maximum activity more critical for the latter test. The kinetics of lymphokine generation have been examined and the migration inhibitory activity of the plant mitogen (PHA), a Kurloff cell product and a continuous cell line supernatant have been compared with the inhibitory profiles of lymphokine preparations and specific antigen.

  20. Time-lapse cinematography of dynamic changes occurring during in vitro development of human embryos.

    Science.gov (United States)

    Mio, Yasuyuki; Maeda, Kazuo

    2008-12-01

    The purpose of this study was to clarify developmental changes of early human embryos by using time-lapse cinematography (TLC). For human ova, fertilization and cleavage, development of the blastocyst, and hatching, as well as consequent changes were repeatedly photographed at intervals of 5-6 days by using an inverse microscope under stabilized temperature and pH. Photographs were taken at 30 frames per second and the movies were studied. Cinematography has increased our understanding of the morphologic mechanisms of fertilization, development, and behavior of early human embryos, and has identified the increased risk of monozygotic twin pregnancy based on prolonged incubation in vitro to the blastocyst stage. Using TLC, we observed the fertilization of an ovum by a single spermatozoon, followed by early cleavages, formation of the morula, blastocyst hatching, changes in the embryonic plates, and the development of monozygotic twins from the incubated blastocysts.

  1. Evaluating time-lapse ERT for monitoring DNAPL remediation via numerical simulation

    Science.gov (United States)

    Power, C.; Karaoulis, M.; Gerhard, J.; Tsourlos, P.; Giannopoulos, A.

    2012-12-01

    Dense non-aqueous phase liquids (DNAPLs) remain a challenging geoenvironmental problem in the near subsurface. Numerous thermal, chemical, and biological treatment methods are being applied at sites but without a non-destructive, rapid technique to map the evolution of DNAPL mass in space and time, the degree of remedial success is difficult to quantify. Electrical resistivity tomography (ERT) has long been presented as highly promising in this context but has not yet become a practitioner's tool due to challenges in interpreting the survey results at real sites where the initial condition (DNAPL mass, DNAPL distribution, subsurface heterogeneity) is typically unknown. Recently, a new numerical model was presented that couples DNAPL and ERT simulation at the field scale, providing a tool for optimizing ERT application and interpretation at DNAPL sites (Power et al., 2011, Fall AGU, H31D-1191). The objective of this study is to employ this tool to evaluate the effectiveness of time-lapse ERT to monitor DNAPL source zone remediation, taking advantage of new inversion methodologies that exploit the differences in the target over time. Several three-dimensional releases of chlorinated solvent DNAPLs into heterogeneous clayey sand at the field scale were generated, varying in the depth and complexity of the source zone (target). Over time, dissolution of the DNAPL in groundwater was simulated with simultaneous mapping via periodic ERT surveys. Both surface and borehole ERT surveys were conducted for comparison purposes. The latest four-dimensional ERT inversion algorithms were employed to generate time-lapse isosurfaces of the DNAPL source zone for all cases. This methodology provided a qualitative assessment of the ability of ERT to track DNAPL mass removal for complex source zones in realistically heterogeneous environments. In addition, it provided a quantitative comparison between the actual DNAPL mass removed and that interpreted by ERT as a function of depth below

  2. High-resolution three-dimensional imaging of a depleted CMOS sensor using an edge Transient Current Technique based on the Two Photon Absorption process (TPA-eTCT)

    CERN Document Server

    García, Marcos Fernández; Echeverría, Richard Jaramillo; Moll, Michael; Santos, Raúl Montero; Moya, David; Pinto, Rogelio Palomo; Vila, Iván

    2016-01-01

    For the first time, the deep n-well (DNW) depletion space of a High Voltage CMOS sensor has been characterized using a Transient Current Technique based on the simultaneous absorption of two photons. This novel approach has allowed to resolve the DNW implant boundaries and therefore to accurately determine the real depleted volume and the effective doping concentration of the substrate. The unprecedented spatial resolution of this new method comes from the fact that measurable free carrier generation in two photon mode only occurs in a micrometric scale voxel around the focus of the beam. Real three-dimensional spatial resolution is achieved by scanning the beam focus within the sample.

  3. High-resolution three-dimensional imaging of a depleted CMOS sensor using an edge Transient Current Technique based on the Two Photon Absorption process (TPA-eTCT)

    Energy Technology Data Exchange (ETDEWEB)

    García, Marcos Fernández; Sánchez, Javier González; Echeverría, Richard Jaramillo [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain); Moll, Michael [CERN, Organisation europénne pour la recherche nucléaire, CH-1211 Genéve 23 (Switzerland); Santos, Raúl Montero [SGIker Laser Facility, UPV/EHU, Sarriena, s/n - 48940 Leioa-Bizkaia (Spain); Moya, David [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain); Pinto, Rogelio Palomo [Departamento de Ingeniería Electrónica, Escuela Superior de Ingenieros Universidad de Sevilla (Spain); Vila, Iván [Instituto de Física de Cantabria (CSIC-UC), Avda. los Castros s/n, E-39005 Santander (Spain)

    2017-02-11

    For the first time, the deep n-well (DNW) depletion space of a High Voltage CMOS sensor has been characterized using a Transient Current Technique based on the simultaneous absorption of two photons. This novel approach has allowed to resolve the DNW implant boundaries and therefore to accurately determine the real depleted volume and the effective doping concentration of the substrate. The unprecedented spatial resolution of this new method comes from the fact that measurable free carrier generation in two photon mode only occurs in a micrometric scale voxel around the focus of the beam. Real three-dimensional spatial resolution is achieved by scanning the beam focus within the sample.

  4. Can Two-Photon Interference be Considered the Interference of Two Photons?

    International Nuclear Information System (INIS)

    Pittman, T.B.; Strekalov, D.V.; Migdall, A.; Rubin, M.H.; Sergienko, A.V.; Shih, Y.H.

    1996-01-01

    We report on a open-quote open-quote postponed compensation close-quote close-quote experiment in which the observed two-photon entangled state interference cannot be pictured in terms of the overlap of the two individual photon wave packets of a parametric down-conversion pair on a beam splitter. In the sense of a quantum eraser, the distinguishability of the different two-photon Feynman amplitudes leading to a coincidence detection is removed by delaying the compensation until after the output of an unbalanced two-photon interferometer. copyright 1996 The American Physical Society

  5. MONITORING EROSION OF STONE SURFACES USING TIME-LAPSE AND PTM PHOTOGRAPHY: FIELD STUDY OF A 14TH CENTURY MONASTERY IN YORKSHIRE

    Science.gov (United States)

    Doehne, E.; Pinchin, S.

    2009-12-01

    Evaluating stone weathering rates and their relationship to environmental fluctuations is an important challenge in understanding the critical zone and also in efforts to prevent the loss of important cultural heritage in stone, such as monuments, sculpture and archaeological sites. Repeat photography has been widely used to evaluate geological processes such as the retreat of glaciers and the weathering of stone surfaces. However, a fundamental difficulty is that the images are often shot under differing lighting conditions, making the interpretation of stone surface loss particularly challenging. Two developments in photographic documentation show promise for improving the situation. One is the use of digital time-lapse methods to provide more frequent images to correlate stone surface loss with ongoing environmental changes. The other is a relatively new method known as polynomial transform mapping (PTM), which integrates multiple photographs taken at different angles to document more comprehensively the texture of stone surfaces. Using Java-based software, the viewer can control the precise angle of the light source in an interpolated, high-quality image. PTM can produce raking light images from any angle, as well as images with ‘normal’ illumination. We present here results based on several years of macro-photography, time-lapse imaging, and PTM imaging of rapidly eroding stone surfaces at the site of Howden Minster in Yorkshire, UK, which suffers from salt weathering. The images show that surface loss is episodic rather than continuous and in some cases is related to unusual environmental conditions, such as high winds and condensation events. Damage was also found to be synchronous, with surface change (flaking, granular disintegration, and loss of flakes) occurring at the same time in different stone blocks. Crystallization pressure from phase transitions in magnesium sulfate salts appears to be the main cause of the loss of stone surfaces.

  6. Probing two-photon exchange with OLYMPUS

    International Nuclear Information System (INIS)

    Kohl, M.

    2014-01-01

    Two-photon exchange is believed to be responsible for the discrepancies in the proton electric to magnetic form factor ratio found with the Rosenbluth and polarization transfer methods. If this explanation is correct, one expects significant differences in the lepton-proton cross sections between positrons and electrons. The OLYMPUS experiment at DESY in Hamburg, Germany was designed to measure the ratio of unpolarized positron-proton and electron-proton elastic scattering cross sections over a wide kinematic range with high precision, in order to quantify the effect of two-photon exchange. The experiment used intense beams of electrons and positrons stored in the DORIS ring at 2.0 GeV interacting with an internal windowless hydrogen gas target. The current status of OLYMPUS will be discussed. (authors)

  7. Bayesian Markov chain Monte Carlo Inversion of Time-Lapse Geophysical Data To Characterize the Vadose Zone

    DEFF Research Database (Denmark)

    Scholer, Marie; Irving, James; Zibar, Majken Caroline Looms

    Geophysical methods have the potential to provide valuable information on hydrological properties in the unsaturated zone. In particular, time-lapse geophysical data, when coupled with a hydrological model and inverted stochastically, may allow for the effective estimation of subsurface hydraulic...... parameters and their corresponding uncertainties. In this study, we use a Bayesian Markov-chain-Monte-Carlo (MCMC) inversion approach to investigate how much information regarding vadose zone hydraulic properties can be retrieved from time-lapse crosshole GPR data collected at the Arrenaes field site...

  8. Symmetry adaptation in two-photon spectroscopy

    International Nuclear Information System (INIS)

    Kibler, M.

    1991-11-01

    Symmetry adaptation techniques are applied to the determination of the intensity of two-photon transitions for transition ions in finite symmetry environments. The case of intra-configurational transitions are discussed with some details and some results on inter-configurational transitions are briefly reported. In particular, for intra-configurational transitions, a model is described which takes into account the following ingredients: (symmetry, second- plus third-order mechanisms, S-, L- and J-mixings). (author) 20 refs

  9. Wide-field two-photon microscopy with temporal focusing and HiLo background rejection

    Science.gov (United States)

    Yew, Elijah Y. S.; Choi, Heejin; Kim, Daekeun; So, Peter T. C.

    2011-03-01

    Scanningless depth-resolved microscopy is achieved through spatial-temporal focusing and has been demonstrated previously. The advantage of this method is that a large area may be imaged without scanning resulting in higher throughput of the imaging system. Because it is a widefield technique, the optical sectioning effect is considerably poorer than with conventional spatial focusing two-photon microscopy. Here we propose wide-field two-photon microscopy based on spatio-temporal focusing and employing background rejection based on the HiLo microscope principle. We demonstrate the effects of applying HiLo microscopy to widefield temporally focused two-photon microscopy.

  10. Two-photon spin generation and detection

    Energy Technology Data Exchange (ETDEWEB)

    Miah, M Idrish, E-mail: m.miah@griffith.edu.a [Nanoscale Science and Technology Centre, Griffith University, Nathan, Brisbane, QLD 4111 (Australia)

    2009-02-21

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay ({Delta}t), lattice temperature (T{sub L}), doping density (n) as well as of the excess photon energy {Delta}E{sub 2{omega}}= {h_bar}2{omega} - E{sub g}, where E{sub g} is the band gap energy. P is found to be decayed with {Delta}t and enhanced with the decrease in T{sub L} or the increase in n. It is also found that P decreases with the increase in {Delta}E{sub 2{omega}}and depolarizes rapidly for {Delta}E{sub 2{omega}}> {Delta}E{sub SO}, where {Delta}E{sub SO} is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  11. Two-photon spin generation and detection

    International Nuclear Information System (INIS)

    Miah, M Idrish

    2009-01-01

    A time- and polarization-resolved two-photon pump-probe investigation is performed in lightly doped GaAs. We generate spin-polarized electrons in bulk GaAs at various temperatures using right-circularly polarized two-photon excitation and detect them by probing the spin-dependent transmission of the sample. The spin polarization (P) of conduction band electrons, as measured using probe pulses with the same (right) and opposite (left) circular polarization, is measured in dependences of pump-probe delay (Δt), lattice temperature (T L ), doping density (n) as well as of the excess photon energy ΔE 2ω = ℎ2ω - E g , where E g is the band gap energy. P is found to be decayed with Δt and enhanced with the decrease in T L or the increase in n. It is also found that P decreases with the increase in ΔE 2ω and depolarizes rapidly for ΔE 2ω > ΔE SO , where ΔE SO is the spin-orbit splitting energy. The results demonstrate that due to a much longer absorption depth highly polarized spins can be generated optically by two-photon pumping of bulk semiconductors.

  12. Two-photon excited fluorescence emission from hemoglobin

    Science.gov (United States)

    Sun, Qiqi; Zeng, Yan; Zhang, Wei; Zheng, Wei; Luo, Yi; Qu, Jianan Y.

    2015-03-01

    Hemoglobin, one of the most important proteins in blood, is responsible for oxygen transportation in almost all vertebrates. Recently, we discovered two-photon excited hemoglobin fluorescence and achieved label-free microvascular imaging based on the hemoglobin fluorescence. However, the mechanism of its fluorescence emission still remains unknown. In this work, we studied the two-photon excited fluorescence properties of the hemoglobin subunits, heme/hemin (iron (II)/(III) protoporphyrin IX) and globin. We first studied the properties of heme and the similar spectral and temporal characteristics of heme and hemoglobin fluorescence provide strong evidence that heme is the fluorophore in hemoglobin. Then we studied the fluorescence properties of hemin, globin and methemoglobin, and found that the hemin may have the main effect on the methemoglobin fluorescence and that globin has tryptophan fluorescence like other proteins. Finally, since heme is a centrosymmetric molecule, that the Soret band fluorescence of heme and hemoglobin was not observed in the single photon process in the previous study may be due to the parity selection rule. The discovery of heme two-photon excited fluorescence may open a new window for heme biology research, since heme as a cofactor of hemoprotein has many functions, including chemical catalysis, electron transfer and diatomic gases transportation.

  13. Two-Photon Fluorescence Microscopy Developed for Microgravity Fluid Physics

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2004-01-01

    Recent research efforts within the Microgravity Fluid Physics Branch of the NASA Glenn Research Center have necessitated the development of a microscope capable of high-resolution, three-dimensional imaging of intracellular structure and tissue morphology. Standard optical microscopy works well for thin samples, but it does not allow the imaging of thick samples because of severe degradation caused by out-of-focus object structure. Confocal microscopy, which is a laser-based scanning microscopy, provides improved three-dimensional imaging and true optical sectioning by excluding the out-of-focus light. However, in confocal microscopy, out-of-focus object structure is still illuminated by the incoming beam, which can lead to substantial photo-bleaching. In addition, confocal microscopy is plagued by limited penetration depth, signal loss due to the presence of a confocal pinhole, and the possibility of live-cell damage. Two-photon microscopy is a novel form of laser-based scanning microscopy that allows three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon microscopy, it utilizes the nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption because of the nonlinear (i.e., quadratic) electric field dependence, so an ultrafast pulsed laser source must typically be employed. On the other hand, this stringent energy density requirement effectively localizes fluorophore excitation to the focal volume. Consequently, two-photon microscopy provides optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction in photo-damage because of the longer excitation wavelength, a reduction in background fluorescence, and a 4 increase in penetration depth over confocal methods because of the reduction in Rayleigh scattering.

  14. Two-Photon Fluorescence Microscope for Microgravity Research

    Science.gov (United States)

    Fischer, David G.; Zimmerli, Gregory A.; Asipauskas, Marius

    2005-01-01

    A two-photon fluorescence microscope has been developed for the study of biophysical phenomena. Two-photon microscopy is a novel form of laser-based scanning microscopy that enables three-dimensional imaging without many of the problems inherent in confocal microscopy. Unlike one-photon optical microscopy, two-photon microscopy utilizes the simultaneous nonlinear absorption of two near-infrared photons. However, the efficiency of two-photon absorption is much lower than that of one-photon absorption, so an ultra-fast pulsed laser source is typically employed. On the other hand, the critical energy threshold for two-photon absorption leads to fluorophore excitation that is intrinsically localized to the focal volume. Consequently, two-photon microscopy enables optical sectioning and confocal performance without the need for a signal-limiting pinhole. In addition, there is a reduction (relative to one-photon optical microscopy) in photon-induced damage because of the longer excitation wavelength. This reduction is especially advantageous for in vivo studies. Relative to confocal microscopy, there is also a reduction in background fluorescence, and, because of a reduction in Rayleigh scattering, there is a 4 increase of penetration depth. The prohibitive cost of a commercial two-photon fluorescence-microscope system, as well as a need for modularity, has led to the construction of a custom-built system (see Figure 1). This system includes a coherent mode-locked titanium: sapphire laser emitting 120-fs-duration pulses at a repetition rate of 80 MHz. The pulsed laser has an average output power of 800 mW and a wavelength tuning range of 700 to 980 nm, enabling the excitation of a variety of targeted fluorophores. The output from the laser is attenuated, spatially filtered, and then directed into a confocal scanning head that has been modified to provide for side entry of the laser beam. The laser output coupler has been replaced with a dichroic filter that reflects the

  15. Micromechanical Time-Lapse X-ray CT Study of Fatigue Damage in Uni-Directional Fibre Composites

    DEFF Research Database (Denmark)

    Jespersen, Kristine Munk; Lowe, Tristan; Withers, Philip J.

    2015-01-01

    . The geometry of the cut-out is similar to that which will be used in the time-lapse study. As the micro-mechanical damage mechanisms are small features, it is necessary to obtain a high scan resolution which sets a limit to how large the field of view can be. Therefore, it is necessary to perform several scans...

  16. Land time-lapse CSEM : Collecting, modeling and inversion of CSEM data for a steam-injected oil field

    NARCIS (Netherlands)

    Schaller, A.M.

    2018-01-01

    Geophysical methods are widely used for hydrocarbon exploration and time-lapse measurements. One method that can be applied in place of or in addition to the routinely used seismic method, is the Controlled-Source ElectroMagnetic (CSEM) method. The work described in this thesis explores various

  17. Time-lapse micro-tomography analysis of the deformation response of a gellan-gum-based scaffold

    Czech Academy of Sciences Publication Activity Database

    Kytýř, Daniel; Fenclová, Nela; Zlámal, Petr; Kumpová, Ivana; Fíla, Tomáš; Koudelka_ml., Petr; Gantar, A.; Novak, S.

    2017-01-01

    Roč. 51, č. 3 (2017), s. 397-402 ISSN 1580-2949 Institutional support: RVO:68378297 Keywords : bone scaffold * gellan gum * time-lapse micro CT * digital volume correlation Subject RIV: JJ - Other Materials OBOR OECD: Materials engineering Impact factor: 0.436, year: 2016 http://mit.imt.si/Revija/izvodi/mit173/kytyr.pdf

  18. Mass conservative three-dimensional water tracer distribution from MCMC inversion of time-lapse GPR data

    NARCIS (Netherlands)

    Lalov, E.; Linde, N.; Vrugt, J.A.

    2012-01-01

    Time-lapse geophysical measurements are widely used to monitor the movement of water and solutes through the subsurface. Yet commonly used deterministic least squares inversions typically suffer from relatively poor mass recovery, spread overestimation, and limited ability to appropriately estimate

  19. Joint inversion of time-lapse VSP data for monitoring CO2 injection at the Farnsworth EOR field in Texas

    Science.gov (United States)

    Zhang, M.; Gao, K.; Balch, R. S.; Huang, L.

    2016-12-01

    During the Development Phase (Phase III) of the U.S. Southwest Regional Partnership on Carbon Sequestration (SWP), time-lapse 3D vertical seismic profiling (VSP) data were acquired to monitor CO2 injection/migration at the Farnsworth Enhanced Oil Recovery (EOR) field, in partnership with the industrial partner Chaparral Energy. The project is to inject a million tons of carbon dioxide into the target formation, the deep oil-bearing Morrow Formation in the Farnsworth Unit EOR field. Quantitative time-lapse seismic monitoring has the potential to track CO2 movement in geologic carbon storage sites. Los Alamos National Laboratory (LANL) has recently developed new full-waveform inversion methods to jointly invert time-lapse seismic data for changes in elastic and anisotropic parameters in target monitoring regions such as a CO2 reservoir. We apply our new joint inversion methods to time-lapse VSP data acquired at the Farnsworth EOR filed, and present some preliminary results showing geophysical properties changes in the reservoir.

  20. Time-Lapse Analysis of Methane Quantity in the Mary Lee Group of Coal Seams Using Filter-Based Multiple-Point Geostatistical Simulation.

    Science.gov (United States)

    Karacan, C Özgen; Olea, Ricardo A

    2013-08-01

    Coal seam degasification and its success are important for controlling methane, and thus for the health and safety of coal miners. During the course of degasification, properties of coal seams change. Thus, the changes in coal reservoir conditions and in-place gas content as well as methane emission potential into mines should be evaluated by examining time-dependent changes and the presence of major heterogeneities and geological discontinuities in the field. In this work, time-lapsed reservoir and fluid storage properties of the New Castle coal seam, Mary Lee/Blue Creek seam, and Jagger seam of Black Warrior Basin, Alabama, were determined from gas and water production history matching and production forecasting of vertical degasification wellbores. These properties were combined with isotherm and other important data to compute gas-in-place (GIP) and its change with time at borehole locations. Time-lapsed training images (TIs) of GIP and GIP difference corresponding to each coal and date were generated by using these point-wise data and Voronoi decomposition on the TI grid, which included faults as discontinuities for expansion of Voronoi regions. Filter-based multiple-point geostatistical simulations, which were preferred in this study due to anisotropies and discontinuities in the area, were used to predict time-lapsed GIP distributions within the study area. Performed simulations were used for mapping spatial time-lapsed methane quantities as well as their uncertainties within the study area. The systematic approach presented in this paper is the first time in literature that history matching, TIs of GIPs and filter simulations are used for degasification performance evaluation and for assessing GIP for mining safety. Results from this study showed that using production history matching of coalbed methane wells to determine time-lapsed reservoir data could be used to compute spatial GIP and representative GIP TIs generated through Voronoi decomposition

  1. Radiative corrections to two photon physics

    International Nuclear Information System (INIS)

    Neervan, W.L. van; Vermaseren, J.A.M.

    1983-06-01

    The authors develop a method to calculate radiative corrections to two photon reactions of the type e + e - →e + e - X where X is an arbitrary final state. To illustrate this they take the example where X stands for a point-like pseudoscalar. It will be shown that the method is an improvement on the standard way in adding real and virtual photon contributions to the (differential) cross-sections. This makes it possible to compute differential and total cross-sections to a very high precision and with a minimum of computer time which is not possible by using more conventional methods. (Auth.)

  2. Inclusive two-photon reactions at TRISTAN

    International Nuclear Information System (INIS)

    Drees, M.

    1995-01-01

    After briefly reviewing past accomplishments of TRISTAN experiments in the field of inclusive two-photon reactions, I discuss open problems in the Monte Carlo simulation of such reactions. The main emphasis is on multiple scattering, i.e. events where at least two pairs of partons scatter within the same γγ collision to form at least four (mini)jets. The cross section for such events might just be observable at TRISTAN. While theoretical arguments for the existence of such events are strong, they have not yet been directly observed experimentally, thereby potentially opening a new opportunity for TRISTAN experiments. (author)

  3. Two-photon physics at LEP

    International Nuclear Information System (INIS)

    Ginzburg, I.F.

    1988-01-01

    The two-photon production of hadrons in e + e - collisions e + e - →e + e - h from which the γγ→h cross sections are extracted is discussed. The common features of these processes are: hadrons move, as a rule, along e + e - beam axis, their total transverse momentum K perpendicular or perpendicular to is small; the total hadron energy is usually less than √S/2. Physical problems of soft processes, exotics, hard processes, semihard processes are considered. New possibilities of LEP, the most interesting and real are presented

  4. Two-photon cooling of magnesium atoms

    DEFF Research Database (Denmark)

    Malossi, N.; Damkjær, S.; Hansen, P. L.

    2005-01-01

    A two-photon mechanism for cooling atoms below the Doppler temperature is analyzed. We consider the magnesium ladder system (3s2)S01¿(3s3p)P11 at 285.2nm followed by the (3s3p)P11¿(3s3d)D21 transition at 880.7nm . For the ladder system quantum coherence effects may become important. Combined with...... and experiment is excellent. In addition, by properly choosing the Rabi frequencies of the two optical transitions a velocity independent atomic dark state is observed....

  5. Thermal erosion of a permafrost coastline: Improving process-based models using time-lapse photography

    Science.gov (United States)

    Wobus, C.; Anderson, R.; Overeem, I.; Matell, N.; Clow, G.; Urban, F.

    2011-01-01

    Coastal erosion rates locally exceeding 30 m y-1 have been documented along Alaska's Beaufort Sea coastline, and a number of studies suggest that these erosion rates have accelerated as a result of climate change. However, a lack of direct observational evidence has limited our progress in quantifying the specific processes that connect climate change to coastal erosion rates in the Arctic. In particular, while longer ice-free periods are likely to lead to both warmer surface waters and longer fetch, the relative roles of thermal and mechanical (wave) erosion in driving coastal retreat have not been comprehensively quantified. We focus on a permafrost coastline in the northern National Petroleum Reserve-Alaska (NPR-A), where coastal erosion rates have averaged 10-15 m y-1 over two years of direct monitoring. We take advantage of these extraordinary rates of coastal erosion to observe and quantify coastal erosion directly via time-lapse photography in combination with meteorological observations. Our observations indicate that the erosion of these bluffs is largely thermally driven, but that surface winds play a crucial role in exposing the frozen bluffs to the radiatively warmed seawater that drives melting of interstitial ice. To first order, erosion in this setting can be modeled using formulations developed to describe iceberg deterioration in the open ocean. These simple models provide a conceptual framework for evaluating how climate-induced changes in thermal and wave energy might influence future erosion rates in this setting.

  6. Monitoring of In-Situ Remediation By Time Lapse 3D Geo-Electric Measurements

    Science.gov (United States)

    Kanli, A. I.; Tildy, P.; Neducza, B.; Nagy, P.; Hegymegi, C.

    2017-12-01

    Injection of chemical oxidant solution to degrade the subsurface contaminants can be used for hydrocarbon contamination remediation. In this study, we developed a non-destructive measurement strategy to monitor oxidative in-situ remediation processes. The difficulties of the presented study originate from the small volume of conductive solution that can be used due to environmental considerations. Due to the effect of conductive groundwater and the high clay content of the targeted layer and the small volume of conductive solution that can be used due to environmental considerations, a site specific synthetic modelling is necessary for measurement design involving the results of preliminary 2D ERT measurements, electrical conductivity measurements of different active agents and expected resistivity changes calculated by soil resistivity modelling. Because of chemical biodegradation, the results of soil resistivity modelling have suggested that the reagent have complex effects on contaminated soils. As a result the plume of resistivity changes caused by the injected agent was determined showing strong fracturing effect because of the high pressure of injection. 3D time-lapse geo-electric measurements were proven to provide a usable monitoring tool for in-situ remediation as a result of our sophisticated tests and synthetic modelling.

  7. Time-lapse analysis of potential cellular responsiveness to Johrei, a Japanese healing technique

    Directory of Open Access Journals (Sweden)

    Moore Dan

    2005-01-01

    Full Text Available Abstract Background Johrei is an alternative healing practice which involves the channeling of a purported universal healing energy to influence the health of another person. Despite little evidence to support the efficacy of such practices the use of such treatments is on the rise. Methods We assessed cultured human cancer cells for potential responsiveness to Johrei treatment from a short distance. Johrei treatment was delivered by practitioners who participated in teams of two, alternating every half hour for a total of four hours of treatment. The practitioners followed a defined set of mental procedures to minimize variability in mental states between experiments. An environmental chamber maintained optimal growth conditions for cells throughout the experiments. Computerized time-lapse microscopy allowed documentation of cancer cell proliferation and cell death before, during and after Johrei treatments. Results Comparing eight control experiments with eight Johrei intervention experiments, we found no evidence of a reproducible cellular response to Johrei treatment. Conclusion Cell death and proliferation rates of cultured human cancer cells do not appear responsive to Johrei treatment from a short distance.

  8. Monitoring the snowpack volume in a sinkhole on Mount Lebanon using time lapse Photogrammetry

    Science.gov (United States)

    Abou Chakra, C.; Gascoin, S.; Somma, J.; Drapeau, L.; Fanise, P.

    2017-12-01

    Lebanon is one of the richest country in the Middle East for water resources, thanks to its mountain ranges that trigger precipitation from the moist air masses coming from the Mediterranean Sea. Snowpack acts as natural water storage in winter and supply fresh water during spring and summer. Yet, Lebanon is facing a serious water scarcity problem due to: i) decreasing amount of precipitation and climate change; ii) major growth of population of original residence and large number of refugees during regional wars. Therefore, continuous and systematic monitoring of the Lebanese water resources is becoming crucial. The Mount Lebanon is made of karstic depressions named "sinkholes". It is important to monitor the snowmelt process inside these sinkholes because of their key role as "containers" of seasonal snow. By isolating the snowpack from sun radiation and wind, they slow down the natural melting process and sublimation, thus delaying as well the low water flow period. An observatory is set up to monitor the snowpack evolution in a pilot sinkhole located in Mount Lebanon. The system uses three time-lapse cameras and structure-from-motion principles to reconstruct the snow volume within the sinkhole. The approach is validated by standard topographic surveys. The results indicate that snow depth can be retrieved with an accuracy between 20 and 60 cm (residuals standard deviation) and a low bias of 50 cm after coregistration of the digital elevation models.

  9. Direct prediction of spatially and temporally varying physical properties from time-lapse electrical resistance data

    Science.gov (United States)

    Hermans, Thomas; Oware, Erasmus; Caers, Jef

    2016-09-01

    Time-lapse applications of electrical methods have grown significantly over the last decade. However, the quantitative interpretation of tomograms in terms of physical properties, such as salinity, temperature or saturation, remains difficult. In many applications, geophysical models are transformed into hydrological models, but this transformation suffers from spatially and temporally varying resolution resulting from the regularization used by the deterministic inversion. In this study, we investigate a prediction-focused approach (PFA) to directly estimate subsurface physical properties with electrical resistance data, circumventing the need for classic tomographic inversions. First, we generate a prior set of resistance data and physical property forecast through hydrogeological and geophysical simulations mimicking the field experiment. We reduce the dimension of both the data and the forecast through principal component analysis in order to keep the most informative part of both sets in a reduced dimension space. Then, we apply canonical correlation analysis to explore the relationship between the data and the forecast in their reduced dimension space. If a linear relationship can be established, the posterior distribution of the forecast can be directly sampled using a Gaussian process regression where the field data scores are the conditioning data. In this paper, we demonstrate PFA for various physical property distributions. We also develop a framework to propagate the estimated noise level in the reduced dimension space. We validate the results by a Monte Carlo study on the posterior distribution and demonstrate that PFA yields accurate uncertainty for the cases studied.

  10. A clustering approach applied to time-lapse ERT interpretation - Case study of Lascaux cave

    Science.gov (United States)

    Xu, Shan; Sirieix, Colette; Riss, Joëlle; Malaurent, Philippe

    2017-09-01

    The Lascaux cave, located in southwest France, is one of the most important prehistoric cave in the world that shows Paleolithic paintings. This study aims to characterize the structure of the weathered epikarst setting located above the cave using Time-Lapse Electrical Resistivity Tomography (ERT) combined with local hydrogeological and climatic environmental data. Twenty ERT profiles were carried out for two years and helped us to record the seasonal and spatial variations of the electrical resistivity of the hydraulic upstream area of the Lascaux cave. The 20 interpreted resistivity models were merged into a single synthetic model using a multidimensional statistical method (Hierarchical Agglomerative Clustering). The individual blocks from the synthetic model associated with a similar resistivity variability were gathered into 7 clusters. We combined the resistivity temporal variations with climatic and hydrogeological data to propose a geo-electrical model that relates to a conceptual geological model. We provide a geological interpretation for each cluster regarding epikarst features. The superficial clusters (no 1 & 2) are linked to effective rainfall and trees, probably a fractured limestone. Another two clusters (no 6 & 7) are linked to detrital formations (sand and clay respectively). The cluster 3 may correspond to a marly limestone that forms a non-permeable horizon. Finally, the electrical behavior of the last two clusters (no 4 & 5) is correlated with the variation of flow rate; they may be a privileged feed zone of the flow in the cave.

  11. Environmental Monitoring Of Leaks Using Time Lapsed Long Electrode Electrical Resistivity

    International Nuclear Information System (INIS)

    Rucker, D.F.; Fink, J.B.; Loke, M.H.; Myers, D.A.

    2009-01-01

    Highly industrialized areas pose significant challenges for surface based electrical resistivity characterization and monitoring due to the high degree of metallic infrastructure. The infrastructure is typically several orders of magnitude more conductive than the desired targets, preventing the geophysicist from obtaining a clear picture of the subsurface. These challenges may be minimized if steel-cased wells are used as long electrodes. We demonstrate a method of using long electrodes in a complex nuclear waste facility to monitor a simulated leak from an underground storage tank. The leak was simulated by injecting high conductivity fluid in a perforated well and the resistivity measurements were made before and after the leak test. The data were processed in four dimensions, where a regularization procedure was applied in both the time and space domains. The results showed a lowered resistivity feature develop south of the injection site. The time lapsed regularization parameter had a strong influence on the differences in inverted resistivity between the pre and post datasets, potentially making calibration of the results to specific hydrogeologic parameters difficult.

  12. Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography.

    Science.gov (United States)

    Ugajin, Tomohisa; Terada, Yukihiro; Hasegawa, Hisataka; Velayo, Clarissa L; Nabeshima, Hiroshi; Yaegashi, Nobuo

    2010-05-15

    To analyze whether blastomere biopsy affects early embryonal growth as observed through time-lapse cinematography. Comparative prospective study between embryos in which a blastomere was removed and embryos in which a blastomere was not removed. An experimental laboratory of the university. We calculated the time between blastocele formation and the end of hatching, the time between the start and end of hatching, the number of contractions and expansions between blastocyst formation and the end of hatching, and the maximum diameter of the expanded blastocyst. In blastomere removal embryos, compaction began at the six-cell stage instead of at the eight-cell stage. We also found that hatching was delayed in these embryos as compared with matched controls. Moreover, the frequency of contraction and expansion movements after blastocyst formation was significantly higher in the blastomere removal group as compared with the control group. Finally, the maximum diameter of the expanded blastocyst just before hatching was not significantly different between both groups. These findings suggested that blastomere removal has an adverse effect on embryonic development around the time of hatching. Thus, future developments in preimplantation genetic diagnosis and screening should involve further consideration and caution in light of the influence of blastomere biopsy on embryonal growth. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. Mathematical analysis of endothelial sibling pair cell-cell interactions using time-lapse cinematography data.

    Science.gov (United States)

    Brown, L M; Ryan, U S; Absher, M; Olazabal, B M

    1982-01-01

    The sibling pairs from two different endothelial cell cultures were analysed by time-lapse cinematography. It was shown that wounded and regular (low density seeded) cultures differed in the behaviour patterns of their siblings. The cultures differed most significantly in the minimum interdivision time (IDT) which was 27% lower for the wounded culture. In the wounded culture there was a greater correlation of IDT values between sibling pairs. IDT values recorded both for paired and for unpaired cells were shorter for the wounded than for the regular culture. The mean IDT for unpaired cells was longer than the mean IDT for paired cells in the regular culture. Thus paired cells in the regular culture, had shorter IDTs, but not as short as in the wounded culture. It was significant that in the wounded culture the first generation of siblings were very close (less than 150 microns apart) at division. Overall the behaviour differences between the two cultures resulted in a higher rate of increase in cell numbers, and thus faster repair, of the wounded monolayer.

  14. A series of fluorene-based two-photon absorbing molecules: synthesis, linear and nonlinear characterization, and bioimaging

    Science.gov (United States)

    Andrade, Carolina D.; Yanez, Ciceron O.; Rodriguez, Luis; Belfield, Kevin D.

    2010-01-01

    The synthesis, structural, and photophysical characterization of a series of new fluorescent donor–acceptor and acceptor-acceptor molecules, based on the fluorenyl ring system, with two-photon absorbing properties is described. These new compounds exhibited large Stokes shifts, high fluorescent quantum yields, and, significantly, high two-photon absorption cross sections, making them well suited for two-photon fluorescence microscopy (2PFM) imaging. Confocal and two-photon fluorescence microscopy imaging of COS-7 and HCT 116 cells incubated with probe I showed endosomal selectivity, demonstrating the potential of this class of fluorescent probes in multiphoton fluorescence microscopy. PMID:20481596

  15. Resonant production in two photon collisions

    International Nuclear Information System (INIS)

    Butler, F.

    1988-12-01

    Using 220 picobarn/sup /minus/1/ of data collected by the Mark II detector at PEP, we have measured the width of the /eta/' for quasi real photons. The width is separately measured in the three reactions e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → γγ, e + e/sup /minus// → e + e/sup /minus///eta/ 0 π + π/sup /minus//; /eta/ 0 → π + π/sup /minus//(π 0 ,γ) and e + e/sup /minus// → e + e/sup /minus///rho/ 0 γ, giving a statistically weighted average two-photon width of Γ/sub /eta/'→γγ/ = 5. 21+-0.28 keV. As a by-product of the measurement of Γ/sub /eta/'→γγ/ using the decay mode /eta/' → /rho/γ, we also measured a two-photon decay width for the a 2 (1320) of Γ/sub a 2(1320)→γγ/ = 1.17+-0. 15+-0.25 keV. 47 refs., 50 figs., 8 tabs

  16. Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy

    NARCIS (Netherlands)

    Jong, Imke G. de; Beilharz, Katrin; Kuipers, Oscar P.; Veening, Jan-Willem

    2011-01-01

    During the last few years scientists became increasingly aware that average data obtained from microbial population based experiments are not representative of the behavior, status or phenotype of single cells. Due to this new insight the number of single cell studies rises continuously. However,

  17. Time-Lapse, in Situ Imaging of Ice Crystal Growth Using Confocal Microscopy.

    Science.gov (United States)

    Marcellini, Moreno; Noirjean, Cecile; Dedovets, Dmytro; Maria, Juliette; Deville, Sylvain

    2016-11-30

    Ice crystals nucleate and grow when a water solution is cooled below its freezing point. The growth velocities and morphologies of the ice crystals depend on many parameters, such as the temperature of ice growth, the melting temperature, and the interactions of solutes with the growing crystals. Three types of morphologies may appear: dendritic, cellular (or fingerlike), or the faceted equilibrium form. Understanding and controlling which type of morphology is formed is essential in several domains, from biology to geophysics and materials science. Obtaining, in situ, three dimensional observations without introducing artifacts due to the experimental technique is nevertheless challenging. Here we show how we can use laser scanning confocal microscopy to follow in real-time the growth of smoothed and faceted ice crystals in zirconium acetate solutions. Both qualitative and quantitative observations can be made. In particular, we can precisely measure the lateral growth velocity of the crystals, a measure otherwise difficult to obtain. Such observations should help us understand the influence of the parameters that control the growth of ice crystals in various systems.

  18. Time-lapse monitoring of soil water content using electromagnetic conductivity imaging

    Science.gov (United States)

    The volumetric soil water content (VWC) is fundamental to agriculture. Unfortunately, the universally accepted thermogravimetric method is labour intensive and time-consuming to use for field-scale monitoring. Electromagnetic (EM) induction instruments have proven to be useful in mapping the spatio-...

  19. On the kinematics of the two-photon Cherenkov effect

    International Nuclear Information System (INIS)

    Afanas'ev, G.N.; Stepanovskij, Yu.P.

    2003-01-01

    We study the kinematics of the two-photon Cherenkov effect. In the general case, the emission angles of two photons satisfy certain inequalities and the corresponding radiation intensities are rather diffused. In special cases, when the above inequalities reduce to equalities, the emission angles of two photons are fixed and the corresponding radiation intensities should have sharp maxima at these angles. This makes easier the experimental study of the two-photon Cherenkov effect

  20. Time lapse photography as an approach to understanding glide avalanche activity

    Science.gov (United States)

    Hendrikx, Jordy; Peitzsch, Erich H.; Fagre, Daniel B.

    2012-01-01

    Avalanches resulting from glide cracks are notoriously difficult to forecast, but are a recurring problem for numerous avalanche forecasting programs. In some cases glide cracks are observed to open and then melt away in situ. In other cases, they open and then fail catastrophically as large, full-depth avalanches. Our understanding and management of these phenomena are currently limited. It is thought that an increase in the rate of snow gliding occurs prior to full-depth avalanche activity so frequent observation of glide crack movement can provide an index of instability. During spring 2011 in Glacier National Park, Montana, USA, we began an approach to track glide crack avalanche activity using a time-lapse camera focused on a southwest facing glide crack. This crack melted in-situ without failing as a glide avalanche, while other nearby glide cracks on north through southeast aspects failed. In spring 2012, a camera was aimed at a large and productive glide crack adjacent to the Going to the Sun Road. We captured three unique glide events in the field of view. Unfortunately, all of them either failed very quickly, or during periods of obscured view, so measurements of glide rate could not be obtained. However, we compared the hourly meteorological variables during the period of glide activity to the same variables prior to glide activity. The variables air temperature, relative humidity, air pressure, incoming and reflected long wave radiation, SWE, total precipitation, and snow depth were found to be statistically different for our cases examined. We propose that these are some of the potential precursors for glide avalanche activity, but do urge caution in their use, due to the simple approach and small data set size. It is hoped that by introducing a workable method to easily record glide crack movement, combined with ongoing analysis of the associated meteorological data, we will improve our understanding of when, or if, glide avalanche activity will ensue.

  1. ENVIRONMENTAL MONITORING OF LEAKS USING TIME LAPSED LONG ELECTRODE ELECTRICAL RESISTIVITY

    International Nuclear Information System (INIS)

    Myers, D.A.; Rucker, D.F.; Fink, J.B.; Loke, M.H.

    2009-01-01

    Highly industrialized areas pose challenges for surface electrical resistivity characterization due to metallic infrastructure. The infrastructure is typically more conductive than the desired targets and will mask the deeper subsurface information. These challenges may be minimized if steel-cased wells are used as long electrodes in the area near the target. We demonstrate a method of using long electrodes to electrically monitor a simulated leak from an underground storage tank with both synthetic examples and a field demonstration. The synthetic examples place a simple target of varying electrical properties beneath a very low resistivity layer. The layer is meant to replicate the effects of infrastructure. Both surface and long electrodes are tested on the synthetic domain. The leak demonstration for the field experiment is simulated by injecting a high conductivity fluid in a perforated well within the S tank farm at Hanford, and the resistivity measurements are made before and after the leak test. All data are processed in four dimensions, where a regularization procedure is applied in both the time and space domains. The synthetic test case shows that the long electrode ERM could detect relative changes in resistivity that are commensurate with the differing target properties. The surface electrodes, on the other hand, had a more difficult time matching the original target's footprint. The field results shows a lowered resistivity feature develop south of the injection site after cessation of the injections. The time lapsed regularization parameter has a strong influence on the differences in inverted resistivity between the pre and post injection datasets, but the interpretation of the target is consistent across all values of the parameter. The long electrode ERM method may provide a tool for near real-time monitoring of leaking underground storage tanks.

  2. Interpretaion of synthetic seismic time-lapse monitoring data for Korea CCS project based on the acoustic-elastic coupled inversion

    Science.gov (United States)

    Oh, J.; Min, D.; Kim, W.; Huh, C.; Kang, S.

    2012-12-01

    Recently, the CCS (Carbon Capture and Storage) is one of the promising methods to reduce the CO2 emission. To evaluate the success of the CCS project, various geophysical monitoring techniques have been applied. Among them, the time-lapse seismic monitoring is one of the effective methods to investigate the migration of CO2 plume. To monitor the injected CO2 plume accurately, it is needed to interpret seismic monitoring data using not only the imaging technique but also the full waveform inversion, because subsurface material properties can be estimated through the inversion. However, previous works for interpreting seismic monitoring data are mainly based on the imaging technique. In this study, we perform the frequency-domain full waveform inversion for synthetic data obtained by the acoustic-elastic coupled modeling for the geological model made after Ulleung Basin, which is one of the CO2 storage prospects in Korea. We suppose the injection layer is located in fault-related anticlines in the Dolgorae Deformed Belt and, for more realistic situation, we contaminate the synthetic monitoring data with random noise and outliers. We perform the time-lapse full waveform inversion in two scenarios. One scenario is that the injected CO2 plume migrates within the injection layer and is stably captured. The other scenario is that the injected CO2 plume leaks through the weak part of the cap rock. Using the inverted P- and S-wave velocities and Poisson's ratio, we were able to detect the migration of the injected CO2 plume. Acknowledgment This work was financially supported by the Brain Korea 21 project of Energy Systems Engineering, the "Development of Technology for CO2 Marine Geological Storage" program funded by the Ministry of Land, Transport and Maritime Affairs (MLTM) of Korea and the Korea CCS R&D Center (KCRC) grant funded by the Korea government (Ministry of Education, Science and Technology) (No. 2012-0008926).

  3. Two-photon excited UV fluorescence for protein crystal detection

    International Nuclear Information System (INIS)

    Madden, Jeremy T.; DeWalt, Emma L.; Simpson, Garth J.

    2011-01-01

    Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC

  4. Two-photon interference : spatial aspects of two-photon entanglement, diffraction, and scattering

    NARCIS (Netherlands)

    Peeters, Wouter Herman

    2010-01-01

    This dissertation contains scientific research within the realm of quantum optics, which is a branch of physics. An experimental and theoretical study is made of two-photon interference phenomena in various optical systems. Spatially entangled photon pairs are produced via the nonlinear optical

  5. A spirobifluorene-based two-photon fluorescence probe for mercury ions and its applications in living cells

    Energy Technology Data Exchange (ETDEWEB)

    Xiao, Haibo, E-mail: xiaohb@shnu.edu.cn; Zhang, Yanzhen; Zhang, Wu; Li, Shaozhi; Tan, Jingjing; Han, Zhongying

    2017-05-01

    A novel spirobifluorene derivative SPF-TMS, which containing dithioacetal groups and triphenylamine units, was synthesized. The probing behaviors toward various metal ions were investigated via UV/Vis absorption spectra as well as one-photon fluorescence changes. The results indicated that SPF-TMS exhibits high sensitivity and selectivity for mercury ions. The detection limit was at least 8.6 × 10{sup −8}M, which is excellent comparing with other optical sensors for Hg{sup 2+}. When measured by two-photon excited fluorescence technique in THF at 800 nm, the two-photon cross-section of SPF-TMS is 272 GM. Especially, upon reaction with mercury species, SPF-TMS yielded another two-photon dye SPF-DA. Both SPF-TMS and SPF-DA emit strong two-photon induced fluorescence and can be applied in cell imaging by two-photon microscopy. - Highlights: • We report a spirobifluorene-based molecule as two-photon fluorescent probe with large two-photon cross-section. • The molecule has exclusive selectivity and sensitivity for mercury species. • The molecule has large two-photon emission changes before and after addition of Hg{sup 2+}. • Both the probe and the mercury ion-promoted reaction product can be applied in cell imaging by two-photon microscopy.

  6. How to connect time-lapse recorded trajectories of motile microorganisms with dynamical models in continuous time

    DEFF Research Database (Denmark)

    Pedersen, Jonas Nyvold; Li, Liang; Gradinaru, Cristian

    2016-01-01

    We provide a tool for data-driven modeling of motility, data being time-lapse recorded trajectories. Several mathematical properties of a model to be found can be gleaned from appropriate model-independent experimental statistics, if one understands how such statistics are distorted by the finite...... of these effects that are valid for any reasonable model for persistent random motion. Our findings are illustrated with experimental data and Monte Carlo simulations....

  7. Modeling of time-lapse multi-scale seismic monitoring of CO2 injected into a fault zone to enhance the characterization of permeability in enhanced geothermal systems

    Science.gov (United States)

    Zhang, R.; Borgia, A.; Daley, T. M.; Oldenburg, C. M.; Jung, Y.; Lee, K. J.; Doughty, C.; Altundas, B.; Chugunov, N.; Ramakrishnan, T. S.

    2017-12-01

    Subsurface permeable faults and fracture networks play a critical role for enhanced geothermal systems (EGS) by providing conduits for fluid flow. Characterization of the permeable flow paths before and after stimulation is necessary to evaluate and optimize energy extraction. To provide insight into the feasibility of using CO2 as a contrast agent to enhance fault characterization by seismic methods, we model seismic monitoring of supercritical CO2 (scCO2) injected into a fault. During the CO2 injection, the original brine is replaced by scCO2, which leads to variations in geophysical properties of the formation. To explore the technical feasibility of the approach, we present modeling results for different time-lapse seismic methods including surface seismic, vertical seismic profiling (VSP), and a cross-well survey. We simulate the injection and production of CO2 into a normal fault in a system based on the Brady's geothermal field and model pressure and saturation variations in the fault zone using TOUGH2-ECO2N. The simulation results provide changing fluid properties during the injection, such as saturation and salinity changes, which allow us to estimate corresponding changes in seismic properties of the fault and the formation. We model the response of the system to active seismic monitoring in time-lapse mode using an anisotropic finite difference method with modifications for fracture compliance. Results to date show that even narrow fault and fracture zones filled with CO2 can be better detected using the VSP and cross-well survey geometry, while it would be difficult to image the CO2 plume by using surface seismic methods.

  8. Prediction of in-vitro developmental competence of early cleavage-stage mouse embryos with compact time-lapse equipment.

    Science.gov (United States)

    Pribenszky, Csaba; Losonczi, Eszter; Molnár, Miklós; Lang, Zsolt; Mátyás, Szabolcs; Rajczy, Klára; Molnár, Katalin; Kovács, Péter; Nagy, Péter; Conceicao, Jason; Vajta, Gábor

    2010-03-01

    Single blastocyst transfer is regarded as an efficient way to achieve high pregnancy rates and to avoid multiple pregnancies. Risk of cancellation of transfer due to a lack of available embryos may be reduced by early prediction of blastocyst development. Time-lapse investigation of mouse embryos shows that the time of the first and second cleavage (to the 2- and 3-cell stages, respectively) has a strong predictive value for further development in vitro, while cleavage from the 3-cell to the 4-cell stage has no predictive value. In humans, embryo fragmentation during preimplantation development has been associated with lower pregnancy rates and a higher incidence of developmental abnormalities. Analysis of time-lapse records shows that most fragmentation is reversible in the mouse and is resorbed in an average of 9 h. Daily or bi-daily microscopic checks of embryo development, applied routinely in human IVF laboratories, would fail to detect 36 or 72% of these fragmentations, respectively. Fragmentation occurring in a defined time frame has a strong predictive value for in-vitro embryo development. The practical compact system used in the present trial, based on the 'one camera per patient' principle, has eliminated the usual disadvantages of time-lapse investigations and is applicable for the routine follow-up of in-vitro embryo development. Copyright 2009 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  9. A time-series method for automated measurement of changes in mitotic and interphase duration from time-lapse movies.

    Directory of Open Access Journals (Sweden)

    Frederic D Sigoillot

    Full Text Available Automated time-lapse microscopy can visualize proliferation of large numbers of individual cells, enabling accurate measurement of the frequency of cell division and the duration of interphase and mitosis. However, extraction of quantitative information by manual inspection of time-lapse movies is too time-consuming to be useful for analysis of large experiments.Here we present an automated time-series approach that can measure changes in the duration of mitosis and interphase in individual cells expressing fluorescent histone 2B. The approach requires analysis of only 2 features, nuclear area and average intensity. Compared to supervised learning approaches, this method reduces processing time and does not require generation of training data sets. We demonstrate that this method is as sensitive as manual analysis in identifying small changes in interphase or mitotic duration induced by drug or siRNA treatment.This approach should facilitate automated analysis of high-throughput time-lapse data sets to identify small molecules or gene products that influence timing of cell division.

  10. Two photon processes in surface photovoltage spectroscopy

    International Nuclear Information System (INIS)

    Craig, R.P.; Thurgate, S.M.

    1996-01-01

    Full text: A significant mid-gap effect has been found in Surface Photovoltage Spectroscopy measurements of cleaved GaAs, InP and Si wafer which is normally interpreted as arising from transitions between surface states and band edges. This large mid-gap effect common to various materials is puzzling as such a high proportion of mid-gap states seems unlikely. Most theories of surface states predict states that tail from the band edges into the gap or states that have a well defined energy in the gap. None propose a large state exactly at E G /2. We recently investigated the variation in SPS spectra with flux density. We find a non-linear correlation in the magnitude of Band-Bending arising from mid-gap and band-gap photon energies. We suggest that the mid-gap feature is due to a two photon absorption process leading to carrier pair generation mediated by mid-gap states in the continuum of band-gap surface states

  11. STUDY OF TIME LAPSE IN FOREIGN BODY ASPIRATION IN RELATION TO CHEST X - RAY AND TYPE OF FOREIGN BODY

    Directory of Open Access Journals (Sweden)

    Salma

    2015-10-01

    Full Text Available INTRODUCTI ON: Foreign body aspiration in pediatrics is a potentially fatal accident which will continue until children explore their surroundings with their hand and mouth. Pediatric aspirations will persist until mankind exists. Not all foreign body aspirations are witnessed hence chances of delay in diagnosing an aspiration are high. Delay in diagnosis depends on site and character of foreign body aspirated. The chest x - ray findings and type of foreign body extracted vary depending on the duration the foreign body remains in airway . OBJECTIVE: To study the X - ray finding in pediatric airway aspiration and its relation to time lapse, the type and site of lodgment of foreign body extracted via bronchoscopy. The type of foreign body in relation to time lapse in aspiration. MATERIAL AND METHODS: This was a prospective study done in Bapuji child health institute and research center, JJM Medical College, Davangere . History and pre bronchoscopy x - Ray finding were noted for 65 children who were posted for suspicious bronchoscopy from August 2011 to September 2013. 11 children were excluded from study as they showed no foreign body on bronchoscopy. Time lapse in aspir ation and seeking medical care was noted. The bronchoscopic findings regarding site of foreign body lodgment and type of foreign body were recorded. The type of foreign body and variation of x - ray picture in relation to time lapse in aspiration were noted. Data collected was analyzed using descriptive statistics. RESULT: It was observed that mean age was 28 months. About 80% of the cases were between 1 to 3 years age. 82% (n=53/54 were radio lucent foreign body, only 1.5% (n=1/54 were radio o paque. Site of lodgment of foreign body was right main bronchus in 48% (n=26/54, left main bronchus 46% (n=25/54 , tracheal 1.85% (n=1/54, subglottic 1.85% (n=1/54, carinal 1.85% (n=1/54, multiple site i.e. left bronchus +right bronchus+ carinal 1.85% (n=1/54. Groundnut was most common

  12. Enhanced two-photon emission from a dressed biexciton

    International Nuclear Information System (INIS)

    Sánchez Muñoz, Carlos; Laussy, Fabrice P; Tejedor, Carlos; Valle, Elena del

    2015-01-01

    Radiative two-photon cascades from biexcitons in semiconductor quantum dots under resonant two-photon excitation are promising candidates for the generation of photon pairs. In this work, we propose a scheme to obtain two-photon emission that allows us to operate under very intense driving fields. This approach relies on the Purcell enhancement of two-photon virtual transitions between states of the biexciton dressed by the laser. The richness provided by the biexcitonic level structure allows to reach a variety of regimes, from antibunched and bunched photon pairs with polarization orthogonal to the driving field, to polarization entangled two-photon emission. This provides evidence that the general paradigm of two-photon emission from a ladder of dressed states can find interesting, particular implementations in a variety of systems. (paper)

  13. Development of a two photon microscope for tracking Drosophila larvae

    Science.gov (United States)

    Karagyozov, Doycho; Mihovilovic Skanata, Mirna; Gershow, Marc

    Current in vivo methods for measuring neural activity in Drosophila larva require immobilization of the animal. Although we can record neural signals while stimulating the sensory organs, we cannot read the behavioral output because we have prevented the animal from moving. Many research questions cannot be answered without observation of neural activity in behaving (freely-moving) animals. We incorporated a Tunable Acoustic Gradient (TAG) lens into a two-photon microscope to achieve a 70kHz axial scan rate, enabling volumetric imaging at tens of hertz. We then implemented a tracking algorithm based on a Kalman filter to maintain the neurons of interest in the field of view and in focus during the rapid three dimensional motion of a free larva. Preliminary results show successful tracking of a neuron moving at speeds reaching 500 μm/s. NIH Grant 1DP2EB022359 and NSF Grant PHY-1455015.

  14. Clinical multiphoton tomography and clinical two-photon microendoscopy

    Science.gov (United States)

    König, Karsten; Bückle, Rainer; Weinigel, Martin; Elsner, Peter; Kaatz, Martin

    2009-02-01

    We report on applications of high-resolution clinical multiphoton tomography based on the femtosecond laser system DermaInspectTM with its flexible mirror arm in Australia, Asia, and Europe. Applications include early detection of melanoma, in situ tracing of pharmacological and cosmetical compounds including ZnO nanoparticles in the epidermis and upper dermis, the determination of the skin aging index SAAID as well as the study of the effects of anti-aging products. In addition, first clinical studies with novel rigid high-NA two-photon 1.6 mm GRIN microendoscopes have been conducted to study the effect of wound healing in chronic wounds (ulcus ulcera) as well as to perform intrabody imaging with subcellular resolution in small animals.

  15. Molecular engineering of two-photon fluorescent probes for bioimaging applications

    Science.gov (United States)

    Liu, Hong-Wen; Liu, Yongchao; Wang, Peng; Zhang, Xiao-Bing

    2017-03-01

    During the past two decades, two-photon microscopy (TPM), which utilizes two near-infrared photons as the excitation source, has emerged as a novel, attractive imaging tool for biological research. Compared with one-photon microscopy, TPM offers several advantages, such as lowering background fluorescence in living cells and tissues, reducing photodamage to biosamples, and a photobleaching phenomenon, offering better 3D spatial localization, and increasing penetration depth. Small-molecule-based two-photon fluorescent probes have been well developed for the detection and imaging of various analytes in biological systems. In this review, we will give a general introduction of molecular engineering of two-photon fluorescent probes based on different fluorescence response mechanisms for bioimaging applications during the past decade. Inspired by the desired advantages of small-molecule two-photon fluorescent probes in biological imaging applications, we expect that more attention will be devoted to the development of new two-photon fluorophores and applications of TPM in areas of bioanalysis and disease diagnosis.

  16. Two-photon decay in heavy atoms and ions

    International Nuclear Information System (INIS)

    Mokler, P.H.; Dunford, R.W

    2003-08-01

    We review the status of and comment on current developments in the field of two-photon decay in atomic physics research. Recent work has focused on two-photon decays in highly-charged ions and two-photon decay of inner-shell vacancies in heavy neutral atoms. We emphasize the importance of measuring the shape of the continuum emission in two-photon decay as a probe of relativistic effects in the strong central fields found in heavy atomic systems. New experimental approaches and their consequences will be discussed. (orig.)

  17. Estimation of soil hydraulic parameters in the field by integrated hydrogeophysical inversion of time-lapse ground-penetrating radar data

    KAUST Repository

    Jadoon, Khan; Weihermü ller, Lutz; Scharnagl, Benedikt; Kowalsky, Michael B.; Bechtold, Michel; Hubbard, Susan S.; Vereecken, Harry; Lambot, Sé bastien

    2012-01-01

    An integrated hydrogeophysical inversion approach was used to remotely infer the unsaturated soil hydraulic parameters from time-lapse ground-penetrating radar (GPR) data collected at a fixed location over a bare agricultural field. The GPR model

  18. Resonance production in two-photon interactions

    International Nuclear Information System (INIS)

    Roe, N.A.

    1989-02-01

    Resonance production in two-photon interactions is studied using data collected with the ASP detector at the PEP e + e/sup /minus// storage ring located at the Stanford Linear Accelerator Center. The ASP detector is a non-magnetic lead-glass calorimeter constructed from 632 lead-glass bars. It covers 94% of 4π in solid angle, extending to within 20/degree/ of the beamline. Lead-scintillator calorimeters extend the coverage to within 21 mr of the beamline on both sides. Energy resolution of √E/10%, where E is the energy is GeV, is achieved for electrons and photons in the lead-glass calorimeter, and particle trajectories are reconstructed with high efficiency. A total luminosity of 108 pb/sup /minus/1/ was collected with the ASP detector at a center-of-mass energy of 29 GeV. The observed process is e + e/sup /minus// → e + e/sup /minus//γ*γ* → e + e/sup /minus//X, is a pseudoscalar resonance (J/sup PC/ = 0/sup /minus/+/) and γ* is a virtual (mass /ne/ 0) photon. The outgoing electrons scatter down the beampipe and are not detected. The observed resonances are the /eta/ and /eta/' mesons, with masses of 549 and 958 MeV, respectively. They are detected in the γγ decay mode; a total of 2380 +- 49 /eta/ → γγ and 568 +- 26 /eta/' → γγ events are observed. From the number of events, the detection efficiency, and the calculated production cross sections the radiative widths, Γ/sub γγ/, of the /eta/ and /eta/' were measured and found to be: Γ/sub γγ/(/eta/) = .481 +- .010 +- .047keV and Γ/sub γγ/(/eta/') = 4.71 +- .22 +- .70keV. These results are in good agreement with the world average values. 67 refs., 42 figs., 20 tabs

  19. Resonance production in two-photon interactions

    Energy Technology Data Exchange (ETDEWEB)

    Roe, N.A.

    1989-02-01

    Resonance production in two-photon interactions is studied using data collected with the ASP detector at the PEP e/sup +/e/sup /minus// storage ring located at the Stanford Linear Accelerator Center. The ASP detector is a non-magnetic lead-glass calorimeter constructed from 632 lead-glass bars. It covers 94% of 4..pi.. in solid angle, extending to within 20/degree/ of the beamline. Lead-scintillator calorimeters extend the coverage to within 21 mr of the beamline on both sides. Energy resolution of ..sqrt..E/10%, where E is the energy is GeV, is achieved for electrons and photons in the lead-glass calorimeter, and particle trajectories are reconstructed with high efficiency. A total luminosity of 108 pb/sup /minus/1/ was collected with the ASP detector at a center-of-mass energy of 29 GeV. The observed process is e/sup +/e/sup /minus// ..-->.. e/sup +/e/sup /minus//..gamma..*..gamma..* ..-->.. e/sup +/e/sup /minus//X, is a pseudoscalar resonance (J/sup PC/ = 0/sup /minus/+/) and ..gamma..* is a virtual (mass /ne/ 0) photon. The outgoing electrons scatter down the beampipe and are not detected. The observed resonances are the /eta/ and /eta/' mesons, with masses of 549 and 958 MeV, respectively. They are detected in the ..gamma gamma.. decay mode; a total of 2380 +- 49 /eta/ ..-->.. ..gamma gamma.. and 568 +- 26 /eta/' ..-->.. ..gamma gamma.. events are observed. From the number of events, the detection efficiency, and the calculated production cross sections the radiative widths, GAMMA/sub ..gamma gamma../, of the /eta/ and /eta/' were measured and found to be: GAMMA/sub ..gamma gamma../(/eta/) = .481 +- .010 +- .047keV and GAMMA/sub ..gamma gamma../(/eta/') = 4.71 +- .22 +- .70keV. These results are in good agreement with the world average values. 67 refs., 42 figs., 20 tabs.

  20. Effect of time lapse on the diagnostic accuracy of cone beam computed tomography for detection of vertical root fractures

    Energy Technology Data Exchange (ETDEWEB)

    Eskandarloo, Amir; Shokri, Abbas, E-mail: Dr.a.shokri@gmail.com [Dental Research Center, Department of Oral and Maxillofacial Radiology, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Asl, Amin Mahdavi [Department of Oral and Maxillofacial Radiology, Golestan University of Medical Sciences, Gorgan (Iran, Islamic Republic of); Jalalzadeh, Mohsen [Department of Endodontics, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Tayari, Maryam [Department of Pedodontics, Golestan University of Medical Sciences, Gorgan (Iran, Islamic Republic of); Hosseinipanah, Mohammad [Department of Anatomy, School of Medicine, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of); Fardmal, Javad [Research Center for Health Sciences and Department of Epidemiology and Biostatistics, School of Public Health, Hamadan University of Medical Sciences, Hamadan (Iran, Islamic Republic of)

    2016-01-15

    Accurate and early diagnosis of vertical root fractures (VRFs) is imperative to prevent extensive bone loss and unnecessary endodontic and prosthodontic treatments. The aim of this study was to assess the effect of time lapse on the diagnostic accuracy of cone beam computed tomography (CBCT) for VRFs in endodontically treated dog’s teeth. Forty eight incisors and premolars of three adult male dogs underwent root canal therapy. The teeth were assigned to two groups: VRFs were artificially induced in the first group (n=24) while the teeth in the second group remained intact (n=24). The CBCT scans were obtained by NewTom 3G unit immediately after inducing VRFs and after one, two, three, four, eight, 12 and 16 weeks. Three oral and maxillofacial radiologists blinded to the date of radiographs assessed the presence/absence of VRFs on CBCT scans. The sensitivity, specificity and accuracy values were calculated and data were analyzed using SPSS v.16 software and ANOVA. The total accuracy of detection of VRFs immediately after surgery, one, two, three, four, eight, 12 and 16 weeks was 67.3%, 68.7%, 66.6%, 64.6%, 64.5%, 69.4%, 68.7%, 68% respectively. The effect of time lapse on detection of VRFs was not significant (p>0.05). Overall sensitivity, specificity and accuracy of CBCT for detection of VRFs were 74.3%, 62.2%, 67.2% respectively. Cone beam computed tomography is a valuable tool for detection of VRFs. Time lapse (four months) had no effect on detection of VRFs on CBCT scans. (author)

  1. Effect of time lapse on the diagnostic accuracy of cone beam computed tomography for detection of vertical root fractures

    International Nuclear Information System (INIS)

    Eskandarloo, Amir; Shokri, Abbas; Asl, Amin Mahdavi; Jalalzadeh, Mohsen; Tayari, Maryam; Hosseinipanah, Mohammad; Fardmal, Javad

    2016-01-01

    Accurate and early diagnosis of vertical root fractures (VRFs) is imperative to prevent extensive bone loss and unnecessary endodontic and prosthodontic treatments. The aim of this study was to assess the effect of time lapse on the diagnostic accuracy of cone beam computed tomography (CBCT) for VRFs in endodontically treated dog’s teeth. Forty eight incisors and premolars of three adult male dogs underwent root canal therapy. The teeth were assigned to two groups: VRFs were artificially induced in the first group (n=24) while the teeth in the second group remained intact (n=24). The CBCT scans were obtained by NewTom 3G unit immediately after inducing VRFs and after one, two, three, four, eight, 12 and 16 weeks. Three oral and maxillofacial radiologists blinded to the date of radiographs assessed the presence/absence of VRFs on CBCT scans. The sensitivity, specificity and accuracy values were calculated and data were analyzed using SPSS v.16 software and ANOVA. The total accuracy of detection of VRFs immediately after surgery, one, two, three, four, eight, 12 and 16 weeks was 67.3%, 68.7%, 66.6%, 64.6%, 64.5%, 69.4%, 68.7%, 68% respectively. The effect of time lapse on detection of VRFs was not significant (p>0.05). Overall sensitivity, specificity and accuracy of CBCT for detection of VRFs were 74.3%, 62.2%, 67.2% respectively. Cone beam computed tomography is a valuable tool for detection of VRFs. Time lapse (four months) had no effect on detection of VRFs on CBCT scans. (author)

  2. A multimethod Global Sensitivity Analysis to aid the calibration of geomechanical models via time-lapse seismic data

    Science.gov (United States)

    Price, D. C.; Angus, D. A.; Garcia, A.; Fisher, Q. J.; Parsons, S.; Kato, J.

    2018-03-01

    Time-lapse seismic attributes are used extensively in the history matching of production simulator models. However, although proven to contain information regarding production induced stress change, it is typically only loosely (i.e. qualitatively) used to calibrate geomechanical models. In this study we conduct a multimethod Global Sensitivity Analysis (GSA) to assess the feasibility and aid the quantitative calibration of geomechanical models via near-offset time-lapse seismic data. Specifically, the calibration of mechanical properties of the overburden. Via the GSA, we analyse the near-offset overburden seismic traveltimes from over 4000 perturbations of a Finite Element (FE) geomechanical model of a typical High Pressure High Temperature (HPHT) reservoir in the North Sea. We find that, out of an initially large set of material properties, the near-offset overburden traveltimes are primarily affected by Young's modulus and the effective stress (i.e. Biot) coefficient. The unexpected significance of the Biot coefficient highlights the importance of modelling fluid flow and pore pressure outside of the reservoir. The FE model is complex and highly nonlinear. Multiple combinations of model parameters can yield equally possible model realizations. Consequently, numerical calibration via a large number of random model perturbations is unfeasible. However, the significant differences in traveltime results suggest that more sophisticated calibration methods could potentially be feasible for finding numerous suitable solutions. The results of the time-varying GSA demonstrate how acquiring multiple vintages of time-lapse seismic data can be advantageous. However, they also suggest that significant overburden near-offset seismic time-shifts, useful for model calibration, may take up to 3 yrs after the start of production to manifest. Due to the nonlinearity of the model behaviour, similar uncertainty in the reservoir mechanical properties appears to influence overburden

  3. The Potential Impact of Biofield Treatment on Human Brain Tumor Cells: A Time-Lapse Video Microscopy

    OpenAIRE

    Trivedi, Mahendra

    2015-01-01

    Study background: Glioblastoma (GBM) is the most common subtype of primary brain tumor in adults. The aim was to evaluate the impact of biofield treatment potential on human GBM and non-GBM brain cells using two time-lapse video microscopy technique. Methods: The human brain tumor, GBM cultured cells were divided into two groups viz. GBM control and GBM treatment. Similarly, human normal brain cultured cells (non-GBM) were taken and divided into two groups viz. non- GBM control and non-GB...

  4. A Dynamic Programming Model for Optimizing Frequency of Time-Lapse Seismic Monitoring in Geological CO2 Storage

    Science.gov (United States)

    Bhattacharjya, D.; Mukerji, T.; Mascarenhas, O.; Weyant, J.

    2005-12-01

    Designing a cost-effective and reliable monitoring program is crucial to the success of any geological CO2 storage project. Effective design entails determining both, the optimal measurement modality, as well as the frequency of monitoring the site. Time-lapse seismic provides the best spatial coverage and resolution for reservoir monitoring. Initial results from Sleipner (Norway) have demonstrated effective monitoring of CO2 plume movement. However, time-lapse seismic is an expensive monitoring technique especially over the long term life of a storage project and should be used judiciously. We present a mathematical model based on dynamic programming that can be used to estimate site-specific optimal frequency of time-lapse surveys. The dynamics of the CO2 sequestration process are simplified and modeled as a four state Markov process with transition probabilities. The states are M: injected CO2 safely migrating within the target zone; L: leakage from the target zone to the adjacent geosphere; R: safe migration after recovery from leakage state; and S: seepage from geosphere to the biosphere. The states are observed only when a monitoring survey is performed. We assume that the system may go to state S only from state L. We also assume that once observed to be in state L, remedial measures are always taken to bring it back to state R. Remediation benefits are captured by calculating the expected penalty if CO2 seeped into the biosphere. There is a trade-off between the conflicting objectives of minimum discounted costs of performing the next time-lapse survey and minimum risk of seepage and its associated costly consequences. A survey performed earlier would spot the leakage earlier. Remediation methods would have been utilized earlier, resulting in savings in costs attributed to excessive seepage. On the other hand, there are also costs for the survey and remedial measures. The problem is solved numerically using Bellman's optimality principal of dynamic

  5. Molecular photosensitisers for two-photon photodynamic therapy.

    Science.gov (United States)

    Bolze, F; Jenni, S; Sour, A; Heitz, V

    2017-11-30

    Two-photon excitation has attracted the attention of biologists, especially after the development of two-photon excited microscopy in the nineties. Since then, new applications have rapidly emerged such as the release of biologically active molecules and photodynamic therapy (PDT) using two-photon excitation. PDT, which requires a light-activated drug (photosensitiser), is a clinically approved and minimally invasive treatment for cancer and for non-malignant diseases. This feature article focuses on the engineering of molecular two-photon photosensitisers for PDT, which should bring important benefits to the treatment, increase the treatment penetration depth with near-infrared light excitation, improve the spatial selectivity and reduce the photodamage to healthy tissues. After an overview of the two-photon absorption phenomenon and the methods to evaluate two-photon induced phototoxicity on cell cultures, the different classes of photosensitisers described in the literature are discussed. The two-photon PDT performed with historical one-photon sensitisers are briefly presented, followed by specifically engineered cyclic tetrapyrrole photosensitisers, purely organic photosensitisers and transition metal complexes. Finally, targeted two-photon photosensitisers and theranostic agents that should enhance the selectivity and efficiency of the treatment are discussed.

  6. Two-photon stimulated emission and pulse amplification

    International Nuclear Information System (INIS)

    Yuen, H.P.

    1975-01-01

    Threshold conditions are given for the sustained operation of standing-wave and long-pulse traveling-wave two-photon lasers. Pulse shortening in long-pulse two-photon amplification, a behavior absent in the one-photon case, is also demonstrated analytically. (U.S.)

  7. Novel triphenylamine-cored two-photon absorbing dyes for labeling of biomolecules

    International Nuclear Information System (INIS)

    Xiao Haibo; Mei Chong; Wang Yaochuan; Li, Hui; Qian Shixiong; Yin Hongyao; Xu Zhisong

    2011-01-01

    Highlights: → Two novel triphenylamine-cored chromophores were synthesized. → These two dyes have sizable two-photon absorption cross-section at 800 nm. → They possess reasonable water solubility and are suitable as labels in aqueous biological environments. → These dyes have strong chelating ability. → They display a large set of reactivity for coupling to biomolecules. - Abstract: Two novel, V-shaped and Y-shaped dipicolinate derivatives branched from triphenylamine, {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinyl]}-N-phenyl-N-{4- [(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl)vinylphenyl]}aniline (1) and {4-[(E)-2-(2,6-dimethoxycarbonylpyridin-4-yl) vinyl]}-N,N-bis {4-[(E)-2-(2,6-dimethoxycarbonyl pyridin-4-yl)vinylphenyl]}aniline (2) were synthesized. These compounds were designed for large two-photon absorption and in particular for labeling of biomolecules. Their linear absorption, fluorescence properties and their two-photon absorption properties as well as two-photon fluorescence cell imaging were examined. When excited at 800 nm, the two-photon absorption cross-section values of chromophores 1 and 2 in THF were 208 GM, 376 GM, respectively. These two-photon absorbing dyes possess reasonable water solubility, strong chelating ability and display a large set of reactivity for coupling to biomolecules, which are apparently due to the two methoxycarbonyl groups in pyridine ring. This work suggests that chromophores 1 and 2 are promising labels potentially applicable for the tracking of biomolecules using two-photon scanning microscopy.

  8. Real-time intravital imaging of pH variation associated with osteoclast activity.

    Science.gov (United States)

    Maeda, Hiroki; Kowada, Toshiyuki; Kikuta, Junichi; Furuya, Masayuki; Shirazaki, Mai; Mizukami, Shin; Ishii, Masaru; Kikuchi, Kazuya

    2016-08-01

    Intravital imaging by two-photon excitation microscopy (TPEM) has been widely used to visualize cell functions. However, small molecular probes (SMPs), commonly used for cell imaging, cannot be simply applied to intravital imaging because of the challenge of delivering them into target tissues, as well as their undesirable physicochemical properties for TPEM imaging. Here, we designed and developed a functional SMP with an active-targeting moiety, higher photostability, and a fluorescence switch and then imaged target cell activity by injecting the SMP into living mice. The combination of the rationally designed SMP with a fluorescent protein as a reporter of cell localization enabled quantitation of osteoclast activity and time-lapse imaging of its in vivo function associated with changes in cell deformation and membrane fluctuations. Real-time imaging revealed heterogenic behaviors of osteoclasts in vivo and provided insights into the mechanism of bone resorption.

  9. The development of efficient two-photon singlet oxygen sensitizers

    DEFF Research Database (Denmark)

    Nielsen, Christian Benedikt

    The development of efficient two-photon singlet oxygen sensitizers is addressed focusing on organic synthesis. Photophysical measurements were carried out on new lipophilic molecules, where two-photon absorption cross sections and singlet oxygen quantumyields were measured. Design principles...... for making efficient two-photon singlet oxygen sensitizers were then constructed from these results. Charge-transfer in the excited state of the prepared molecules was shown to play a pivotal role in the generationof singlet oxygen. This was established through studies of substituent effects on both...... the singlet oxygen yield and the two-photon absorption cross section, where it was revealed that a careful balancing of the amount of charge transfer present in theexcited state of the sensitizer is necessary to obtain both a high singlet oxygen quantum yield and a high two-photon cross section. An increasing...

  10. Determination of residual oil saturation from time-lapse pulsed neutron capture logs in a large sandstone reservoir

    International Nuclear Information System (INIS)

    Syed, E.V.; Salaita, G.N.; McCaffery, F.G.

    1991-01-01

    Cased hole logging with pulsed neutron tools finds extensive use for identifying zones of water breakthrough and monitoring oil-water contacts in oil reservoirs being depleted by waterflooding or natural water drive. Results of such surveys then find direct use for planning recompletions and water shutoff treatments. Pulsed neutron capture (PNC) logs are useful for estimating water saturation changes behind casing in the presence of a constant, high-salinity environment. PNC log surveys run at different times, i.e., in a time-lapse mode, are particularly amenable to quantitative analysis. The combined use of the original open hole and PNC time-lapse log information can then provide information on remaining or residual oil saturations in a reservoir. This paper reports analyses of historical pulsed neutron capture log data to assess residual oil saturation in naturally water-swept zones for selected wells from a large sandstone reservoir in the Middle East. Quantitative determination of oil saturations was aided by PNC log information obtained from a series of tests conducted in a new well in the same field

  11. Estimation of soil hydraulic parameters by integrated hydrogeophysical inversion of time-lapse GPR data measured at Selhausen, Germany

    KAUST Repository

    Jadoon, Khan

    2012-06-01

    We present an integrated hydrogeophysical inversion approach that uses time-lapse off-ground ground-penetrating radar (GPR) data to estimate soil hydraulic parameters, and apply it to a dataset collected in the field. Off-ground GPR data are mainly sensitive to the near-surface water content profile and dynamics, and are thus related to soil hydraulic parameters, such as the parameters of the hydraulic conductivity and water retention functions. The hydrological simulator HYDRUS 1-D was used with a two-layer single- and dual-porosity model. To monitor the soil water content dynamics, time-lapse GPR and time domain reflectometry (TDR) measurements were performed, whereby only GPR data was used in the inversion. The dual porosity model provided better results compared to the single porosity model for describing the soil water dynamics, which is supported by field observations of macropores. Furthermore, the GPR-derived water content profiles reconstructed from the integrated hydrogeophysical inversion were in good agreement with TDR observations. These results suggest that the proposed method is promising for non-invasive characterization of the shallow subsurface hydraulic properties and monitoring water dynamics at the field scale.

  12. A comparison of the cytotoxic activity of eosinophils and other cells by 51chromium release and time lapse microcinematography

    International Nuclear Information System (INIS)

    Sanderson, C.J.; Thomas, J.A.

    1978-01-01

    Antibody dependent cytotoxicity of chicken erythrocytes by purified rat eosinophils, neutrophils, macrophages and K cells has been compared by 51 Cr release and time lapse microcinematography. Techniques have been developed for purifying these effector cell types. Both eosinophils and neutrophils caused rapid release of 51 Cr from erythrocytes. Time lapse observations indicated that this was the result of phagocytosis. Eosinophils showed rapid membrane movement and repeatedly engulfed and regurgitated the erythrocytes. On the other hand, neutrophils became quiescent after phagocytosing erythrocytes, and remained quiescent until the remains of the cell were expelled. Neutrophils presumably have a mechanism for the release of soluble material, as 51 Cr was released rapidly. Macrophages showed a similar quiescence after phagocytosis, but in these cells there was apparently no rapid mechanism to expel material, as there was no significant 51 Cr release over 20 h. K cells appeared to damage chicken erythrocytes more slowly than they destroyed tumour cells. Mast cells caused antibody-independent cytotoxicity which can be attributed to the release of toxic materials. None of these effector cells produced the type of lysis seen with antibody and complement. (author)

  13. Gigavision - A weatherproof, multibillion pixel resolution time-lapse camera system for recording and tracking phenology in every plant in a landscape

    Science.gov (United States)

    Brown, T.; Borevitz, J. O.; Zimmermann, C.

    2010-12-01

    retrieved at bi-weekly intervals. Our longer-term goal is to make gigapixel time-lapse datasets available online in an interactive interface that layers plant-level phenology data with gigapixel resolution images, genomic sequence data from individual plants with weather and other abitotic sensor data. Co-visualization of all of these data types provides researchers with a powerful new tool for examining complex ecological interactions across scales from the individual to the ecosystem. We will present detailed phenostage data from more than 100 plants of multiple species from our Gigavision timelapse camera at our “Big Blowout East” field site in the Indiana Dunes State Park, IN. This camera has been recording three to four 700 million pixel images a day since February 28, 2010. The camera field of view covers an area of about 7 hectares resulting in an average image resolution of about 1 pixel per centimeter over the entire site. We will also discuss some of the many technological challenges with developing and maintaining these types of hardware systems, collecting quantitative data from gigapixel resolution time-lapse data and effectively managing terabyte-sized datasets of millions of images.

  14. Insights into esophagus tissue architecture using two-photon confocal microscopy

    Science.gov (United States)

    Liu, Nenrong; Wang, Yue; Feng, Shangyuan; Chen, Rong

    2013-08-01

    In this paper, microstructures of human esophageal mucosa were evaluated using the two-photon laser scanning confocal microscopy (TPLSCM), based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). The distribution of epithelial cells, muscle fibers of muscularis mucosae has been distinctly obtained. Furthermore, esophageal submucosa characteristics with cancer cells invading into were detected. The variation of collagen, elastin and cancer cells is very relevant to the pathology in esophagus, especially early esophageal cancer. Our experimental results indicate that the MPM technique has the much more advantages for label-free imaging, and has the potential application in vivo in the clinical diagnosis and monitoring of early esophageal cancer.

  15. Circular dichroism probed by two-photon fluorescence microscopy in enantiopure chiral polyfluorene thin films

    NARCIS (Netherlands)

    Savoini, M.; Wu, Xiaofei; Celebrano, M.; Ziegler, J.; Biagioni, P.; Meskers, S.C.J.; Duo, L.; Hecht, B.; Finazzi, M.

    2012-01-01

    Two-photon fluorescence scanning confocal microscopy sensitive to circular dichroism with a diffraction-limited resolution well below 500 nm is demonstrated. With this method, the spatial variation of the circular dichroism of thermally annealed chiral polyfluorene thin films has been imaged. We

  16. Dependence of the two-photon photoluminescence yield of gold nanostructures on the laser pulse duration

    DEFF Research Database (Denmark)

    Biagioni, P.; Celebrano, M.; Savoini, M.

    2009-01-01

    Two-photon photoluminescence (TPPL) from gold nanostructures is becoming one of the most relevant tools for plasmon-assisted biological imaging and photothermal therapy as well as for the investigation of plasmonic devices. Here we study the yield of TPPL as a function of the temporal width δ of ...

  17. Two-photon luminescence microscopy of field enhancement at gold nanoparticles

    DEFF Research Database (Denmark)

    Beermann, Jonas; Bozhevolnyi, Sergey I.

    2005-01-01

    Using a reflection scanning optical microscope detecting two-photon luminescence (TPL) we have imaged square gold bumps positioned in a periodic array either on a smooth gold film or directly on a glass substrate. The second-harmonic (SH) and TPL response from these structures show both...

  18. Two-photon processes in highly charged ions

    International Nuclear Information System (INIS)

    Jahrsetz, Thorsten

    2015-01-01

    Two-photon processes are atomic processes in which an atom interacts simultaneously with two photons. Such processes describe a wide range of phenomena, such as two-photon decay and elastic or inelastic scattering of photons. In recent years two-photon processes involving highly charged heavy ions have become an active area of research. Such studies do not only consider the total transition or scattering rates but also their angular and polarization dependence. To support such examinations in this thesis I present a theoretical framework to describe these properties in all two-photon processes with bound initial and final states and involving heavy H-like or He-like ions. I demonstrate how this framework can be used in some detailed studies of different two-photon processes. Specifically a detailed analysis of two-photon decay of H-like and He-like ions in strong external electromagnetic fields shows the importance of considering the effect of such fields for the physics of such systems. Furthermore I studied the elastic Rayleigh as well as inelastic Raman scattering by heavy H-like ions. I found a number of previously unobserved phenomena in the angular and polarization dependence of the scattering cross-sections that do not only allow to study interesting details of the electronic structure of the ion but might also be useful for the measurement of weak physical effects in such systems.

  19. Two-photon processes in highly charged ions

    Energy Technology Data Exchange (ETDEWEB)

    Jahrsetz, Thorsten

    2015-03-05

    Two-photon processes are atomic processes in which an atom interacts simultaneously with two photons. Such processes describe a wide range of phenomena, such as two-photon decay and elastic or inelastic scattering of photons. In recent years two-photon processes involving highly charged heavy ions have become an active area of research. Such studies do not only consider the total transition or scattering rates but also their angular and polarization dependence. To support such examinations in this thesis I present a theoretical framework to describe these properties in all two-photon processes with bound initial and final states and involving heavy H-like or He-like ions. I demonstrate how this framework can be used in some detailed studies of different two-photon processes. Specifically a detailed analysis of two-photon decay of H-like and He-like ions in strong external electromagnetic fields shows the importance of considering the effect of such fields for the physics of such systems. Furthermore I studied the elastic Rayleigh as well as inelastic Raman scattering by heavy H-like ions. I found a number of previously unobserved phenomena in the angular and polarization dependence of the scattering cross-sections that do not only allow to study interesting details of the electronic structure of the ion but might also be useful for the measurement of weak physical effects in such systems.

  20. Exciton molecule in semiconductors by two-photon absorption

    International Nuclear Information System (INIS)

    Arya, K.; Hassan, A.R.

    1976-07-01

    Direct creation of bi-exciton states by two-photon absorption in direct gap semiconductors is investigated theoretically. A numerical application to the case of CuCl shows that the two-photon absorption coefficient for bi-excitonic transitions is larger than that for two-photon interband transitions by three orders of magnitude. It becomes comparable to that for one-photon excitonic transitions for available laser intensities. The main contribution to this enhancement of the absorption coefficient for the transitions to the bi-exciton states is found to be from the resonance effect

  1. High-resolution time-lapse tomography of rat vertebrae during compressive loading: deformation response analysis

    Czech Academy of Sciences Publication Activity Database

    Fíla, Tomáš; Kytýř, Daniel; Zlámal, Petr; Kumpová, Ivana; Doktor, Tomáš; Koudelka_ml., Petr; Jiroušek, Ondřej

    2014-01-01

    Roč. 9, May (2014), C05054 ISSN 1748-0221. [International workshop on radiation imaging detectors /15./. Paris, 23.06.2013-27.06.2013] R&D Projects: GA ČR(CZ) GAP105/10/2305; GA MŠk(CZ) LO1219 Institutional support: RVO:68378297 Keywords : image reconstruction in medical imaging * computerized tomography * computed radiography Subject RIV: JJ - Other Materials Impact factor: 1.399, year: 2014 http://iopscience.iop.org/1748-0221/9/05/C05054

  2. Phosphorescent probes for two-photon microscopy of oxygen (Conference Presentation)

    Science.gov (United States)

    Vinogradov, Sergei A.; Esipova, Tatiana V.

    2016-03-01

    The ability to quantify oxygen in vivo in 3D with high spatial and temporal resolution is much needed in many areas of biological research. Our laboratory has been developing the phosphorescence quenching technique for biological oximetry - an optical method that possesses intrinsic microscopic capability. In the past we have developed dendritically protected oxygen probes for quantitative imaging of oxygen in tissue. More recently we expanded our design on special two-photon enhanced phosphorescent probes. These molecules brought about first demonstrations of the two-photon phosphorescence lifetime microscopy (2PLM) of oxygen in vivo, providing new information for neouroscience and stem cell biology. However, current two-photon oxygen probes suffer from a number of limitations, such as sub-optimal brightness and high cost of synthesis, which dramatically reduce imaging performance and limit usability of the method. In this paper we discuss principles of 2PLM and address the interplay between the probe chemistry, photophysics and spatial and temporal imaging resolution. We then present a new approach to brightly phosphorescent chromophores with internally enhanced two-photon absorption cross-sections, which pave a way to a new generation of 2PLM probes.

  3. Highlights from the SoilCAM project: Soil Contamination, Advanced integrated characterisation and time-lapse Monitoring

    Science.gov (United States)

    French, H. K.; van der Zee, S. E. A. T. M.; Wehrer, M.; Godio, A.; Pedersen, L. B.; Toscano, G.

    2012-04-01

    The SoilCAM project (Soil Contamination, Advanced integrated characterisation and time-lapse Monitoring 2008-2012, EU-FP7-212663) is aimed at improving current methods for monitoring contaminant distribution and biodegradation in the subsurface. At two test sites, Oslo airport Gardermoen in Norway and the Trecate site in Italy, a number of geophysical techniques, lysimeter and other soil and water sampling techniques as well as numerical flow and transport modelling have been combined at different scales in order to characterise flow transport processes in the unsaturated and saturated zones. Laboratory experiments have provided data on physical and bio-geo-chemical parameters for use in models and to select remediation methods. The geophysical techniques were used to map geological heterogeneities and also conduct time-lapse measurements of processes in the unsaturated zone. Both cross borehole and surface electrodes were used for electrical resistivity and induced polarisation surveys. The geophysical surveys showed clear indications of areas highly affected by de-icing chemicals along the runway at Oslo airport. The time lapse measurements along the runway at the airport show infiltration patterns during snowmelt and are used to validate 2D unsaturated flow and transport simulations using SUTRA. The Orchestra model is used to describe the complex interaction between bio-geo-chemical processes in a 1D profile along the runway. The presence of installations such as a membrane along the runway highly affects the flow pattern and challenges the capacity of the numerical code. Smaller scale field site measurements have revealed the increase of iron and manganese during degradation of de-icing chemicals. The use of Nitrate to increase red-ox potential was tested, but results have not been analysed yet. So far it cannot be concluded that degradation process can be quantified indirectly by geophysical monitoring. At the Trecate site a combination of georadar, electrical

  4. Live Cells as Dynamic Laboratories: Time Lapse Raman Spectral Microscopy of Nanoparticles with Both IgE Targeting and pH-Sensing Functions

    Directory of Open Access Journals (Sweden)

    Kristy L. Nowak-Lovato

    2012-01-01

    Full Text Available This review captures the use of live cells as dynamic microlaboratories through implementation of labeled nanoparticles (nanosensors that have both sensing and targeting functions. The addition of 2,4-ε-dinitrophenol-L-lysine (DNP as a FcεRI targeting ligand and 4-mercaptopyridine (4-MPy as a pH-sensing ligand enables spatial and temporal monitoring of FcεRI receptors and their pH environment within the endocytic pathway. To ensure reliability, the sensor is calibrated in vivo using the ionophore nigericin and standard buffer solutions to equilibrate the external [H+] concentration with that of the cell compartments. This review highlights the nanosensors, ability to traffic and respond to pH of receptor-bound nanosensors (1 at physiological temperature (37°C versus room temperature (25°C, (2 after pharmacological treatment with bafilomycin, an H+ ATPase pump inhibitor, or amiloride, an inhibitor of Na+/H+ exchange, and (3 in response to both temperature and pharmacological treatment. Whole-cell, time lapse images are demonstrated to show the ability to transform live cells into dynamic laboratories to monitor temporal and spatial endosomal pH. The versatility of these probes shows promise for future applications relevant to intracellular trafficking and intelligent drug design.

  5. CellDyM: A room temperature operating cryogenic cell for the dynamic monitoring of snow metamorphism by time-lapse X-ray microtomography

    Science.gov (United States)

    Calonne, N.; Flin, F.; Lesaffre, B.; Dufour, A.; Roulle, J.; Puglièse, P.; Philip, A.; Lahoucine, F.; Geindreau, C.; Panel, J.-M.; Roscoat, S. Rolland; Charrier, P.

    2015-05-01

    Monitoring the time evolution of snow microstructure in 3-D is crucial for a better understanding of snow metamorphism. We, therefore, designed a cryogenic cell that precisely controls the experimental conditions of a sample while it is scanned by X-ray tomography. Based on a thermoelectrical regulation and a vacuum insulation, the cell operates at room temperature. It is, thus, adaptable to diverse scanners, offering advantages in terms of imaging techniques, resolution, and speed. Three-dimensional time-lapse series were obtained under equitemperature and temperature gradient conditions at a 7.8 μm precision. The typical features of each metamorphism and the anisotropic faceting behavior between the basal and prismatic planes, known to occur close to -2°C, were observed in less than 30 h. These results are consistent with the temperature fields expected from heat conduction simulations through the cell. They confirm the cell's accuracy and the interest of relatively short periods to study snow metamorphism.

  6. Time-lapse camera observations of gas piston activity at Pu‘u ‘Ō‘ō, Kīlauea volcano, Hawai‘i

    Science.gov (United States)

    Orr, Tim R.; Rea, James

    2012-01-01

    Gas pistoning is a type of eruptive behavior described first at Kīlauea volcano and characterized by the (commonly) cyclic rise and fall of the lava surface within a volcanic vent or lava lake. Though recognized for decades, its cause continues to be debated, and determining why and when it occurs has important implications for understanding vesiculation and outgassing processes at basaltic volcanoes. Here, we describe gas piston activity that occurred at the Pu‘u ‘Ō‘ō cone, in Kīlauea’s east rift zone, during June 2006. Direct, detailed measurements of lava level, made from time-lapse camera images captured at close range, show that the gas pistons during the study period lasted from 2 to 60 min, had volumes ranging from 14 to 104 m3, displayed a slowing rise rate of the lava surface, and had an average gas release duration of 49 s. Our data are inconsistent with gas pistoning models that invoke gas slug rise or a dynamic pressure balance but are compatible with models which appeal to gas accumulation and loss near the top of the lava column, possibly through the generation and collapse of a foam layer.

  7. Electromagnetically induced two-photon transparency in rubidium atoms

    International Nuclear Information System (INIS)

    Wang, D.; Gao, J.Y.; Xu, J.H.; Bassani, F.; La Rocca, G.C.; Salerno Univ.

    2001-01-01

    We present an experimental demonstration of electromagnetically induced two-photon transparency (EITT) in room temperature rubidium vapor. The 8S 1/2 to 5P 1/2 fluorescence is used to monitor the 5S 1/2 (F = 3) to 8S 1/2 (F = 3) two-photon absorption near resonance with the intermediate state 5P 3/2 . A controlling pump laser beam is employed to coherently couple the 5P 3/2 and 5D 5/2 states, thus producing two dressed intermediate states which give rise to destructive interference in the two-photon transition. An induced two-photon transparency of about 80% has been obtained at resonance; our experimental findings are in good agreement with the general theory of Agarwal et al. (1996), when the appropriate spectroscopic parameters are used. (orig.)

  8. Standard Model Higgs decay for two Photons in CMS

    CERN Multimedia

    Daniel Denegri

    2000-01-01

    Simulated two-photon mass distribution for SM Higgs and expected background in the CMS PbW04 crystal calorimeter for an integrated luminosity of 10 . 5 pb-1, with detailed simulation of calorimeter response.

  9. All-optical femtosecond switch using two-photon absorption

    International Nuclear Information System (INIS)

    Yavuz, D. D.

    2006-01-01

    Utilizing a two-photon absorption scheme in an alkali-metal vapor cell, we suggest a technique where a strong laser beam switches off another laser beam of different wavelength in femtosecond time scales

  10. A time-lapse gravity survey of the Coso geothermal field, China Lake Naval Air Weapons Station, California

    Science.gov (United States)

    Phelps, Geoffrey; Cronkite-Ratcliff, Collin; Blake, Kelly

    2018-04-19

    We have conducted a gravity survey of the Coso geothermal field to continue the time-lapse gravity study of the area initiated in 1991. In this report, we outline a method of processing the gravity data that minimizes the random errors and instrument bias introduced into the data by the Scintrex CG-5 relative gravimeters that were used. After processing, the standard deviation of the data was estimated to be ±13 microGals. These data reveal that the negative gravity anomaly over the Coso geothermal field, centered on gravity station CER1, is continuing to increase in magnitude over time. Preliminary modeling indicates that water-table drawdown at the location of CER1 is between 65 and 326 meters over the last two decades. We note, however, that several assumptions on which the model results depend, such as constant elevation and free-water level over the study period, still require verification.

  11. Visualization of conduit-matrix conductivity differences in a karst aquifer using time-lapse electrical resistivity

    Science.gov (United States)

    Meyerhoff, Steven B.; Karaoulis, Marios; Fiebig, Florian; Maxwell, Reed M.; Revil, André; Martin, Jonathan B.; Graham, Wendy D.

    2012-12-01

    In the karstic upper Floridan aquifer, surface water flows into conduits of the groundwater system and may exchange with water in the aquifer matrix. This exchange has been hypothesized to occur based on differences in discharge at the Santa Fe River Sink-Rise system, north central Florida, but has yet to be visualized using any geophysical techniques. Using electrical resistivity tomography, we conducted a time-lapse study at two locations with mapped conduits connecting the Santa Fe River Sink to the Santa Fe River Rise to study changes of electrical conductivity during times of varying discharge over a six-week period. Our results show conductivity differences between matrix, conduit changes in resistivity occurring through time at the locations of mapped karst conduits, and changes in electrical conductivity during rainfall infiltration. These observations provide insight into time scales and matrix conduit conductivity differences, illustrating how surface water flow recharged to conduits may flow in a groundwater system in a karst aquifer.

  12. Development and setting of a time-lapse video camera system for the Antarctic lake observation

    Directory of Open Access Journals (Sweden)

    Sakae Kudoh

    2010-11-01

    Full Text Available A submersible video camera system, which aimed to record the growth image of aquatic vegetation in Antarctic lakes for one year, was manufactured. The system consisted of a video camera, a programmable controller unit, a lens-cleaning wiper with a submersible motor, LED lights, and a lithium ion battery unit. Changes of video camera (High Vision System and modification of the lens-cleaning wiper allowed higher sensitivity and clearer recording images compared to the previous submersible video without increasing the power consumption. This system was set on the lake floor in Lake Naga Ike (a tentative name in Skarvsnes in Soya Coast, during the summer activity of the 51th Japanese Antarctic Research Expedition. Interval record of underwater visual image for one year have been started by our diving operation.

  13. Mass distribution for the two-photon channel

    CERN Multimedia

    ATLAS, collaboration

    2012-01-01

    Mass distribution for the two-photon channel. The strongest evidence for this new particle comes from analysis of events containing two photons. The smooth dotted line traces the measured background from known processes. The solid line traces a statistical fit to the signal plus background. The new particle appears as the excess around 126.5 GeV. The full analysis concludes that the probability of such a peak is three chances in a million.

  14. Two-photon quantum interference in a Michelson interferometer

    International Nuclear Information System (INIS)

    Odate, Satoru; Wang Haibo; Kobayashi, Takayoshi

    2005-01-01

    We have observed two-photon quantum interference in a Michelson interferometer. For the first time, we experimentally demonstrated two-photon quantum interference patterns, which show the transition from nonsubwavelength interference fringes to the general subwavelength interference. At the same time, a photon bunching effect was also shown by a postselection. The |1, 1> state with a single photon in a mode corresponding to each arm of the interferometer was exclusively postselected by using path difference between two arms

  15. Time-lapse ERT and DTS for seasonal and short-term monitoring of an alpine river hyporheic zone

    Science.gov (United States)

    Boaga, Jacopo; Laura, Busato; Mariateresa, Perri; Giorgio, Cassiani

    2016-04-01

    The hyporheic zone (HZ) is the area located beneath and adjacent to rivers and streams, where the interactions between surface water and groundwater take place. This complex physical domain allows the transport of several substances from a stream to the unconfined aquifer below, and vice versa, thus playing a fundamental role in the river ecosystem. The importance of the hyporheic zone makes its characterization a goal shared by several disciplines, which range from applied geophysics to biogeochemistry, from hydraulics to ecology. The frontier field of HZ characterization stays in applied non-invasive methodologies as Electrical Resistivity Tomography - ERT - and Distributed Temperature Sensing - DTS. ERT is commonly applied in cross-well configuration or with a superficial electrodes deployment while DTS is used in hydro-geophysics in the last decade, revealing a wide applicability to the typical issues of this field of study. DTS for hydro-geophysics studies is based on Raman scattering and employs heat as tracer and uses a fiber-optic cable to acquire temperature values. We applied both techniques for an alpine river case studies located in Val di Sole, TN, Italy. The collected measurements allow high-resolution characterization of the hyporheic zone, overcoming the critical problem of invasive measurements under riverbeds. In this work, we present the preliminary results regarding the characterization of the hyporheic zone of the alpine river obtained combining ERT and DTS time-lapse measurements. The data collection benefits from an innovative instrumentation deployment, which consists of both an ERT multicore cable and a DTS fiber-optic located in two separated boreholes drilled 5m under the watercourse and perpendicular to it. In particular we present the first year monitoring results and a short time-lapse monitoring experiment conducted during summer 2015. The site and the results here described are part of the EU FP7 CLIMB (Climate Induced Changes on the

  16. Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography.

    Science.gov (United States)

    Payne, D; Flaherty, S P; Barry, M F; Matthews, C D

    1997-03-01

    In this study, we have used time-lapse video cinematography to study fertilization in 50 human oocytes that had undergone intracytoplasmic sperm injection (ICSI). Time-lapse recording commenced shortly after ICSI and proceeded for 17-20 h. Oocytes were cultured in an environmental chamber which was maintained under standard culture conditions. Overall, 38 oocytes (76%) were fertilized normally, and the fertilization rate and embryo quality were not significantly different from 487 sibling oocytes cultured in a conventional incubator. Normal fertilization followed a defined course of events, although the timing of these events varied markedly between oocytes. In 35 of the 38 fertilized oocytes (92%), there were circular waves of granulation within the ooplasm which had a periodicity of 20-53 min. The sperm head decondensed during this granulation phase. The second polar body was then extruded, and this was followed by the central formation of the male pronucleus. The female pronucleus formed in the cytoplasm adjacent to the second polar body at the same time as, or slightly after, the male pronucleus, and was subsequently drawn towards the male pronucleus until the two abutted. Both pronuclei then increased in size, the nucleoli moved around within the pronuclei and some nucleoli coalesced. During pronuclear growth, the organelles contracted from the cortex towards the centre of the oocyte, leaving a clear cortical zone. The oocyte decreased in diameter from 112 to 106 microm (P cinematography is an excellent tool for studying fertilization and early embryo development, and have demonstrated that human fertilization comprises numerous complex dynamic events.

  17. Neural network control of focal position during time-lapse microscopy of cells.

    Science.gov (United States)

    Wei, Ling; Roberts, Elijah

    2018-05-09

    Live-cell microscopy is quickly becoming an indispensable technique for studying the dynamics of cellular processes. Maintaining the specimen in focus during image acquisition is crucial for high-throughput applications, especially for long experiments or when a large sample is being continuously scanned. Automated focus control methods are often expensive, imperfect, or ill-adapted to a specific application and are a bottleneck for widespread adoption of high-throughput, live-cell imaging. Here, we demonstrate a neural network approach for automatically maintaining focus during bright-field microscopy. Z-stacks of yeast cells growing in a microfluidic device were collected and used to train a convolutional neural network to classify images according to their z-position. We studied the effect on prediction accuracy of the various hyperparameters of the neural network, including downsampling, batch size, and z-bin resolution. The network was able to predict the z-position of an image with ±1 μm accuracy, outperforming human annotators. Finally, we used our neural network to control microscope focus in real-time during a 24 hour growth experiment. The method robustly maintained the correct focal position compensating for 40 μm of focal drift and was insensitive to changes in the field of view. About ~100 annotated z-stacks were required to train the network making our method quite practical for custom autofocus applications.

  18. Two-Photon-Absorption Scheme for Optical Beam Tracking

    Science.gov (United States)

    Ortiz, Gerardo G.; Farr, William H.

    2011-01-01

    A new optical beam tracking approach for free-space optical communication links using two-photon absorption (TPA) in a high-bandgap detector material was demonstrated. This tracking scheme is part of the canonical architecture described in the preceding article. TPA is used to track a long-wavelength transmit laser while direct absorption on the same sensor simultaneously tracks a shorter-wavelength beacon. The TPA responsivity was measured for silicon using a PIN photodiode at a laser beacon wavelength of 1,550 nm. As expected, the responsivity shows a linear dependence with incident power level. The responsivity slope is 4.5 x 10(exp -7) A/W2. Also, optical beam spots from the 1,550-nm laser beacon were characterized on commercial charge coupled device (CCD) and complementary metal-oxide semiconductor (CMOS) imagers with as little as 13.7 microWatts of optical power (see figure). This new tracker technology offers an innovative solution to reduce system complexity, improve transmit/receive isolation, improve optical efficiency, improve signal-to-noise ratio (SNR), and reduce cost for free-space optical communications transceivers.

  19. Examining the information content of time-lapse crosshole GPR data collected under different infiltration conditions to estimate unsaturated soil hydraulic properties

    DEFF Research Database (Denmark)

    Scholer, M.; Irving, J.; Zibar, Majken Caroline Looms

    2013-01-01

    Time-lapse geophysical data acquired during transient hydrological experiments are being increasingly employed to estimate subsurface hydraulic properties at the field scale. In particular, crosshole ground-penetrating radar (GPR) data, collected while water infiltrates into the subsurface either...... by natural or artificial means, have been demonstrated in a number of studies to contain valuable information concerning the hydraulic properties of the unsaturated zone. Previous work in this domain has considered a variety of infiltration conditions and different amounts of time-lapse GPR data...... of time-lapse zero-offset-profile (ZOP) GPR traveltime data, collected under three different infiltration conditions, for the estimation of van Genuchten–Mualem (VGM) parameters in a layered subsurface medium. Specifically, we systematically analyze synthetic and field GPR data acquired under natural...

  20. Embryonic lineage analysis using three-dimensional, time-lapse in-vivo fluorescent microscopy

    Science.gov (United States)

    Minden, Jonathan; Kam, Zvi; Agard, David A.; Sedat, John W.; Alberts, Bruce

    1990-08-01

    Drosophila melanogaster has become one of the most extensively studied organisms because of its amenability to genetic analysis. Unfortunately, the biochemistry and cell biology ofDrosophila has lagged behind. To this end we have been microinjecting fluorescently labelled proteins into the living embryo and observing the behavior of these proteins to determine their role in the cell cycle and development. Imaging of these fluorescent probes is an extremely important element to this form of analysis. We have taken advantage of the sensitivity and well behaved characteristics of the charge coupled device (CCD) camera in conjunction with digital image enhancement schemes to produce highly accurate images of these fluorescent probes in vivo. One of our major goals is to produce a detailed map of cell fate so that we can understand how fate is determined and maintained. In order produce such a detailed map, protocols for following the movements and mitotic behavior of a large number of cells in three dimensions over relatively long periods of time were developed. We will present our results using fluorescently labelled histone proteins as a marker for nuclear location1. In addition, we will also present our initial results using a photoactivatable analog of fluorescein to mark single cells so that their long range fate can be unambiguously determined.

  1. Three-dimensional time-lapse velocity tomography of an underground longwall panel

    Energy Technology Data Exchange (ETDEWEB)

    Luxbacher, K.; Westman, E.; Swanson, P.; Karfakis, M. [Virginia Tech., Blacksburg, VA (United States). Dept. of Mining & Minerals Engineering

    2008-06-15

    Three-dimensional velocity tomograms were generated to image the stress redistribution around an underground coal longwall panel to produce a better understanding of the mechanisms that lead to ground failure, especially rockbursts. Mining-induced microseismic events provided passive sources for the three-dimensional velocity tomography. Surface-mounted geophones monitored microseismic activity for 18 days. Eighteen tomograms were generated and high-velocity regions correlated with high abutment stresses predicted by numerical modeling. Additionally, the high-velocity regions were observed to redistribute as the longwall face retreated, indicating that velocity tomography may be an appropriate technology for monitoring stress redistribution in underground mines.

  2. Time-lapse electrical resistivity anomalies due to contaminant transport around landfills

    Directory of Open Access Journals (Sweden)

    J. Yang

    2007-06-01

    Full Text Available The extent of landfill leachate can be delineated by geo-electrical imaging as a response to the varying electrical resistivity in the contaminated area. This research was based on a combination of hydrogeological numerical simulation followed by geophysical forward and inversion modeling performed to evaluate the migration of a contaminant plume from a landfill. As a first step, groundwater flow and contaminant transport was simulated using the finite elements numerical modeling software FEFLOW. The extent of the contaminant plume was acquired through a hydrogeological model depicting the distributions of leachate concentration in the system. Next, based on the empirical relationship between the concentration and electrical conductivity of the leachate in the porous media, the corresponding geo-electrical structure was derived from the hydrogeological model. Finally, forward and inversion computations of geo-electrical anomalies were performed using the finite difference numerical modeling software DCIP2D/DCIP3D. The image obtained by geophysical inversion of the electric data was expected to be consistent with the initial hydrogeological model, as described by the distribution of leachate concentration. Numerical case studies were conducted for various geological conditions, hydraulic parameters and electrode arrays, from which conclusions were drawn regarding the suitability of the methodology to assess simple to more complex geo-electrical models. Thus, optimal mapping and monitoring configurations were determined.

  3. Sensing for intracellular thiols by water-insoluble two-photon fluorescent probe incorporating nanogel

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Xudong; Zhang, Xin; Wang, Shuangqing; Li, Shayu [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Hu, Rui, E-mail: hurui@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Li, Yi, E-mail: yili@mail.ipc.ac.cn [Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190 (China); Yang, Guoqiang, E-mail: gqyang@iccas.ac.cn [Beijing National Laboratory for Molecular Sciences, Key laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190 (China)

    2015-04-15

    Highlights: • A novel “turn-on” two-photon fluorescent probe based on a π-conjugated triarylboron luminogen was designed and synthesized. • Fast, selective and sensitive detection of biothiols in 100% aqueous solution by simply loaded on a nanogel. • Single-photon and two-photon fluorescent bioimaging of biothiols in NIH/3T3 fibroblasts. - Abstract: A novel “turn-on” two-photon fluorescent probe containing a π-conjugated triarylboron luminogen and a maleimide moiety DMDP-M based on the photo-induced electron transfer (PET) mechanism for biothiol detection was designed and synthesized. By simply loading the hydrophobic DMDP-M on a cross-linked Pluronic{sup ®} F127 nanogel (CL-F127), a probing system DMDP-M/CL-F127 was established, which shows quick response, high selectivity and sensitivity to cysteine (Cys), homocysteine (Hcy) and glutathione (GSH) in aqueous phase. The DMDP-M/CL-F127 system presented the fastest response to Cys with a rate constant of 0.56 min{sup −1}, and the detection limit to Cys was calculated to be as low as 0.18 μM. The DMDP-M/CL-F127 system has been successfully applied to the fluorescence imaging of biothiols in NIH/3T3 fibroblasts either with single-photon or two-photon excitation because of its high biocompatibility and cell-membrane permeability. The present work provides a general, simple and efficient strategy for the application of hydrophobic molecules to sensing biothiols in aqueous phase, and a novel sensing system for intracellular biothiols fitted for both single-photon and two-photon fluorescence imaging.

  4. An integrated single- and two-photon non-diffracting light-sheet microscope

    Science.gov (United States)

    Lau, Sze Cheung; Chiu, Hoi Chun; Zhao, Luwei; Zhao, Teng; Loy, M. M. T.; Du, Shengwang

    2018-04-01

    We describe a fluorescence optical microscope with both single-photon and two-photon non-diffracting light-sheet excitations for large volume imaging. With a special design to accommodate two different wavelength ranges (visible: 400-700 nm and near infrared: 800-1200 nm), we combine the line-Bessel sheet (LBS, for single-photon excitation) and the scanning Bessel beam (SBB, for two-photon excitation) light sheet together in a single microscope setup. For a transparent thin sample where the scattering can be ignored, the LBS single-photon excitation is the optimal imaging solution. When the light scattering becomes significant for a deep-cell or deep-tissue imaging, we use SBB light-sheet two-photon excitation with a longer wavelength. We achieved nearly identical lateral/axial resolution of about 350/270 nm for both imagings. This integrated light-sheet microscope may have a wide application for live-cell and live-tissue three-dimensional high-speed imaging.

  5. Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

    Science.gov (United States)

    Kwok, Sheldon J. J.; Kuznetsov, Ivan A.; Kim, Moonseok; Choi, Myunghwan; Scarcelli, Giuliano; Yun, Seok-Hyun

    2017-02-01

    Two-photon polymerization and crosslinking are commonly used methods for microfabrication of three-dimensional structures with applications spanning from photonic microdevices, drug delivery systems, to cellular scaffolds. However, the use of two-photon processes for precise, internal modification of biological tissues has not yet been reported. One of the major challenges has been a lack of appropriate tools to monitor and characterize crosslinked regions nondestructively. Here, we demonstrate spatially selective two-photon collagen crosslinking (2P-CXL) in intact tissue for the first time. Using riboflavin photosensitizer and femtosecond laser irradiation, we crosslinked a small volume of tissue within animal corneas. Collagen fiber orientations and photobleaching were characterized by second harmonic generation and two-photon fluorescence imaging, respectively. Using confocal Brillouin microscopy, we measured local changes in longitudinal mechanical moduli and visualized the cross-linked pattern without perturbing surrounding non-irradiated regions. 2P-CXL-induced tissue stiffening was comparable to that achieved with conventional one-photon CXL. Our results demonstrate the ability to selectively stiffen biological tissue in situ at high spatial resolution, with broad implications in ophthalmology, laser surgery, and tissue engineering.

  6. Polarized two-photon photoselection in EGFP: Theory and experiment.

    Science.gov (United States)

    Masters, T A; Marsh, R J; Blacker, T S; Armoogum, D A; Larijani, B; Bain, A J

    2018-04-07

    In this work, we present a complete theoretical description of the excited state order created by two-photon photoselection from an isotropic ground state; this encompasses both the conventionally measured quadrupolar (K = 2) and the "hidden" degree of hexadecapolar (K = 4) transition dipole alignment, their dependence on the two-photon transition tensor and emission transition dipole moment orientation. Linearly and circularly polarized two-photon absorption (TPA) and time-resolved single- and two-photon fluorescence anisotropy measurements are used to determine the structure of the transition tensor in the deprotonated form of enhanced green fluorescent protein. For excitation wavelengths between 800 nm and 900 nm, TPA is best described by a single element, almost completely diagonal, two-dimensional (planar) transition tensor whose principal axis is collinear to that of the single-photon S 0 → S 1 transition moment. These observations are in accordance with assignments of the near-infrared two-photon absorption band in fluorescent proteins to a vibronically enhanced S 0 → S 1 transition.

  7. Polarized two-photon photoselection in EGFP: Theory and experiment

    Science.gov (United States)

    Masters, T. A.; Marsh, R. J.; Blacker, T. S.; Armoogum, D. A.; Larijani, B.; Bain, A. J.

    2018-04-01

    In this work, we present a complete theoretical description of the excited state order created by two-photon photoselection from an isotropic ground state; this encompasses both the conventionally measured quadrupolar (K = 2) and the "hidden" degree of hexadecapolar (K = 4) transition dipole alignment, their dependence on the two-photon transition tensor and emission transition dipole moment orientation. Linearly and circularly polarized two-photon absorption (TPA) and time-resolved single- and two-photon fluorescence anisotropy measurements are used to determine the structure of the transition tensor in the deprotonated form of enhanced green fluorescent protein. For excitation wavelengths between 800 nm and 900 nm, TPA is best described by a single element, almost completely diagonal, two-dimensional (planar) transition tensor whose principal axis is collinear to that of the single-photon S0 → S1 transition moment. These observations are in accordance with assignments of the near-infrared two-photon absorption band in fluorescent proteins to a vibronically enhanced S0 → S1 transition.

  8. Time-lapse gravity and levelling in the sinkhole-endangered urban area of Bad Frankenhausen, Germany

    Science.gov (United States)

    Kobe, Martin; Gabriel, Gerald; Weise, Adelheid; Krawczyk, Charlotte; Vogel, Detlef

    2017-04-01

    Sinkholes, resulting from subrosion in the subsurface, can reach diameters of several hundred meters and thus pose a severe hazard for infrastructure and inhabitants in urban areas. Subrosion is the leaching of readily-soluble rocks, such as rock salt, gypsum, anhydrite and limestone by ground or meteoric water and leads to mass transport and relocation. Two scenarios of sinkhole evolution are conceivable: First, the surface subsides continuously in order to compensate for the mass loss. Second, the mass relocation leads to development of subsurface cavities. If they reach a critical size and the cover layers are not supported anymore, the surface collapses abruptly. To improve the understanding of subrosion processes and the related surface deformation a case study is conducted in Bad Frankenhausen, Germany, where subrosion leaches the Zechstein evaporates of the Permian. One part of the study is to analyse the spatiotemporal development of sinkholes by applying time-lapse observations. Therefore, we established a monitoring network consisting of 15 gravity and additional levelling points covering the main sinkhole areas in the city centre. In March 2014, the baseline survey was carried out. Since then, quarterly measurement campaigns are performed. In each campaign four different gravity meters are used to collect a statistical significant amount of data and to control the plausibility of our data. The gravity measurements are complemented by levelling surveys. The rectification of the time-lapse gravity data comprises the correction for jumps and systematic errors, as well as for well calculable influences, such as earth tides and air pressure changes. Furthermore, special interest was applied to seasonal changes of hydrological parameters such as soil moisture or groundwater level. We found the hydrological influence to be in the single digit up to the lower two-digit µGal range, depending on the season and the station. The standard deviations of the adjusted

  9. Interference of two photons in parametric down conversion

    International Nuclear Information System (INIS)

    Ghosh, R.; Hong, C.K.; Ou, Z.Y.; Mandel, L.

    1986-01-01

    A theoretical treatment is given of the process in which the two photons produced simultaneously in the parametric frequency splitting of light are allowed to interfere. It is shown that, while there is no interference in the usual sense involving quantities that are of the second order in the field, fourth-order interference effects are present. These may be revealed by measuring the joint probability of detecting two photons at two points x,x' in the interference plane with photoelectric detectors as a function of the separation x-x'. The probability exhibits a cosine modulation with x-x', with visibility that can approach 100%, even though the integration time in the experiment may greatly exceed the reciprocal bandwidth of the photons. The interference effect has a nonclassical origin and implies a violation of local realism in the highly correlated two-photon state

  10. Measurement of wavefront aberrations in cortex and peripheral nerve using a two-photon excitation guidestar

    Science.gov (United States)

    Futia, Gregory L.; Fontaine, Arjun; McCullough, Connor; Ozbay, Baris N.; George, Nickolas M.; Caldwell, John; Restrepo, Diego; Weir, Richard; Gibson, Emily A.

    2018-02-01

    Neural-machine interfaces using optogenetics are of interest due to their minimal invasiveness and potential for parallel read in and read out of activity. One possible biological target for such an interface is the peripheral nerve, where axonlevel imaging or stimulation could greatly improve interfacing with artificial limbs or enable neuron/fascicle level neuromodulation in the vagus nerve. Two-photon imaging has been successful in imaging brain activity using genetically encoded calcium or voltage indicators, but in the peripheral nerve, this is severely limited by scattering and aberrations from myelin. We employ a Shack-Hartman wavefront sensor and two-photon excitation guidestar to quantify optical scattering and aberrations in peripheral nerves and cortex. The sciatic and vagus nerves, and cortex from a ChAT-Cre ChR-eYFP transgenic mouse were excised and imaged directly. In peripheral nerves, defocus was the strongest aberration followed by astigmatism and coma. Peripheral nerve had orders of magnitude higher aberration compared with cortex. These results point to the potential of adaptive optics for increasing the depth of two-photon access into peripheral nerves.

  11. Doppler-free two-photon excitation of 238U

    International Nuclear Information System (INIS)

    Hodgkinson, D.P.; Wort, D.J.H.

    1981-04-01

    A theory of resonantly enhanced two-photon absorption is presented and tested in a number of experiments in which 238 U vapour is excited by two continuous wave dye lasers. Good quantitative agreement between theory and experiment is found. In particular the central prediction of the theory, that antiparallel laser beams of modest intensity can pump an appreciable fraction of the Maxwell velocity distribution, has been checked directly by measuring the spectral width of the fluorescence from the two-photon excited level. (author)

  12. Time-lapse culture with morphokinetic embryo selection improves pregnancy and live birth chances and reduces early pregnancy loss: a meta-analysis.

    Science.gov (United States)

    Pribenszky, Csaba; Nilselid, Anna-Maria; Montag, Markus

    2017-11-01

    Embryo evaluation and selection is fundamental in clinical IVF. Time-lapse follow-up of embryo development comprises undisturbed culture and the application of the visual information to support embryo evaluation. A meta-analysis of randomized controlled trials was carried out to study whether time-lapse monitoring with the prospective use of a morphokinetic algorithm for selection of embryos improves overall clinical outcome (pregnancy, early pregnancy loss, stillbirth and live birth rate) compared with embryo selection based on single time-point morphology in IVF cycles. The meta-analysis of five randomized controlled trials (n = 1637) showed that the application of time-lapse monitoring was associated with a significantly higher ongoing clinical pregnancy rate (51.0% versus 39.9%), with a pooled odds ratio of 1.542 (P loss (15.3% versus 21.3%; OR: 0.662; P = 0.019) and a significantly increased live birth rate (44.2% versus 31.3%; OR 1.668; P = 0.009). Difference in stillbirth was not significant between groups (4.7% versus 2.4%). Quality of the evidence was moderate to low owing to inconsistencies across the studies. Selective application and variability were also limitations. Although time-lapse is shown to significantly improve overall clinical outcome, further high-quality evidence is needed before universal conclusions can be drawn. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  13. Monitoring channel head erosion processes in response to an artificially induced abrupt base level change using time-lapse photography 2301

    Science.gov (United States)

    Headcut and channel extension in response to an abrupt base level change in 2004 of approximately 1m was studied in a 1.29 ha semiarid headwater drainage on the Walnut Gulch Experimental Watershed (WGEW) in southeastern Arizona, USA. Field observations and time-lapse photography were coupled with hy...

  14. Assessing Uncertainty and Repeatability in Time-Lapse VSP Monitoring of CO2 Injection in a Brine Aquifer, Frio Formation, Texas (A Case Study)

    Energy Technology Data Exchange (ETDEWEB)

    Nazari, Siamak [Univ. of California, Berkeley, CA (United States). Dept. of Civil and Environmental Engineering; Daley, Thomas M. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth Sciences Division

    2013-02-07

    This study was done to assess the repeatability and uncertainty of time-lapse VSP response to CO2 injection in the Frio formation near Houston Texas. A work flow was built to assess the effect of time-lapse injected CO2 into two Frio brine reservoir intervals, the ‘C’ sand (Frio1) and the ‘Blue sand’ (Frio2). The time-lapse seismic amplitude variations with sensor depth for both reservoirs Frio1 and Frio2 were computed by subtracting the seismic response of the base survey from each of the two monitor seismic surveys. Source site 1 has been considered as one of the best sites for evaluating the time-lapse response after injection. For site 1, the computed timelapse NRMS levels after processing had been compared to the estimated time-lapse NRMS level before processing for different control reflectors, and for brine aquifers Frio1, and Frio2 to quantify detectability of amplitude difference. As the main interest is to analyze the time-lapse amplitude variations, different scenarios have been considered. Three different survey scenarios were considered: the base survey which was performed before injection, monitor1 performed after the first injection operation, and monitor2 which was after the second injection. The first scenario was base-monitor1, the second was basemonitor2, and the third was monitor1-monitor2. We considered three ‘control’ reflections above the Frio to assist removal of overburden changes, and concluded that third control reflector (CR3) is the most favorable for the first scenario in terms of NRMS response, and first control reflector (CR1) is the most favorable for the second and third scenarios in terms of NRMS response. The NRMS parameter is shown to be a useful measure to assess the effect of processing on time-lapse data. The overall NRMS for the Frio VSP data set was found to be in the range of 30% to 80% following basic processing. This could be considered as an estimated baseline in assessing the utility

  15. Fighting for territories: time-lapse analysis of dental pulp and dental follicle stem cells in co-culture reveals specific migratory capabilities

    Directory of Open Access Journals (Sweden)

    C Schiraldi

    2012-11-01

    Full Text Available Stem cell migration is a critical step during the repair of damaged tissues. In order to achieve appropriate cell-based therapies for tooth and periodontal ligament repair it is necessary first to understand the dynamics of tissue-specific stem cell populations such as dental pulp stem cells (DPSC and dental follicle stem cells (DFSC. Using time-lapse imaging, we analysed migratory and proliferative capabilities of these two human stem cell lines in vitro. When cultured alone, both DPSC and DFSC exhibited low and irregular migration profiles. In co-cultures, DFSC, but not DPSC, spectacularly increased their migration activity and velocity. DFSC rapidly surrounded the DPSC, thus resembling the in vivo developmental process, where follicle cells encircle both dental epithelium and pulp. Cell morphology was dependent on the culture conditions (mono-culture or co-culture and changed over time. Regulatory genes involved in dental cell migration and differentiation such as TWIST1, MSX1, RUNX2, SFRP1 and ADAM28, were also evaluated in co-cultures. MSX1 up-regulation indicates that DPSC and DFSC retain their odontogenic potential. However, DPSC lose their capacity to differentiate into odontoblasts in the presence of DFSC, as suggested by RUNX2 up-regulation and TWIST1 down-regulation. In contrast, the unchanged levels of SFRP1 expression suggest that DFSC retain their potential to form periodontal tissues even in the presence of DPSC. These findings demonstrate that stem cells behave differently according to their environment, retain their genetic memory, and compete with each other to acquire the appropriate territory. Understanding the mechanisms involved in stem cell migration may lead to new therapeutic approaches for tooth repair.

  16. The turbulent life of juvenile icebergs: Observations from an array of high-rate time-lapse cameras in LeConte Bay, Alaska

    Science.gov (United States)

    Kienholz, C.; Amundson, J. M.; Jackson, R. H.; Motyka, R. J.; Nash, J. D.; Sutherland, D.

    2017-12-01

    Tidewater glacier behavior is driven by poorly understood processes occurring at the ice-ocean interface, including sedimentation and erosion, iceberg calving, and submarine melting. These processes are inherently difficult to observe, calling for innovative field techniques and numerical models. As part of a multi-year field effort to constrain ocean-glacier heat and mass exchange, we deployed an array of high-rate time-lapse cameras (sampling intervals between 15 seconds and 2 minutes) to monitor the terminus of LeConte Glacier and its proglacial fjord. The camera array has operated continuously for more than a year. Our high sampling rates enable tracking of iceberg motion with optical flow algorithms, which have been used widely in computer vision but less so in glaciology and oceanography. Such algorithms track individual features (e.g., corners of icebergs), which is ideal for iceberg-rich fjords, where motion can vary substantially over short temporal and spatial scales (e.g., due to complex surface currents or different iceberg sizes). We process our data to quantify subdaily to seasonal patterns in surface currents and relate them to forcing from tides, wind, and glacier runoff. Flow is most variable close to the glacier terminus due to frequent calving events and turbulent plume dynamics. Farther down fjord, more consistent patterns emerge, driven by tides, wind, and runoff and altered by fjord geometry. Our tracking results compare favorably to and complement our Acoustic Doppler Current Profiler measurements from boats and moorings. Given their high spatial and temporal resolution, our observations will place important surface constraints on forthcoming hydrodynamic modeling efforts. The deployment of the cameras in a harsh environment and the corresponding image processing provided an opportunity to test hardware and software thoroughly, which will prove useful for similar systems at other glaciers.

  17. Spatial and temporal variations of thaw layer thickness and its controlling factors identified using time-lapse electrical resistivity tomography and hydro-thermal modeling

    Science.gov (United States)

    Tran, Anh Phuong; Dafflon, Baptiste; Bisht, Gautam; Hubbard, Susan S.

    2018-06-01

    Quantitative understanding of controls on thaw layer thickness (TLT) dynamics in the Arctic peninsula is essential for predictive understanding of permafrost degradation feedbacks to global warming and hydrobiochemical processes. This study jointly interprets electrical resistivity tomography (ERT) measurements and hydro-thermal numerical simulation results to assess spatiotemporal variations of TLT and to determine its controlling factors in Barrow, Alaska. Time-lapse ERT measurements along a 35-m transect were autonomously collected from 2013 to 2015 and inverted to obtain soil electrical resistivity. Based on several probe-based TLT measurements and co-located soil electrical resistivity, we estimated the electrical resistivity thresholds associated with the boundary between the thaw layer and permafrost using a grid search optimization algorithm. Then, we used the obtained thresholds to derive the TLT from all soil electrical resistivity images. The spatiotemporal analysis of the ERT-derived TLT shows that the TLT at high-centered polygons (HCPs) is smaller than that at low-centered polygons (LCPs), and that both thawing and freezing occur earlier at the HCPs compared to the LCPs. In order to provide a physical explanation for dynamics in the thaw layer, we performed 1-D hydro-thermal simulations using the community land model (CLM). Simulation results showed that air temperature and precipitation jointly govern the temporal variations of TLT, while the topsoil organic content (SOC) and polygon morphology are responsible for its spatial variations. When the topsoil SOC and its thickness increase, TLT decreases. Meanwhile, at LCPs, a thicker snow layer and saturated soil contribute to a thicker TLT and extend the time needed for TLT to freeze and thaw. This research highlights the importance of combination of measurements and numerical modeling to improve our understanding spatiotemporal variations and key controls of TLT in cold regions.

  18. Vapor flux and recrystallization during dry snow metamorphism under a steady temperature gradient as observed by time-lapse micro-tomography

    Directory of Open Access Journals (Sweden)

    B. R. Pinzer

    2012-10-01

    Full Text Available Dry snow metamorphism under an external temperature gradient is the most common type of recrystallization of snow on the ground. The changes in snow microstructure modify the physical properties of snow, and therefore an understanding of this process is essential for many disciplines, from modeling the effects of snow on climate to assessing avalanche risk. We directly imaged the microstructural changes in snow during temperature gradient metamorphism (TGM under a constant gradient of 50 K m−1, using in situ time-lapse X-ray micro-tomography. This novel and non-destructive technique directly reveals the amount of ice that sublimates and is deposited during metamorphism, in addition to the exact locations of these phase changes. We calculated the average time that an ice volume stayed in place before it sublimated and found a characteristic residence time of 2–3 days. This means that most of the ice changes its phase from solid to vapor and back many times in a seasonal snowpack where similar temperature conditions can be found. Consistent with such a short timescale, we observed a mass turnover of up to 60% of the total ice mass per day. The concept of hand-to-hand transport for the water vapor flux describes the observed changes very well. However, we did not find evidence for a macroscopic vapor diffusion enhancement. The picture of {temperature gradient metamorphism} produced by directly observing the changing microstructure sheds light on the micro-physical processes and could help to improve models that predict the physical properties of snow.

  19. A derivative-free approach for the estimation of porosity and permeability using time-lapse seismic and production data

    International Nuclear Information System (INIS)

    Dadashpour, Mohsen; Kleppe, Jon; Landrø, Martin; Echeverria Ciaurri, David; Mukerji, Tapan

    2010-01-01

    In this study, we apply a derivative-free optimization algorithm to estimate porosity and permeability from time-lapse seismic data and production data from a real reservoir (Norne field). In some circumstances, obtaining gradient information (exact and/or approximate) can be problematic e.g. derivatives are not available from a commercial simulator, or results are needed within a very short time frame. Derivative-free optimization approaches can be very time consuming because they often require many simulations. Typically, one iteration roughly needs as many simulations as the number of optimization variables. In this work, we propose two ways to significantly increase the efficiency of an optimization methodology in model inversion problems. First, by principal component analysis we decrease the number of optimization variables while keeping geostatistical consistency, and second, noticing that some optimization methods are very amenable to being parallelized, we apply them within a distributed computing framework. If we combine all this, the model inversion approach can be robust, fairly efficient and very simple to implement. In this paper, we apply the methodology to two cases: a semi-synthetic model with noisy data, and a case based entirely on field data. The results show that the derivative-free approach presented is robust against noise in the data

  20. Comparative effects of ionizing radiation on cycle time and mitotic duration. A time-lapse cinematography study

    International Nuclear Information System (INIS)

    D'Hooghe, M.C.; Hemon, D.; Valleron, A.J.; Malaise, E.P.

    1980-01-01

    The effects of 60 Co γ rays on the length of the intermitotic period, the duration of mitosis, and the division probability of EMT6 cells have been studied in vitro using time-lapse cinematography. Irradiation increases the duration of the mitosis and of the cycle in comparable proportions: both parameters are practically doubled by a dose of 10 Gy. When daughters of irradiated cells die, the mitotic delay and lengthening of mitosis of their mother cells are longer than average. Mitotic delay and lengthening of mitosis depend on the age of cells at the moment of irradiation. The mitotic delay increases progressively when cells are irradiated during the first 8 h of their cycle (i.e., before the transition point), whereas mitosis is slightly prolonged. On the other hand, when the cells are irradiated after this transition point the mitotic delay decreases markedly, whereas the lengthening of mitosis increases sharply. These results tend to indicate that two different mechanisms are responsible for mitotic delay and prolongation of mitosis observed after irradiation

  1. Comparative effects of ionizing radiation on cycle time and mitotic duration. A time-lapse cinematography study

    Energy Technology Data Exchange (ETDEWEB)

    D' Hooghe, M.C. (Institut de Recherches sur le Cancer, Lille, France); Hemon, D.; Valleron, A.J.; Malaise, E.P.

    1980-03-01

    The effects of /sup 60/Co ..gamma.. rays on the length of the intermitotic period, the duration of mitosis, and the division probability of EMT6 cells have been studied in vitro using time-lapse cinematography. Irradiation increases the duration of the mitosis and of the cycle in comparable proportions: both parameters are practically doubled by a dose of 10 Gy. When daughters of irradiated cells die, the mitotic delay and lengthening of mitosis of their mother cells are longer than average. Mitotic delay and lengthening of mitosis depend on the age of cells at the moment of irradiation. The mitotic delay increases progressively when cells are irradiated during the first 8 h of their cycle (i.e., before the transition point), whereas mitosis is slightly prolonged. On the other hand, when the cells are irradiated after this transition point the mitotic delay decreases markedly, whereas the lengthening of mitosis increases sharply. These results tend to indicate that two different mechanisms are responsible for mitotic delay and prolongation of mitosis observed after irradiation.

  2. The sexual phase of the diatom Pseudo-nitzschia multistriata: cytological and time-lapse cinematography characterization.

    Science.gov (United States)

    Scalco, Eleonora; Amato, Alberto; Ferrante, Maria Immacolata; Montresor, Marina

    2016-11-01

    Pseudo-nitzschia is a thoroughly studied pennate diatom genus for ecological and biological reasons. Many species in this genus, including Pseudo-nitzschia multistriata, can produce domoic acid, a toxin responsible for amnesic shellfish poisoning. Physiological, phylogenetic and biological features of P. multistriata were studied extensively in the past. Life cycle stages, including the sexual phase, fundamental in diatoms to restore the maximum cell size and avoid miniaturization to death, have been well described for this species. P. multistriata is heterothallic; sexual reproduction is induced when strains of opposite mating type are mixed, and proceeds with cells producing two functionally anisogamous gametes each; however, detailed cytological information for this process is missing. By means of confocal laser scanning microscopy and nuclear staining, we followed the nuclear fate during meiosis, and using time-lapse cinematography, we timed every step of the sexual reproduction process from mate pairing to initial cell hatching. The present paper depicts cytological aspects during gametogenesis in P. multistriata, shedding light on the chloroplast behaviour during sexual reproduction, finely describing the timing of the sexual phases and providing reference data for further studies on the molecular control of this fundamental process.

  3. Precision two-photon spectroscopy of alkali elements

    Indian Academy of Sciences (India)

    effect is eliminated if the wave vector of the photons is ka = −kb, i.e., the two beams .... atomic cesium, and the metre is (indirectly) defined from the wavelength of .... plasma absorb radiation, the electrical circuit parameters may vary and this .... two-photon excitation followed by an ionization step in Ca [71], Sr [72] and Gd [73] ...

  4. Two-photon couplings of quarkonia with arbitrary JPC

    International Nuclear Information System (INIS)

    Barnes, T.; Tennessee Univ., Knoxville, TN

    1992-01-01

    We present theoretical results for the two-photon widths of relativistic quarkonium states with arbitrary angular momenta. These relativistic formulas are required to obtain reasonable agreement with the absolute scale of quarkonium decay rates to two photons, and have previously only been derived for spin-singlet q bar q states. We also evaluate these formulas numerically for ell ≤3 q = u, d states in a Coulomb-plus-linear q bar q potential model. Light-quark higher-ell and radially-excited q bar q states should be observable experimentally, as their two-photon widths are typically found to be ∼1 KeV. The radially-excited 1 S 0 higher-mass quarkonium states such as c bar c and b bar b should also be observable in γγ, but orbitally-excited c bar c states with ell>1 and b bar b states with ell>0 are expected to have very small two-photon widths. The helicity structure of the higher-ell q bar q couplings is predicted to be nontrivial, with both λ=0 and λ=2γγ final states contributing significantly; these results may be useful as signatures for q bar q states

  5. Two-photon optics of Bessel-Gaussian modes

    CSIR Research Space (South Africa)

    McLaren, M

    2013-09-01

    Full Text Available In this paper we consider geometrical two-photon optics of Bessel-Gaussian modes generated in spontaneous parametric down-conversion of a Gaussian pump beam. We provide a general theoretical expression for the orbital angular momentum (OAM) spectrum...

  6. Space-time description of the two-photon decay

    International Nuclear Information System (INIS)

    Hrasko, P.

    1981-09-01

    The time correlation of photons in a two-photon decay is shown to depend on the instantaneous nature of the wave-function collapse in an essential way so the latter hypothesis can be verified by the experimental study of these correlations. (author)

  7. Positron two-photon annihilation coincidence technique: difference mode

    Energy Technology Data Exchange (ETDEWEB)

    Karol, P J; Klobuchar, R L [Carnegie-Mellon Univ., Pittsburgh, Pa. (USA). Dept. of Chemistry

    1978-05-01

    A difference (or comparative) mode in the measurement of two-photon coincidences from positron or positronium annihilation has been developed. The method can be used to advantage, particularly in gases, in determining annihilation parameters such as quenching cross sections for low concentration strongly quenching chemical species in a reference medium which is relatively non-quenching.

  8. Two-photon absorption and two-photon circular dichroism of hexahelicene derivatives: a study of the effect of the nature of intramolecular charge transfer

    Czech Academy of Sciences Publication Activity Database

    Díaz, C.; Vesga, Y.; Echevarria, L.; Stará, Irena G.; Starý, Ivo; Anger, E.; Shen, C.; Moussa, M. E. S.; Vanthuyne, N.; Crassous, J.; Rizzo, A.; Hernández, F. E.

    2015-01-01

    Roč. 5, č. 23 (2015), s. 17429-17437 ISSN 2046-2069 Institutional support: RVO:61388963 Keywords : [6]helicene * two photon absorption * two photon CD Subject RIV: CC - Organic Chemistry Impact factor: 3.289, year: 2015

  9. 2D Time-lapse Resistivity Monitoring of an Organic Produced Gas Plume in a Landfill using ERT.

    Science.gov (United States)

    Amaral, N. D.; Mendonça, C. A.; Doherty, R.

    2014-12-01

    This project has the objective to study a landfill located on the margins of Tietê River, in São Paulo, Brazil, using the electroresistivity tomography method (ERT). Due to huge organic matter concentrations in the São Paulo Basin quaternary sediments, there is subsurface depth related biogas accumulation (CH4 and CO2), induced by anaerobic degradation of the organic matter. 2D resistivity sections were obtained from a test area since March 2012, a total of 7 databases, being the last one dated from October 2013. The studied line has the length of 56m, the electrode interval is of 2m. In addition, there are two boreholes along the line (one with 3 electrodes and the other one with 2) in order to improve data quality and precision. The boreholes also have a multi-level sampling system that indicates the fluid (gas or water) presence in relation to depth. With our results it was possible to map the gas plume position and its area of extension in the sections as it is a positive resistivity anomaly, with the gas level having approximately 5m depth. With the time-lapse analysis (Matlab script) between the obtained 2D resistivity sections from the site, it was possible to map how the biogas volume and position change in the landfill in relation to time. Our preliminary results show a preferential gas pathway through the subsurface studied area. A consistent relation between the gas depth and obtained microbiological data from archea and bacteria population was also observed.

  10. Impact of Time Lapse on ASCP Board of Certification Medical Laboratory Scientist (MLS) and Medical Laboratory Technician (MLT) Examination Scores.

    Science.gov (United States)

    Brown, Karen A; Fenn, JoAnn P; Freeman, Vicki S; Fisher, Patrick B; Genzen, Jonathan R; Goodyear, Nancy; Houston, Mary Lunz; O'Brien, Mary Elizabeth; Tanabe, Patricia A

    2015-01-01

    Research in several professional fields has demonstrated that delays (time lapse) in taking certification examinations may result in poorer performance by examinees. Thirteen states and/or territories require licensure for laboratory personnel. A core component of licensure is passing a certification exam. Also, many facilities in states that do not require licensure require certification for employment or preferentially hire certified individuals. To analyze examinee performance on the American Society for Clinical Pathology (ASCP) Board of Certification (BOC) Medical Laboratory Scientist (MLS) and Medical Laboratory Technician (MLT) certification examinations to determine whether delays in taking the examination from the time of program completion are associated with poorer performance. We obtained examination data from April 2013 through December 2014 to look for changes in mean (SD) exam scaled scores and overall pass/fail rates. First-time examinees (MLS: n = 6037; MLT, n = 3920) were divided into 3-month categories based on the interval of time between date of program completion and taking the certification exam. We observed significant decreases in mean (SD) scaled scores and pass rates after the first quarter in MLS and MLT examinations for applicants who delayed taking their examination until the second, third, and fourth quarter after completing their training programs. Those who take the ASCP BOC MLS and MLT examinations are encouraged to do so shortly after completion of their educational training programs. Delays in taking an exam are generally not beneficial to the examinee and result in poorer performance on the exam. Copyright© by the American Society for Clinical Pathology (ASCP).

  11. Time-lapse Mise-á-la-Masse measurements and modeling for tracer test monitoring in a shallow aquifer

    Science.gov (United States)

    Perri, Maria Teresa; De Vita, Pantaleone; Masciale, Rita; Portoghese, Ivan; Chirico, Giovanni Battista; Cassiani, Giorgio

    2018-06-01

    The main goal of this study is to evaluate the reliability of the Mise-á-la-Masse (MALM) technique associated with saline tracer tests for the characterization of groundwater flow direction and velocity. The experimental site is located in the upper part of the Alento River alluvial plain (Campania Region, Southern Italy). In this paper we present the hydrogeological setting, the experimental setup and the relevant field results. Subsequently, we compare those data against the simulated results obtained with a 3D resistivity model of the test area, coupled with a model describing the Advection - Dispersion equation for continuous tracer injection. In particular, we calculate a series of 3D forward solutions starting from a reference model, all derived from electrical tomography results, but taking into consideration different values of mean flow velocity and directions. Each electrical resistivity 3D model is used to produce synthetic voltage maps for MALM surveys. Finally, the synthetic MALM voltage maps are compared with the ones measured in the field in order to assess the information content of the MALM dataset with respect to the groundwater field characteristics. The results demonstrate that the information content of the MALM data is sufficient to define important characteristics of the aquifer geometry and properties. This work shows how a combination of three-dimensional time-lapse modeling of flow, tracer transport and electrical current can substantially contribute towards a quantitative interpretation of MALM measurements during a saline tracer test. This approach can thus revive the use of MALM as a practical, low cost field technique for tracer test monitoring and aquifer hydrodynamic characterization.

  12. Single-cell time-lapse analysis of depletion of the universally conserved essential protein YgjD

    Directory of Open Access Journals (Sweden)

    Ackermann Martin

    2011-05-01

    Full Text Available Abstract Background The essential Escherichia coli gene ygjD belongs to a universally conserved group of genes whose function has been the focus of a number of recent studies. Here, we put ygjD under control of an inducible promoter, and used time-lapse microscopy and single cell analysis to investigate the phenotypic consequences of the depletion of YgjD protein from growing cells. Results We show that loss of YgjD leads to a marked decrease in cell size and termination of cell division. The transition towards smaller size occurs in a controlled manner: cell elongation and cell division remain coupled, but cell size at division decreases. We also find evidence that depletion of YgjD leads to the synthesis of the intracellular signaling molecule (pppGpp, inducing a cellular reaction resembling the stringent response. Concomitant deletion of the relA and spoT genes - leading to a strain that is uncapable of synthesizing (pppGpp - abrogates the decrease in cell size, but does not prevent termination of cell division upon YgjD depletion. Conclusions Depletion of YgjD protein from growing cells leads to a decrease in cell size that is contingent on (pppGpp, and to a termination of cell division. The combination of single-cell timelapse microscopy and statistical analysis can give detailed insights into the phenotypic consequences of the loss of essential genes, and can thus serve as a new tool to study the function of essential genes.

  13. Pre-stack estimation of time-lapse seismic velocity changes : an example from the Sleipner CO2-sequestration project

    International Nuclear Information System (INIS)

    Ghaderi, A.; Landro, M.; Ghaderi, A.

    2005-01-01

    Carbon dioxide (CO 2 ) is being injected into a shallow sand formation at around a 1,000 metre depth at the Sleipner Field located in the North Sea. It is expected that the CO 2 injected in the bottom of the formation, will form a plume consisting of CO 2 accumulating in thin lenses during migration up through the reservoir. Several studies have been published using stacked seismic data from 1994, 1999, 2001 and 2002. A thorough analysis of post-stack seismic data from the Sleipner CO2-Sequestration Pilot Project was conducted. Interpretation of seismic data is usually done on post-stack data. For a given subsurface reflection point, seismic data are acquired for various incidence angles, typically 40 angles. These 40 seismic signals are stacked together in order to reduce noise. The term pre-stack refers to seismic data prior to this step. For hydrocarbon-related 4-dimensional seismic studies, travel time shift estimations have been used. This paper compared pre-stack and post-stack estimation of average velocity changes based on measured 4-dimensional travel time shifts. It is more practical to compare estimated velocity changes than the actual travel time changes, since the time shifts vary with offset for pre-stack time-lapse seismic analysis. It was concluded that the pre-stack method gives smaller velocity changes when estimated between two key horizons. Therefore, pre-stack travel time analysis in addition to conventional post-stack analysis is recommended. 6 refs., 12 figs

  14. Documentation of normal and leukemic myelopoietic progenitor cells with high-resolution phase-contrast time-lapse cinematography.

    Science.gov (United States)

    Boll, I T

    2001-08-01

    The high-resolution phase-contrast, time-lapse cinematography using oil immersion lenses and 16-mm film demonstrates the kinetic cell events as maturation, locomotion, mitosis, and apoptosis of cells cultivated at 37 degrees C for up to 10 days. 0.5 v/v frozen-thawed sera with presumably high cytokine concentrations were added to the plasma or agar clot. Vital progenitor cells from human bone marrow and blood have a large, bright, unstructured nucleus with a large nucleolus and a narrow rim of cytoplasm (nuclear/cytoplasmic volume ratio = 0.7). Their nuclei are 6-14 micrometer in diameter and double their volume within 8 h. Many (70%) move at a mean speed of 2 micrometer/min, and many (30%) multiply with alpha-2alpha mitoses, generating progenitor cell families. Various disturbances during the course of mitosis lead to the formation of polyploid cells, thereby yielding the megakaryocytic cell line. Some of the progenitor cells undergo asymmetric alpha-alphan mitoses: One of the two initially identical daughter cells remains a progenitor cell in the morphological sense, whereas the other daughter cell - depending on the size of its mother cell - matures in the same culture medium to form a granulocytopoietic, monocytopoietic or erythrocytopoietic cell line. - In acute myeloid leukemias (AML), the blasts and their nuclei are slightly larger than the corresponding progenitor cells and move faster (5 micrometer/min). Symmetric alpha-2alpha mitoses permit unlimited multiplication of the leukemic blasts if contact with cytotoxic lymphocytes does not render them apoptotic. This results in more stromal cells than normal. Granulocytopenia, monocytopenia, and anemia occur due to the genetic impairment of signaling control for asymmetric alpha-alphan mitoses, and thrombocytopenia occurs due to the reduction in polyploidization. Copyright 2001 S. Karger GmbH, Freiburg

  15. Proliferation of pulmonary endothelial cells: time-lapse cinematography of growth to confluence and restitution of monolayer after wounding.

    Science.gov (United States)

    Ryan, U S; Absher, M; Olazabal, B M; Brown, L M; Ryan, J W

    1982-01-01

    A fundamental characteristic of vascular endothelium is that it exists as a monolayer, a condition that must be met in both vascular growth and repair. Maintenance of the monolayer is important both for the exchange of nutrients and for interactions between blood solutes and endothelial enzymes and transport systems. We have used time-lapse cinematography to compare proliferative behavior of bovine pulmonary endothelial cells in (1) establishment of a monolayer from a low-density seed (7.5 X 10(4) cells in a 60 mm dish) and (2) restitution of a confluent monolayer (approx. 2.9 x 10(6) cells in a 60 mm dish) following a mechanical wound (removal of cells from an area 5 x 15 mm by scraping). Culture 2 was not refed after wounding. In culture 2, approx. 30% of the cells accounted for repopulation (confluence in 40 hr). In culture 1, all cells entered into division. Participating cells of culture 2 began division immediately (69 divisions/filmed area in 10 hr, vs. four divisions in culture 1). Interdivision times (IDT) were longer and relatively constant in culture 1 until near confluence; none were less than 10 h, whereas in 2, 24% of the IDT's were less than or equal to 10 hr. Remarkably, IDTs of culture 2 decreased steadily until confluence was re-established. Cell migration in culture 1 was multidirectional while direction of migration in culture 2 was always into the wound area. Mean migration rate (MIG) in culture 2 was related to the site of origin of the cells, those dividing farthest from the unwounded area had fastest MIGs. Neither culture formed more than a single layer of cells. Although the cell kinetics of cultures 1 and 2 differed, the same goal, confluence, was achieved in either case.

  16. Time lapse survey plan on the first offshore methane hydrate production test in 2013 around the eastern Nankai Trough area by multi-component OBC seismic tool

    Science.gov (United States)

    Inamori, T.; Hayashi, T.; Asakawa, E.; Takahashi, H.; Saeki, T.

    2011-12-01

    We are planning to conduct the multi-component ocean bottom cable (hereafter OBC) seismic survey to monitor the methane hydrate dissociation zone at the 1st offshore methane hydrate production test site in the eastern Nankai Trough, Japan, in 2013. We conducted the first OBC survey in the methane hydrate concentrated zone around the eastern Nankai Trough area in 2006 by RSCS which we developed. We obtained to the good image of methane hydrate bearing layer by P-P section as similar as the conventional surface seismic survey. However, we could not obtain the good image from P-S section compared with P-P section. On the other hand, we studied the sonic velocity distribution at the Mallik 2nd production test before and after in 2007, by the sonic tool data. We could clearly delineate the decrease of S-wave velocity, however, we could not detect the decrease of P-wave velocity because of the presence of the dissociated methane gas from methane hydrate. From these reason we guess the S-wave data is more proper to delineate the condition of the methane hydrate zone at the methane hydrate production tests than P-wave data. We are now developing the new OBC system, which we call Deep-sea Seismic System (hereafter DSS). The sensor of the DSS will install three accelerometers and one hydrophone. A feasibility study to detect the methane hydrate dissociation with the DSS was carried out and we found that the methane hydrate dissociation could be detected with the DSS depending on the zone of the dissociation. And the baseline survey will be held at the 1st offshore methane hydrate production test site in summer 2012. Two monitoring surveys are planned after the methane hydrate production test in 2013. We believe that we will get the good images to delineate the methane hydrate dissociated zone from this time lapse survey. The Authors would like to thank METI, MH21 consortium and JOGMEC for permissions to publish this paper.

  17. Deformed two-photon squeezed states in noncommutative space

    International Nuclear Information System (INIS)

    Zhang Jianzu

    2004-01-01

    Recent studies on nonperturbation aspects of noncommutative quantum mechanics explored a new type of boson commutation relations at the deformed level, described by deformed annihilation-creation operators in noncommutative space. This correlated boson commutator correlates different degrees of freedom, and shows an essential influence on dynamics. This Letter devotes to the development of formalism of deformed two-photon squeezed states in noncommutative space. General representations of deformed annihilation-creation operators and the consistency condition for the electromagnetic wave with a single mode of frequency in noncommunicative space are obtained. Two-photon squeezed states are studied. One finds that variances of the dimensionless Hermitian quadratures of the annihilation operator in one degree of freedom include variances in the other degree of freedom. Such correlations show the new feature of spatial noncommutativity and allow a deeper understanding of the correlated boson commutator

  18. Meson production in two-photon interactions at LHC energies

    Energy Technology Data Exchange (ETDEWEB)

    Da Silva, D. T.; Goncalves, V. P.; Sauter, W. K. [Instituto de Fisica e Matematica, Universidade Federal de Pelotas, Campus Universitario UFPel, CP 354, 96010-900, Capao do Leao-RS (Brazil)

    2013-03-25

    The LHC opens a new kinematical regime at high energy, where several questions related to the description of the high-energy regime of the Quantum Chromodynamics (QCD) remain without satisfactory answers. Some open questions are the search for non-q-bar q resonances, the determination of the spectrum of q-bar q states and the identification of states with anomalous {gamma}{gamma} couplings. A possible way to study these problems is the study of meson production in two-photon interactions. In this contribution we calculate the meson production in two-photon interactions at LHC energies considering proton - proton collisions and estimate the total cross section for the production of the mesons {pi}, a, f, {eta} and {chi}.

  19. Two-photon polarization Fourier spectroscopy of metastable atomic hydrogen

    International Nuclear Information System (INIS)

    Duncan, A.J.; Beyer, H.-J.; Kleinpoppen, H.; Sheikh, Z.A,; B-Z Univ., Multan

    1997-01-01

    A novel Fourier-transform spectroscopic method using two-photon polarization to determine the spectral distribution of the two photons emitted in the spontaneous decay of metastable atomic hydrogen is described. The method uses birefringent retardation plates and takes advantage of the subtle interplay between the spectral properties and the entangled polarization properties of the radiation emitted in the decay. Assuming the validity of the theoretical spectral distribution, it is shown that the experimental results agree well with theory. On the other hand, success in solving the inverse problem of determining the spectral distribution from the experimental results is limited by the small number of experimental points. However, making reasonable assumptions it is deduced that the observed spectrum is characterized by a broadband signal of width (0.43 ± 0.06) x 10 16 rad s -1 and centre angular frequency (0.77 ± 0.03) x 10 16 rad s -1 in good agreement with the predictions of 0.489 x 10 16 rad s -1 and 0.775 x 10 16 rad s -1 , respectively, obtained from the theoretical spectral distribution modified to take account of the absorption of the two-photon radiation in air. The values of 1.5 fs for the coherence time and 440 nm for the coherence length for single photons of the two-photon pair which are obtained from the measured bandwidth imply that, in the ideal case, these values are determined by the essentially zero lifetime of the virtual intermediate state of the decay process rather than the long lifetime of the metastable state which, it is suggested, determines the coherence time and coherence length appropriate to certain types of fourth-order interference experiments. (Author)

  20. Enhancement of two-photon photoluminescence and SERS for low-coverage gold films

    DEFF Research Database (Denmark)

    Novikov, Sergey M.; Beermann, Jonas; Frydendahl, Christian

    2016-01-01

    Electromagnetic field enhancement (FE) effects occurring in thin gold films 3-12-nm are investigated with two-photon photoluminescence (TPL) and Raman scanning optical microscopies. The samples are characterized using scanning electron microscopy images and linear optical spectroscopy. TPL images...... exhibit a strong increase in the level of TPL signals for films thicknesses 3-8-nm, near the percolation threshold. For some thicknesses, TPL measurements reveal super-cubic dependences on the incident power. We ascribe this feature to the occurrence of very strongly localized and enhanced electromagnetic...

  1. Novel migrating mouse neural crest cell assay system utilizing P0-Cre/EGFP fluorescent time-lapse imaging

    Directory of Open Access Journals (Sweden)

    Kawakami Minoru

    2011-11-01

    Full Text Available Abstract Background Neural crest cells (NCCs are embryonic, multipotent stem cells. Their long-range and precision-guided migration is one of their most striking characteristics. We previously reported that P0-Cre/CAG-CAT-lacZ double-transgenic mice showed significant lacZ expression in tissues derived from NCCs. Results In this study, by embedding a P0-Cre/CAG-CAT-EGFP embryo at E9.5 in collagen gel inside a culture glass slide, we were able to keep the embryo developing ex vivo for more than 24 hours; this development was with enough NCC fluorescent signal intensity to enable single-cell resolution analysis, with the accompanying NCC migration potential intact and with the appropriate NCC response to the extracellular signal maintained. By implantation of beads with absorbed platelet-derived growth factor-AA (PDGF-AA, we demonstrated that PDGF-AA acts as an NCC-attractant in embryos. We also performed assays with NCCs isolated from P0-Cre/CAG-CAT-EGFP embryos on culture plates. The neuromediator 5-hydroxytryptamine (5-HT has been known to regulate NCC migration. We newly demonstrated that dopamine, in addition to 5-HT, stimulated NCC migration in vitro. Two NCC populations, with different axial levels of origins, showed unique distribution patterns regarding migration velocity and different dose-response patterns to both 5-HT and dopamine. Conclusions Although avian species predominated over the other species in the NCC study, our novel system should enable us to use mice to assay many different aspects of NCCs in embryos or on culture plates, such as migration, division, differentiation, and apoptosis.

  2. Two-Photon Absorption in Organometallic Bromide Perovskites

    KAUST Repository

    Walters, Grant

    2015-07-21

    Organometallic trihalide perovskites are solution processed semiconductors that have made great strides in third generation thin film light harvesting and light emitting optoelectronic devices. Recently it has been demonstrated that large, high purity single crystals of these perovskites can be synthesized from the solution phase. These crystals’ large dimensions, clean bandgap, and solid-state order, have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW-1 at 800 nm, comparable to epitaxial single crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  3. Particle Production in Two-Photon Collisions at Belle

    International Nuclear Information System (INIS)

    Nakazawa, Hideyuki

    2010-01-01

    Experimental study of η η production in two-photon collisions: The differential cross section for the process γ γ → η η has been measured in the kinematic range above the η η threshold, 1.096 GeV -1 data sample collected with the Belle detector at the KEKB e + e - collider. In the W range 1.1-2.0 GeV/c 2 we perform an analysis of resonance amplitudes for various partial waves; at higher energy we extract the contributions of χ cJ charmonia and compare the energy and angular dependence of the cross section with the predictions of theoretical models. Observation of η c (2S) in six-prong final states produced in two-photon collisions: We report the observation of η c (2S), produced in two-photon collisions, and decaying to the six-prong final states 3(π + π - ), K + K - 2(π + π - ), and K 0 S K + ππ + π - (including the charge-conjugate state). This analysis is based on a large data sample accumulated by the Belle experiment at the KEKB asymmetric-energy electron-positron collider. This is the first observation of decay modes of the η c (2S) other than K 0 S K + π - . (author)

  4. Two-photon exchange in elastic electron-nucleon scattering

    International Nuclear Information System (INIS)

    Blunden, P.G.; Melnitchouk, W.; Tjon, J.A.

    2005-01-01

    A detailed study of two-photon exchange in unpolarized and polarized elastic electron-nucleon scattering is presented, taking particular account of nucleon finite size effects. Contributions from nucleon elastic intermediate states are found to have a strong angular dependence, which leads to a partial resolution of the discrepancy between the Rosenbluth and polarization transfer measurements of the proton electric to magnetic form factor ratio, G E /G M . The two-photon exchange contribution to the longitudinal polarization transfer P L is small, whereas the contribution to the transverse polarization transfer P T is enhanced at backward angles by several percent, increasing with Q 2 . This gives rise to a small, E /G M obtained from the polarization transfer ratio P T /P L at large Q 2 . We also compare the two-photon exchange effects with data on the ratio of e + p to e - p cross sections, which is predicted to be enhanced at backward angles. Finally, we evaluate the corrections to the form factors of the neutron and estimate the elastic intermediate state contribution to the 3 He form factors

  5. Two-Photon Absorption in Organometallic Bromide Perovskites

    KAUST Repository

    Walters, Grant; Sutherland, Brandon R; Hoogland, Sjoerd; Shi, Dong; Comin, Riccardo; Sellan, Daniel P.; Bakr, Osman; Sargent, Edward H.

    2015-01-01

    Organometallic trihalide perovskites are solution processed semiconductors that have made great strides in third generation thin film light harvesting and light emitting optoelectronic devices. Recently it has been demonstrated that large, high purity single crystals of these perovskites can be synthesized from the solution phase. These crystals’ large dimensions, clean bandgap, and solid-state order, have provided us with a suitable medium to observe and quantify two-photon absorption in perovskites. When CH3NH3PbBr3 single crystals are pumped with intense 800 nm light, we observe band-to-band photoluminescence at 572 nm, indicative of two-photon absorption. We report the nonlinear absorption coefficient of CH3NH3PbBr3 perovskites to be 8.6 cm GW-1 at 800 nm, comparable to epitaxial single crystal semiconductors of similar bandgap. We have leveraged this nonlinear process to electrically autocorrelate a 100 fs pulsed laser using a two-photon perovskite photodetector. This work demonstrates the viability of organometallic trihalide perovskites as a convenient and low-cost nonlinear absorber for applications in ultrafast photonics.

  6. Bayesian Markov-Chain-Monte-Carlo inversion of time-lapse crosshole GPR data to characterize the vadose zone at the Arrenaes Site, Denmark

    DEFF Research Database (Denmark)

    Scholer, Marie; Irving, James; Zibar, Majken Caroline Looms

    2012-01-01

    We examined to what extent time-lapse crosshole ground-penetrating radar traveltimes, measured during a forced infiltration experiment at the Arreneas field site in Denmark, could help to quantify vadose zone hydraulic properties and their corresponding uncertainties using a Bayesian Markov...... distributions compared with the corresponding priors, which in turn significantly improves knowledge of soil hydraulic properties. Overall, the results obtained clearly demonstrate the value of the information contained in time-lapse GPR data for characterizing vadose zone dynamics.......-chain-Monte-Carlo inversion approach with different priors. The ground-penetrating radar (GPR) geophysical method has the potential to provide valuable information on the hydraulic properties of the vadose zone because of its strong sensitivity to soil water content. In particular, recent evidence has suggested...

  7. Time-Lapse Monitoring of an Engineering Scaled Excavation at Federal District, Brazil by Passive Ambient NoiseInterferometry

    Science.gov (United States)

    Cárdenas-Soto, M., Sr.; Hussain, Y.; Martinez-Carvajal, H., Sr.; Martino, S., Sr.; Rocha, M., Sr.

    2016-12-01

    gave Rayleigh wave velocity changes (dv/v=-dt/t). These changes correlated well with initiation and propagation of fracture at the face of this normal slope. It is concluded that cost effective technique, PANI has a good potential for the monitoring of time lapse changes of evolving fractures.

  8. Monitoring snowmelt and solute transport at Oslo airport by combining time-lapse electrical resistivity, soil water sampling and tensiometer measurements

    Science.gov (United States)

    Bloem, E.; French, H. K.

    2013-12-01

    Monitoring contaminant transport at contaminated sites requires optimization of the configuration of a limited number of samplings points combined with heterogeneous flow and preferential flowpaths. Especially monitoring processes in the unsaturated zone is a major challenge due to the limited volume monitored by for example suction cups and their risk to clog in a highly active degradation zone. To make progress on soil contamination assessment and site characterization there is a strong need to integrate field-sale extensively instrumented tools, with non-invasive (geophysical) methods which provide spatially integrated measurements also in the unsaturated zone. Examples of sites that might require monitoring activities in the unsaturated zone are airports with winter frost where large quantities of de-icing chemicals are used each winter; salt and contaminant infiltration along roads; constructed infiltration systems for treatment of sewerage or landfill seepage. Electrical resistivity methods have proved to be useful as an indirect measurement of subsurface properties and processes at the field-scale. The non-uniqueness of the interpretation techniques can be reduced by constraining the inversion through the addition of independent geophysical measurements along the same profile. Or interpretation and understanding of geophysical images can be improved by the combination with classical measurements of soil physical properties, soil suction, contaminant concentration and temperatures. In our experiment, at the research field station at Gardermoen, Oslo airport, we applied a degradable de-icing chemical and an inactive tracer to the snow cover prior to snowmelt. To study the solute transport processes in the unsaturated zone time-lapse cross borehole electrical resistivity tomography (ERT) measurements were conducted at the same time as soil water samples were extracted at multiple depths with suction cups. Measurements of soil temperature, and soil tension were

  9. A low-cost landslide displacement activity assessment from time-lapse photogrammetry and rainfall data: Application to the Tessina landslide site

    Science.gov (United States)

    Gabrieli, F.; Corain, L.; Vettore, L.

    2016-09-01

    Acquiring useful and reliable displacement data from a complex landslide site is often a problem because of large, localized and scattered erosive processes and deformations; the inaccessibility of the site; the high cost of instrumentation and maintenance. However, these data are of fundamental importance not only to hazard assessments but also to understanding the processes at the basis of slope evolution. In this framework, time-lapse photogrammetry can represent a good compromise; the low accuracy is compensated for by the wide-ranging and dense spatial displacement information that can be obtained with inexpensive equipment. Nevertheless, when large displacement monitoring data sets become available, the problem becomes the choice of the most suitable statistical model to describe the probability of movement and adequately simplify the complexity of a scattered, intermittent, and spatially inhomogeneous displacement field. In this paper, an automated displacement detection method, which is based on the absolute image differences and digital correlations from a sequence of photos, was developed and applied to a photographic survey activity at the head of the Tessina landslide (northeastern Italy). The method allowed us to simplify and binarize the displacement field and to recognize the intermittent activity and the peculiar behaviours of different parts of the landslide, which were identified and classified by combining geomorphological and geological information. Moreover, for the first time, sliding correlations between these areas were quantitatively estimated using time-series-based binary logistic regression and the definition of a probability-based directed graph of displacement occurrence that connected the source zones to the lower depletion basin and the main collector channel. Using rainfall data, event-based logistic and Poisson regression models were applied to the upper zones of the landslide to estimate the probability of movement of each scarp

  10. Live cell linear dichroism imaging reveals extensive membrane ruffling within the docking structure of natural killer cell immune synapses

    DEFF Research Database (Denmark)

    Benninger, Richard K P; Vanherberghen, Bruno; Young, Stephen

    2009-01-01

    We have applied fluorescence imaging of two-photon linear dichroism to measure the subresolution organization of the cell membrane during formation of the activating (cytolytic) natural killer (NK) cell immune synapse (IS). This approach revealed that the NK cell plasma membrane is convoluted...... into ruffles at the periphery, but not in the center of a mature cytolytic NK cell IS. Time-lapse imaging showed that the membrane ruffles formed at the initial point of contact between NK cells and target cells and then spread radialy across the intercellular contact as the size of the IS increased, becoming...... absent from the center of the mature synapse. Understanding the role of such extensive membrane ruffling in the assembly of cytolytic synapses is an intriguing new goal....

  11. Application of Time-Lapse Seismic Monitoring for the Control and Optimization of CO2 Enhanced Oil Recovery Operations

    Energy Technology Data Exchange (ETDEWEB)

    Brian Toelle

    2008-11-30

    This project, 'Application of Time-Lapse Seismic Monitoring for the Control and Optimization of CO{sub 2} Enhanced Oil Recovery Operations', investigated the potential for monitoring CO{sub 2} floods in carbonate reservoirs through the use of standard p-wave seismic data. This primarily involved the use of 4D seismic (time lapse seismic) in an attempt to observe and map the movement of the injected CO{sub 2} through a carbonate reservoir. The differences between certain seismic attributes, such as amplitude, were used for this purpose. This technique has recently been shown to be effective in CO{sub 2} monitoring in Enhanced Oil Recovery (EOR) projects, such as Weyborne. This study was conducted in the Charlton 30/31 field in the northern Michigan Basin, which is a Silurian pinnacle reef that completed its primary production in 1997 and was scheduled for enhanced oil recovery using injected CO{sub 2}. Prior to injection an initial 'Base' 3D survey was obtained over the field and was then processed and interpreted. CO{sub 2} injection within the main portion of the reef was conducted intermittently during 13 months starting in August 2005. During this time, 29,000 tons of CO{sub 2} was injected into the Guelph formation, historically known as the Niagaran Brown formation. By September 2006, the reservoir pressure within the reef had risen to approximately 2000 lbs and oil and water production from the one producing well within the field had increased significantly. The determination of the reservoir's porosity distribution, a critical aspect of reservoir characterization and simulation, proved to be a significant portion of this project. In order to relate the differences observed between the seismic attributes seen on the multiple 3D seismic surveys and the actual location of the CO{sub 2}, a predictive reservoir simulation model was developed based on seismic attributes obtained from the base 3D seismic survey and available well data. This

  12. NIR-emitting molecular-based nanoparticles as new two-photon absorbing nanotools for single particle tracking

    Science.gov (United States)

    Daniel, J.; Godin, A. G.; Clermont, G.; Lounis, B.; Cognet, L.; Blanchard-Desce, M.

    2015-07-01

    In order to provide a green alternative to QDs for bioimaging purposes and aiming at designing bright nanoparticles combining both large one- and two-photon brightness, a bottom-up route based on the molecular engineering of dedicated red to NIR emitting dyes that spontaneously form fluorescent organic nanoparticles (FONs) has been implemented. These fully organic nanoparticles built from original quadrupolar dyes are prepared using a simple, expeditious and green protocol that yield very small molecular-based nanoparticles (radius ~ 7 nm) suspension in water showing a nice NIR emission (λem=710 nm). These FONs typically have absorption coefficient more than two orders larger than popular NIR-emitting dyes (such as Alexa Fluor 700, Cy5.5 ….) and much larger Stokes shift values (i.e. up to over 5500 cm-1). They also show very large two-photon absorption response in the 800-1050 nm region (up to about 106 GM) of major promise for two-photon excited fluorescence microscopy. Thanks to their brightness and enhanced photostability, these FONs could be imaged as isolated nanoparticles and tracked using wide-field imaging. As such, thanks to their size and composition (absence of heavy metals), they represent highly promising alternatives to NIR-emitting QDs for use in bioimaging and single particle tracking applications. Moreover, efficient FONs coating was achieved by using a polymeric additive built from a long hydrophobic (PPO) and a short hydrophilic (PEO) segment and having a cationic head group able to interact with the highly negative surface of FONs. This electrostatically-driven interaction promotes both photoluminescence and two-photon absorption enhancement leading to an increase of two-photon brightness of about one order of magnitude. This opens the way to wide-field single particle tracking under two-photon excitation

  13. Neural plasticity explored by correlative two-photon and electron/SPIM microscopy

    Science.gov (United States)

    Allegra Mascaro, A. L.; Silvestri, L.; Costantini, I.; Sacconi, L.; Maco, B.; Knott, G. W.; Pavone, F. S.

    2013-06-01

    Plasticity of the central nervous system is a complex process which involves the remodeling of neuronal processes and synaptic contacts. However, a single imaging technique can reveal only a small part of this complex machinery. To obtain a more complete view, complementary approaches should be combined. Two-photon fluorescence microscopy, combined with multi-photon laser nanosurgery, allow following the real-time dynamics of single neuronal processes in the cerebral cortex of living mice. The structural rearrangement elicited by this highly confined paradigm of injury can be imaged in vivo first, and then the same neuron could be retrieved ex-vivo and characterized in terms of ultrastructural features of the damaged neuronal branch by means of electron microscopy. Afterwards, we describe a method to integrate data from in vivo two-photon fluorescence imaging and ex vivo light sheet microscopy, based on the use of major blood vessels as reference chart. We show how the apical dendritic arbor of a single cortical pyramidal neuron imaged in living mice can be found in the large-scale brain reconstruction obtained with light sheet microscopy. Starting from its apical portion, the whole pyramidal neuron can then be segmented and located in the correct cortical layer. With the correlative approach presented here, researchers will be able to place in a three-dimensional anatomic context the neurons whose dynamics have been observed with high detail in vivo.

  14. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    Science.gov (United States)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  15. Inclusive $D*^{+-}$ Production in Two-Photon Collisions at LEP

    CERN Document Server

    Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery P.; Anselmo, F.; Arefev, A.; Azemoon, T.; Aziz, T.; Bagnaia, P.; Bajo, A.; Baksay, G.; Baksay, L.; Baldew, S.V.; Banerjee, S.; Banerjee, Sw.; Barczyk, A.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Echenard, B.; Eline, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Ewers, A.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hakobyan, R.S.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Krenz, W.; Kruger, A.; Kunin, A.; Ladron de Guevara, P.; Laktineh, I.; Landi, G.; Lebeau, M.; Lebedev, A.; Lebrun, P.; Lecomte, P.; Lecoq, P.; Le Coultre, P.; Le Goff, J.M.; Leiste, R.; Levtchenko, M.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Lubelsmeyer, K.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mangeol, D.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Niessen, T.; Nisati, A.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Palomares, C.; Pandoulas, D.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.O.; Prokofiev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, M.A.; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Sanders, M.P.; Schafer, C.; Schegelsky, V.; Schmidt-Kaerst, S.; Schmitz, D.; Schopper, H.; Schotanus, D.J.; Schwering, G.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Siedenburg, T.; Son, D.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wallraff, W.; Wang, X.L.; Wang, Z.M.; Weber, M.; Wienemann, P.; Wilkens, H.; Wynhoff, S.; Xia, L.; Xu, Z.Z.; Yamamoto, J.; Yang, B.Z.; Yang, C.G.; Yang, H.J.; Yang, M.; Yeh, S.C.; Zalite, A.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zilizi, G.; Zimmermann, B.; Zoller, M.

    2002-01-01

    Inclusive D^{*+-} production in two-photon collisions is studied with the L3 detector at LEP, using 683 pb^{-1} of data collected at centre-of-mass energies from 183 to 208 GeV. Differential cross sections are determined as functions of the transverse momentum and pseudorapidity of the D^{*+-} mesons in the kinematic region 1 GeV e^+e^-D^{*+-}X)$ in this kinematical region is measured and the sigma(e^+e^- ---> e^+e^- cc{bar}X) cross section is derived. The measurements are compared with next-to-leading order perturbative QCD calculations.

  16. New two-photon based nanoscopic modalities and optogenetics

    DEFF Research Database (Denmark)

    Glückstad, Jesper

    that can be equipped with optical trapping handles for convenient mechanical control using only optical forces [6]. These microstructures illustrated above can be effectively handled with simultaneous top- and side-view on our BioPhotonics Workstation to undertake six-degree-of-freedom optical actuation...... of two-photon polymerised microstructures equipped with features easily entering the submicron-regime. Aided by European collaborators who fabricated test structures with built-in waveguides for us, we were able to put the idea of optically steerable freestanding waveguides – coined: wave-guided optical...

  17. Two photon production of charm states - a charmonium calculation

    International Nuclear Information System (INIS)

    Suaya, R.

    1980-03-01

    The comparison of the nonrelativistic multichannel calculation with the double Rutherford process shows that duality is satisfied, in the sense that if one averages the actual cross sections for a wide enough range of W one gets the same result as for nonstrongly interacting point like quarks. It has been shown for the case of e + e - annihilation that duality in this sense follows from nonrelativistic potential models; and it has been verified that this proof can be extended to two-photon processes

  18. Two-photon exclusive processes in quantum chromodynamics

    International Nuclear Information System (INIS)

    Brodsky, S.J.

    1986-07-01

    QCD predictions for γγ annihilation into single mesons, meson pairs, and baryon pairs are reviewed. Two-photon exclusive processes provide the most sensitive and practical measure of the distribution amplitudes, and thus a critical confrontation between QCD and experiment. Both the angular distribution and virtual photon mass dependence of these amplitudes are sensitive to the shapes of the phi (chi, Q). Novel effects involving the production of qq anti q anti q states at threshold are also discussed, and a new method is presented for systematically incorporating higher-order QCD corrections in γγ reactions

  19. Two-photon exclusive processes in quantum chromodynamics

    Energy Technology Data Exchange (ETDEWEB)

    Brodsky, S.J.

    1986-07-01

    QCD predictions for ..gamma gamma.. annihilation into single mesons, meson pairs, and baryon pairs are reviewed. Two-photon exclusive processes provide the most sensitive and practical measure of the distribution amplitudes, and thus a critical confrontation between QCD and experiment. Both the angular distribution and virtual photon mass dependence of these amplitudes are sensitive to the shapes of the phi (chi, Q). Novel effects involving the production of qq anti q anti q states at threshold are also discussed, and a new method is presented for systematically incorporating higher-order QCD corrections in ..gamma gamma.. reactions.

  20. Direct and indirect two-photon processes in semiconductors

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1986-07-01

    The expressions describing direct and indirect two-photon absorption in crystals are given. They are valid both near and far from the energy gap. A perturbative approach through two different band models is adopted. The effects of the non-parabolicity and the degeneracy of the energy bands are considered. The numerical results are compared with the other theories and with a recent experimental data in Zn and AgCl. It is shown that the dominant transition mechanisms are of the allowed-allowed type near and far from the gap for both direct and indirect processes. (author)

  1. {Lambda}{bar {Lambda}} production in two-photon interactions

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, S.; Kubota, Y.; Lattery, M.; ONeill, J.J.; Patton, S.; Poling, R.; Riehle, T.; Savinov, V.; Smith, A. [University of Minnesota, Minneapolis, Minnesota 55455 (United States); Alam, M.S.; Athar, S.B.; Ling, Z.; Mahmood, A.H.; Severini, H.; Timm, S.; Wappler, F. [State University of New York at Albany, Albany, New York 12222 (United States); Anastassov, A.; Blinov, S.; Duboscq, J.E.; Fujino, D.; Fulton, R.; Gan, K.K.; Hart, T.; Honscheid, K.; Kagan, H.; Kass, R.; Lee, J.; Spencer, M.B.; Sung, M.; Undrus, A.; Wanke, R.; Wolf, A.; Zoeller, M.M. [Ohio State University, Columbus, Ohio 43210 (United States); Nemati, B.; Richichi, S.J.; Ross, W.R.; Skubic, P.; Wood, M. [University of Oklahoma, Norman, Oklahoma 73019 (United States); Bishai, M.; Fast, J.; Gerndt, E.; Hinson, J.W.; Menon, N.; Miller, D.H.; Shibata, E.I.; Shipsey, I.P.; Yurko, M. [Purdue University, West Lafayette, Indiana 47907 (United States); Gibbons, L.; Johnson, S.D.; Kwon, Y.; Roberts, S.; Thorndike, E.H. [University of Rochester, Rochester, New York 14627 (United States); Jessop, C.P.; Lingel, K.; Marsiske, H.; Perl, M.L.; Schaffner, S.F.; Ugolini, D.; Wang, R.; Zhou, X. [Stanford Linear Accelerator Center, Stanford University, Stanford, California 94309 (United States); Coan, T.E.; Fadeyev, V.; Korolkov, I.; Maravin, Y.; Narsky, I.; Shelkov, V.; Staeck, J.; Stroynowski, R.; Volobouev, I.; Ye, J. [Southern Methodist University, Dallas, Texas 75275 (United States); Artuso, M.; Efimov, A.; Frasconi, F.; Gao, M.; Goldberg, M.; He, D.; Kopp, S.; Moneti, G.C.; Mountain, R.; Mukhin, Y.; Schuh, S.; Skwarnicki, T.; Stone, S.; Viehhauser, G.; Xing, X. [Syracuse University, Syracuse, New York 13244 (United States); Bartelt, J.; Csorna, S.E.; Jain, V.; Marka, S. [Vanderbilt University, Nashville, Tennessee 37235 (United States); Freyberger, A.; Gibaut, D.; Godang, R.; Kinoshita, K.; Lai, I.C.; Pomianowski, P.; Schrenk, S.; and others

    1997-09-01

    Using the CLEO detector at the Cornell e{sup +}e{sup {minus}} storage ring CESR we study the two-photon production of {Lambda}{bar {Lambda}}, making the first observation of {gamma}{gamma}{r_arrow}{Lambda}{bar {Lambda}}. We present the cross section for {gamma}{gamma}{r_arrow}{Lambda}{bar {Lambda}} as a function of the {gamma}{gamma} center of mass energy and compare it to that predicted by the quark-diquark model. {copyright} {ital 1997} {ital The American Physical Society}

  2. Quantitative monitoring of CO2 injection at Sleipner using seismic full waveform inversion in the time lapse mode and rock physics modeling

    International Nuclear Information System (INIS)

    Queisser, M.P.

    2012-01-01

    Carbon capture and sequestration is a technology to achieve a considerable deceleration of CO 2 emission promptly. Since 1996 one of the largest CO 2 storage projects is taking place at Sleipner in the Norwegian North Sea. In order to monitor injected CO 2 , time lapse seismic monitoring surveys have been carried out. Estimating subsurface parameters from the Sleipner seismic data is a challenging problem due to the specific geology of the storage reservoir, which is further complicated by injected CO 2 . Most seismic imaging methods enable only qualitative insights into the subsurface. Motivated by the need for a quantitative seismic monitoring of the injected CO 2 , I have applied 2D seismic full waveform inversion to seismic data sets from Sleipner from 1994 (baseline), 1999 and 2006 along three seismic lines to infer subsurface parameters and parameter changes in the storage reservoir. The P-wave velocity is the major parameter, as it is the most sensitive to CO 2 injection. An energy preconditioning of the gradient has been implemented. The usual source wavelet calibration did not prove to be reliable. An alternative source calibration has been successfully applied. By comparing seismic images with inversion results, I found that using seismic images to locate CO 2 accumulations in the subsurface may be misleading. The quantitative imaging approach using full waveform inversion resulted in a consistent evolution of the model parameter with time. Major reductions in P-wave velocity and hence the CO 2 accumulations could be quantitatively imaged down to a resolution of 10 m. Observed travel time shifts due to CO 2 injection are comparable to those derived from the inversion result. In order to estimate CO 2 saturations, rock physical concepts have been combined and extended to arrive at a rock physical formulation of the subsurface at Sleipner. I used pseudo Monte Carlo rock physics modeling to assess the influence of lithologic heterogeneity on the CO 2

  3. Ultra-large field-of-view two-photon microscopy

    OpenAIRE

    Tsai, Philbert S.; Mateo, Celine; Field, Jeffrey J.; Schaffer, Chris B.; Anderson, Matthew E.; Kleinfeld, David

    2015-01-01

    We present a two-photon microscope that images the full extent of murine cortex with an objective-limited spatial resolution across an 8 mm by 10 mm field. The lateral resolution is approximately 1 µm and the maximum scan speed is 5 mm/ms. The scan pathway employs large diameter compound lenses to minimize aberrations and performs near theoretical limits. We demonstrate the special utility of the microscope by recording resting-state vasomotion across both hemispheres of the murine brain thro...

  4. Nonlinear behavior of photoluminescence from silicon particles under two-photon excitation

    International Nuclear Information System (INIS)

    Xu Xingsheng; Yokoyama, Shiyoshi

    2011-01-01

    Two-photon excited fluorescence (TPEF) under continuous-wave excitation from silicon particles produced by a pulsed laser is investigated. Spectra and images of TPEF from silicon particles are studied under different excitation intensities and operation modes (continuous wave or pulse). It is found that the photoluminescence depends superlinearly on the excitation intensity and that the spectral shape and peaks vary with different silicon particles. The above phenomena show the nonlinear behavior of TPEF from silicon particles, and stimulated emission is a possible process.

  5. Two-photon mapping of localized field enhancements in thin nanostrip antennas

    DEFF Research Database (Denmark)

    Beermann, I.; Novikov, S.M.; Søndergaard, Thomas

    2008-01-01

    scanning optical microscopy, in which two-photon-excited photoluminescence (TPL) excited with a strongly focused laser beam at the wavelength 745 nm is detected. We use TPL images to map the local field enhancements from individual nanostrips at a resolution of 0.35µm and compare results with theoretical......Resonant scattering and local field enhancements by 11-nm-thin gold nanostrip antennas due to constructive interference of counter propagating slow surface plasmon polaritons is investigated. We characterize nanostrips of widths between 50-530 nm using both reflection spectroscopy and nonlinear...

  6. Single-organelle tracking by two-photon conversion

    Science.gov (United States)

    Watanabe, Wataru; Shimada, Tomoko; Matsunaga, Sachihiro; Kurihara, Daisuke; Fukui, Kiichi; Shin-Ichi Arimura, Shin-Ichi; Tsutsumi, Nobuhiro; Isobe, Keisuke; Itoh, Kazuyoshi

    2007-03-01

    Spatial and temporal information about intracellular objects and their dynamics within a living cell are essential for dynamic analysis of such objects in cell biology. A specific intracellular object can be discriminated by photoactivatable fluorescent proteins that exhibit pronounced light-induced spectral changes. Here, we report on selective labeling and tracking of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. We performed selective labeling of a single mitochondrion in a living tobacco BY-2 cell using two-photon photoconversion of Kaede. Using this technique, we demonstrated that, in plants, the directed movement of individual mitochondria along the cytoskeletons was mediated by actin filaments, whereas microtubules were not required for the movement of mitochondria. This single-organelle labeling technique enabled us to track the dynamics of a single organelle, revealing the mechanisms involved in organelle dynamics. The technique has potential application in direct tracking of selective cellular and intracellular structures.

  7. Inclusive D*(+/-) production in two photon collisions at LEP

    CERN Document Server

    Prokofiev, Denis Olegovich

    2001-01-01

    In this thesis I present my results on the measurement of the open charm production in two-photon collision events done with the L3 detector at Large Electron Positron machine (LEP). The data sample was collected from 1997 through 2000 at center-of-mass energies ranging from 183 GeV to 209 GeV, corresponding to a total integrated luminosity of 683.4pb −1. The open charm production in two-photon collision events extrapolated to the full phase space is estimated to be: s&parl0;e+e-&rarrr;e +e-cc&d1;X&parr0;=9 23±69±109±222pb. The differential cross sections d s /dpT(D*±) and d s /d:η(D*±): are also measured as functions of transverse momentum pT(D*±) and the absolute value of pseudorapidity :η(D*±):, respectively. A fit to the data estimating the relative contributions of Direct and Resolved open charm production mechanisms is performed, giving (28.7 ± 5.6)% and (71.3 ± 8.8)%, respectively. Using those relative fractions, the Direct and Resolved process cross sections yield: s&p...

  8. Temperature dependence of the two photon absorption in indium arsenide

    International Nuclear Information System (INIS)

    Berryman, K.W.; Rella, C.W.

    1995-01-01

    Nonlinear optical processes in semiconductors have long been a source of interesting physics. Two photon absorption (TPA) is one such process, in which two photons provide the energy for the creation of an electron-hole pair. Researchers at other FEL centers have studied room temperature TPA in InSb, InAs, and HgCdTe. Working at the Stanford Picosecond FEL Center, we have extended and refined this work by measuring the temperature dependence of the TPA coefficient in InAs over the range from 80 to 350 K at four wavelengths: 4.5, 5.06, 6.01, and 6.3 microns. The measurements validate the functional dependence of recent band structure calculations with enough precision to discriminate parabolic from non-parabolic models, and to begin to observe smaller effects, such as contributions due to the split-off band. These experiments therefore serve as a strong independent test of the Kane band theory, as well as providing a starting point for detailed observations of other nonlinear absorption mechanisms

  9. Two-photon emission and multiphoton absorption by atoms

    International Nuclear Information System (INIS)

    Mu, X.

    1988-01-01

    This thesis consists of investigations of two problems concerning photon-atom interactions. The first topic deals with two-photon transitions in atomic inner shells. An independent-particle model has been used to describe the two-photon transitions between different inner-shell electron states. The first relativistic self-consistent-field calculation of these transition rates in Ag, Mo, and Xe has been carried out. The theoretical results are compared with recent measurements. Good agreement with measured rates is found except in some cases where more reliable experiments still need to be done. The second topic is multiphoton multiionization of atoms. The maximum entropy principle has been employed in this theoretical investigation. A detailed statistical analysis of measured ionic charge distributions produced in strong laser pulses has been carried out. The results of this analysis indicates that the charge-state distribution is a Poissonian, rather than the binomial which prevails under infrared radiation, and hence that ionization occurs stepwise during the pulse. This result is shown to be consistent with experimental data

  10. Monte Carlo simulation of two-photon processes

    International Nuclear Information System (INIS)

    Daverveldt, P.H.W.M.

    1985-01-01

    During the last two decades e + e - collider experiments provided physicists with a wealth of important discoveries concerning elementary particle physics. This thesis explains in detail how the Monte Carlo approach can be applied to establish the comparison between two-photon experiments and theory. The author describes the main motives for and objectives of two-photon research. He defines the kinematics and pays attention to some special kinematical regions. Also a popular approximation for the exact differential cross section is reviewed. Next he discusses the calculation of the complete lowest order cross section for processes with four leptons in the final state and for reactions such as e + e - →e + e - qanti q, e + e - →μ + μ - qanti q. Radiative corrections to the multiperipheral diagrams are considered. The author explains in detail the distinction between soft and hard photon corrections which turns out to be somewhat more tricky than in the case of radiative corrections to one-photon processes. Finally, he presents some results which were obtained by using the event generators. (Auth.)

  11. Quantum theory of two-photon wavepacket interference in a beamsplitter

    International Nuclear Information System (INIS)

    Wang, Kaige

    2006-01-01

    A general theory is derived for the interference of a two-photon wavepacket in a beamsplitter. The theory is presented in the Schroedinger picture so that the quantum nature of the two-photon interference is explicitly revealed. We find that the topological symmetry of the probability-amplitude spectrum of the two-photon wavepacket dominates the nature of the two-photon interference, which may be distinguished by the increase or decrease of the coincidence probability in the absence of interference. However, two-photon entanglement can be identified by the nature of the interference. We demonstrate the necessary and sufficient conditions for perfect two-photon interference. It is shown that a two-photon entangled state with an anti-symmetric spectrum passes through a 50/50 beamsplitter with perfect transparency. The theory provides us with a unified understanding of the various two-photon interference effects. (topical review)

  12. Estimation of soil hydraulic parameters in the field by integrated hydrogeophysical inversion of time-lapse ground-penetrating radar data

    KAUST Repository

    Jadoon, Khan

    2012-01-01

    An integrated hydrogeophysical inversion approach was used to remotely infer the unsaturated soil hydraulic parameters from time-lapse ground-penetrating radar (GPR) data collected at a fixed location over a bare agricultural field. The GPR model combines a full-waveform solution of Maxwell\\'s equations for three-dimensional wave propaga- tion in planar layered media together with global reflection and transmission functions to account for the antenna and its interactions with the medium. The hydrological simu- lator HYDRUS-1D was used with a two layer single- and dual-porosity model. The radar model was coupled to the hydrodynamic model, such that the soil electrical properties (permitivity and conductivity) that serve as input to the GPR model become a function of the hydrodynamic model output (water content), thereby permiting estimation of the soil hydraulic parameters from the GPR data in an inversion loop. To monitor the soil water con- tent dynamics, time-lapse GPR and time domain reflectometry (TDR) measurements were performed, whereby only GPR data was used in the inversion. Significant effects of water dynamics were observed in the time-lapse GPR data and in particular precipitation and evaporation events were clearly visible. The dual porosity model provided betier results compared to the single porosity model for describing the soil water dynamics, which is sup- ported by field observations of macropores. Furthermore, the GPR-derived water content profiles reconstructed from the integrated hydrogeophysical inversion were in good agree- ment with TDR observations. These results suggest that the proposed method is promising for non-invasive characterization of the shallow subsurface hydraulic properties and moni- toring water dynamics at the field scale. © Soil Science Society of America.

  13. Two-photon Absorption In Quantum Dots,quantum Dashes And Related Materials

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Ravinder

    2009-08-31

    We have proposed the use of USQDs for various deep-tissue biological imaging applications, notably wavelength-multiplexed multicolor imaging and intra-nuclear studies such as those involving cell apoptosis, and have studied the issue of maximizing two-photon absorption-induced fluorescence (TPAF) signals from CdSe/ZnS USQDs to be used for this application. In particular, using 2 nm USQDs, we have shown that the TPAF signal at 780 nm is ~ 8 times that at 850 nm and 68 times that at 900 nm, two wavelengths that have been used in previous studies using CdSe/ZnS SQDs for deep-tissue imaging of biological studies via TPAF .

  14. Wide-band acousto-optic deflectors for large field of view two-photon microscope.

    Science.gov (United States)

    Jiang, Runhua; Zhou, Zhenqiao; Lv, Xiaohua; Zeng, Shaoqun

    2012-04-01

    Acousto-optic deflector (AOD) is an attractive scanner for two-photon microscopy because it can provide fast and versatile laser scanning and does not involve any mechanical movements. However, due to the small scan range of available AOD, the field of view (FOV) of the AOD-based microscope is typically smaller than that of the conventional galvanometer-based microscope. Here, we developed a novel wide-band AOD to enlarge the scan angle. Considering the maximum acceptable acoustic attenuation in the acousto-optic crystal, relatively lower operating frequencies and moderate aperture were adopted. The custom AOD was able to provide 60 MHz 3-dB bandwidth and 80% peak diffraction efficiency at 840 nm wavelength. Based on a pair of such AOD, a large FOV two-photon microscope was built with a FOV up to 418.5 μm (40× objective). The spatiotemporal dispersion was compensated simultaneously with a single custom-made prism. By means of dynamic power modulation, the variation of laser intensity within the FOV was reduced below 5%. The lateral and axial resolution of the system were 0.58-2.12 μm and 2.17-3.07 μm, respectively. Pollen grain images acquired by this system were presented to demonstrate the imaging capability at different positions across the entire FOV. © 2012 American Institute of Physics

  15. Two-photon-based photoactivation in live zebrafish embryos.

    Science.gov (United States)

    Russek-Blum, Niva; Nabel-Rosen, Helit; Levkowitz, Gil

    2010-12-24

    Photoactivation of target compounds in a living organism has proven a valuable approach to investigate various biological processes such as embryonic development, cellular signaling and adult physiology. In this respect, the use of multi-photon microscopy enables quantitative photoactivation of a given light responsive agent in deep tissues at a single cell resolution. As zebrafish embryos are optically transparent, their development can be monitored in vivo. These traits make the zebrafish a perfect model organism for controlling the activity of a variety of chemical agents and proteins by focused light. Here we describe the use of two-photon microscopy to induce the activation of chemically caged fluorescein, which in turn allows us to follow cell's destiny in live zebrafish embryos. We use embryos expressing a live genetic landmark (GFP) to locate and precisely target any cells of interest. This procedure can be similarly used for precise light induced activation of proteins, hormones, small molecules and other caged compounds.

  16. Two-photon resonant, stimulated processes in krypton and xenon

    International Nuclear Information System (INIS)

    Miller, J.C.

    1988-11-01

    Both on-axis and conical emissions have been observed following two-photon pumping of the 5p states of krypton and the 6p', 7p, 8p, and 4f states of xenon. In the former case, coherent emissions from the 5p states to the 5s are observed, and in the latter case, many p→s, d→p, and f→d cascade emissions are observed. By analogy to the well-studied alkali and alkaline earth examples, the emissions are discussed in terms of amplified spontaneous emission (ASE), stimulated hyper-Raman scattering, and parametric four-wave mixing. The physical processes responsible for the conical emission and for intensity anomalies in the xenon p→s emissions are not understood at present. Interference effects due to coherent cancellation between competing excitation pathways may be occurring. 4 refs., 3 figs

  17. N-dimensional integrability from two-photon coalgebra symmetry

    International Nuclear Information System (INIS)

    Ballesteros, Angel; Blasco, Alfonso; Herranz, Francisco J

    2009-01-01

    A wide class of Hamiltonian systems with N degrees of freedom and endowed with, at least, (N - 2) functionally independent integrals of motion in involution is constructed by making use of the two-photon Lie-Poisson coalgebra (h 6 , Δ). The set of (N - 2) constants of the motion is shown to be a universal one for all these Hamiltonians, irrespective of the dependence of the latter on several arbitrary functions and N free parameters. Within this large class of quasi-integrable N-dimensional Hamiltonians, new families of completely integrable systems are identified by finding explicitly a new independent integral I through the analysis of the sub-coalgebra structure of h 6 . In particular, new completely integrable N-dimensional Hamiltonians describing natural systems, geodesic flows and static electromagnetic Hamiltonians are presented

  18. Phonon-assisted two-photon exciton transitions in semiconductors

    International Nuclear Information System (INIS)

    Hassan, A.R.

    1987-08-01

    The theory of phonon-assisted two-photon transitions to excitonic states in semiconductors has been theoretically investigated. The effects of both the nonparabolicity of the band and the degeneracy of the valence band have been taken into account. Expressions for the absorption coefficient through different band models are calculated. The numerical applications to CdI 2 and GaP show that the 4-band model gives the dominant contribution which leads to a final s-exciton state. An exciton peak appears at an energy which is close to that recently observed in CdI 2 . The non-parabolic effect enhances the absorption coefficient by a two-order of magnitude. (author). 6 refs, 1 fig., 1 tab

  19. Brain refractive index measured in vivo with high-NA defocus-corrected full-field OCT and consequences for two-photon microscopy.

    Science.gov (United States)

    Binding, Jonas; Ben Arous, Juliette; Léger, Jean-François; Gigan, Sylvain; Boccara, Claude; Bourdieu, Laurent

    2011-03-14

    Two-photon laser scanning microscopy (2PLSM) is an important tool for in vivo tissue imaging with sub-cellular resolution, but the penetration depth of current systems is potentially limited by sample-induced optical aberrations. To quantify these, we measured the refractive index n' in the somatosensory cortex of 7 rats in vivo using defocus optimization in full-field optical coherence tomography (ff-OCT). We found n' to be independent of imaging depth or rat age. From these measurements, we calculated that two-photon imaging beyond 200 µm into the cortex is limited by spherical aberration, indicating that adaptive optics will improve imaging depth.

  20. Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue

    Science.gov (United States)

    Sakadžić, Sava; Roussakis, Emmanuel; Yaseen, Mohammad A.; Mandeville, Emiri T.; Srinivasan, Vivek J.; Arai, Ken; Ruvinskaya, Svetlana; Devor, Anna; Lo, Eng H.; Vinogradov, Sergei A.; Boas, David A.

    2010-01-01

    The ability to measure oxygen partial pressure (pO2) with high temporal and spatial resolution in three dimensions is crucial for understanding oxygen delivery and consumption in normal and diseased brain. Among existing pO2 measurement methods, phosphorescence quenching is optimally suited for the task. However, previous attempts to couple phosphorescence with two-photon laser scanning microscopy have faced substantial difficulties because of extremely low two-photon absorption cross-sections of conventional phosphorescent probes. Here, we report the first practical in vivo two-photon high-resolution pO2 measurements in small rodents’ cortical microvasculature and tissue, made possible by combining an optimized imaging system with a two-photon-enhanced phosphorescent nanoprobe. The method features a measurement depth of up to 250 µm, sub-second temporal resolution and requires low probe concentration. Most importantly, the properties of the probe allowed for the first direct high-resolution measurement of cortical extravascular (tissue) pO2, opening numerous possibilities for functional metabolic brain studies. PMID:20693997

  1. Porous Porphyrin-Based Organosilica Nanoparticles for NIR Two-Photon Photodynamic Therapy and Gene Delivery in Zebrafish

    KAUST Repository

    Mauriello Jimenez, Chiara

    2018-03-30

    Periodic mesoporous organosilica nanoparticles emerge as promising vectors for nanomedicine applications. Their properties are very different from those of well-known mesoporous silica nanoparticles as there is no silica source for their synthesis. So far, they have only been synthesized from small bis-silylated organic precursors. However, no studies employing large stimuli-responsive precursors have been reported on such hybrid systems yet. Here, the synthesis of porphyrin-based organosilica nanoparticles from a large octasilylated metalated porphyrin precursor is described for applications in near-infrared two-photon-triggered spatiotemporal theranostics. The nanoparticles display unique interconnected large cavities of 10-80 nm. The framework of the nanoparticles is constituted with J-aggregates of porphyrins, which endows them with two-photon sensitivity. The nanoparticle efficiency for intracellular tracking is first demonstrated by the in vitro near-infrared imaging of breast cancer cells. After functionalization of the nanoparticles with aminopropyltriethoxysilane, two-photon-excited photodynamic therapy in zebrafish is successfully achieved. Two-photon photochemical internalization in cancer cells of the nanoparticles loaded with siRNA is also performed for the first time. Furthermore, siRNA targeting green fluorescent protein complexed with the nanoparticles is delivered in vivo in zebrafish embryos, which demonstrates the versatility of the nanovectors for biomedical applications.

  2. Two-photon high-resolution measurement of partial pressure of oxygen in cerebral vasculature and tissue.

    Science.gov (United States)

    Sakadzić, Sava; Roussakis, Emmanuel; Yaseen, Mohammad A; Mandeville, Emiri T; Srinivasan, Vivek J; Arai, Ken; Ruvinskaya, Svetlana; Devor, Anna; Lo, Eng H; Vinogradov, Sergei A; Boas, David A

    2010-09-01

    Measurements of oxygen partial pressure (pO(2)) with high temporal and spatial resolution in three dimensions is crucial for understanding oxygen delivery and consumption in normal and diseased brain. Among existing pO(2) measurement methods, phosphorescence quenching is optimally suited for the task. However, previous attempts to couple phosphorescence with two-photon laser scanning microscopy have faced substantial difficulties because of extremely low two-photon absorption cross-sections of conventional phosphorescent probes. Here we report to our knowledge the first practical in vivo two-photon high-resolution pO(2) measurements in small rodents' cortical microvasculature and tissue, made possible by combining an optimized imaging system with a two-photon-enhanced phosphorescent nanoprobe. The method features a measurement depth of up to 250 microm, sub-second temporal resolution and requires low probe concentration. The properties of the probe allowed for direct high-resolution measurement of cortical extravascular (tissue) pO(2), opening many possibilities for functional metabolic brain studies.

  3. Assessment of human embryo development using morphological criteria in an era of time-lapse, algorithms and 'OMICS': is looking good still important?

    Science.gov (United States)

    Gardner, David K; Balaban, Basak

    2016-10-01

    With the worldwide move towards single embryo transfer there has been a renewed focus on the requirement for reliable means of assessing embryo viability. In an era of 'OMICS' technologies, and algorithms created through the use of time-lapse microscopy, the actual appearance of the human embryo as it progresses through each successive developmental stage to the blastocyst appears to have been somewhat neglected in recent years. Here we review the key features of the human preimplantation embryo and consider the relationship between morphological characteristics and developmental potential. Further, the impact of the culture environment on morphological traits, how key morphological qualities reflect aspects of embryo physiology, and how computer-assisted analysis of embryo morphology may facilitate a more quantitative approach to selection are discussed. The clinical introduction of time-lapse systems has reopened our eyes and given us a new vantage point from which to view the beauty of the initial stages of human life. Rather than a future in which the morphology of the embryo is deemed irrelevant, we propose that key features, such as multinucleation, cell size and blastocyst differentiation should be included in future iterations of selection/deselection algorithms. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.For Permissions, please email: journals.permissions@oup.com.

  4. Targeted two-photon photodynamic therapy for the treatment of subcutaneous tumors

    Science.gov (United States)

    Spangler, Charles W.; Starkey, Jean R.; Meng, Fanqing; Gong, Aijun; Drobizhev, Mikhail; Rebane, Aleksander; Moss, B.

    2005-04-01

    Photodynamic therapy (PDT) has developed into a mature technology over the past several years, and is currently being exploited for the treatment of a variety of cancerous tumors, and more recently for age-related wet macular degeneration of the eye. However, there are still some unresolved problems with PDT that are retarding a more general acceptance in clinical settings, and thus, for the most part, the treatment of most cancerous rumors still involves some combination of invasive surgery, chemotherapy and radiation treatment, particularly subcutaneous tumors. Currently approved PDT agents are activated in the Visible portion of the spectrum below 700 nm, Laser light in this spectral region cannot penetrate the skin more than a few millimeters, and it would be more desirable if PDT could be initiated deep in the Near-infrared (NIR) in the tissue transparency window (700-1000 nm). MPA Technologies, Inc. and Rasiris, Inc. have been co-developing new porphyrin PDT designed to have greatly enhanced intrinsic two-photon cross-sections (>800 GM units) whose two-photon absorption maxima lie deep in the tissue transparency window (ca. 780-850 nm), and have solubility characteristics that would allow for direct IV injection into animal models. Classical PDT also suffers from the lengthy time necessary for accumulation at the tumor site, a relative lack of discrimination between healthy and diseased tissue, particularly at the tumor margins, and difficulty in clearing from the system in a reasonable amount of time post-PDT. We have recently discovered a new design paradigm for the delivery of our two-photon activated PDT agents by incorporating the porphyrins into a triad ensemble that includes a small molecule targeting agent that directs the triad to over-expressed tumor receptor sites, and a NIR one-photon imaging agent that allows the tracking of the triad in terms of accumulation and clearance rates. We are currently using these new two-photon PDT triads in efficacy

  5. Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe

    International Nuclear Information System (INIS)

    Chen, Qian; Yang, Jinfeng; Li, Yinhui; Zheng, Jing; Yang, Ronghua

    2015-01-01

    Development of efficient methods for detection of endogenous H 2 S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H 2 S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H 2 S imaging have been designed, but real-time imaging of endogenous H 2 S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H 2 S) for highly sensitive and fast monitoring and imaging H 2 S levels in living cells and tissues. In the presence of H 2 S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H 2 S probes. With two-photon excitation, TPP-H 2 S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H 2 S is applied for fast imaging of H 2 S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H 2 S level in different viscera by vivisection and found that the distribution of endogenous H 2 S mostly in brain, liver and lung. The excellent sensing properties of TPP-H 2 S make it a practically useful tool for further studying biological roles of H 2 S. - Highlights: • This two-photon probe exhibits an improved sensitivity and response time to H 2 S. • This probe shows excellent membrane permeability and fast visualization of H 2 S in living cells and tissues. • This probe is successfully applied to measure the endogenously produced H 2 S levels in different viscera of mouse.

  6. Visualizing and quantifying movement from pre-recorded videos: The spectral time-lapse (STL algorithm [v1; ref status: indexed, http://f1000r.es/2qo

    Directory of Open Access Journals (Sweden)

    Christopher R Madan

    2014-01-01

    Full Text Available When studying animal behaviour within an open environment, movement-related data are often important for behavioural analyses. Therefore, simple and efficient techniques are needed to present and analyze the data of such movements. However, it is challenging to present both spatial and temporal information of movements within a two-dimensional image representation. To address this challenge, we developed the spectral time-lapse (STL algorithm that re-codes an animal’s position at every time point with a time-specific color, and overlays it with a reference frame of the video, to produce a summary image. We additionally incorporated automated motion tracking, such that the animal’s position can be extracted and summary statistics such as path length and duration can be calculated, as well as instantaneous velocity and acceleration. Here we describe the STL algorithm and offer a freely available MATLAB toolbox that implements the algorithm and allows for a large degree of end-user control and flexibility.

  7. Two-Photon Absorbing Molecules as Potential Materials for 3D Optical Memory

    Directory of Open Access Journals (Sweden)

    Kazuya Ogawa

    2014-01-01

    Full Text Available In this review, recent advances in two-photon absorbing photochromic molecules, as potential materials for 3D optical memory, are presented. The investigations introduced in this review indicate that 3D data storage processing at the molecular level is possible. As 3D memory using two-photon absorption allows advantages over existing systems, the use of two-photon absorbing photochromic molecules is preferable. Although there are some photochromic molecules with good properties for memory, in most cases, the two-photon absorption efficiency is not high. Photochromic molecules with high two-photon absorption efficiency are desired. Recently, molecules having much larger two-photon absorption cross sections over 10,000 GM (GM= 10−50 cm4 s molecule−1 photon−1 have been discovered and are expected to open the way to realize two-photon absorption 3D data storage.

  8. Microspot two-photon photoemission spectroscopy for CuPc film on HOPG

    International Nuclear Information System (INIS)

    Yamada, T.; Yamamoto, R.; Munakata, T.

    2015-01-01

    Highlights: • Unoccupied levels of CuPc/HOPG are assigned by using 2PPE microspectroscopy. • Lateral distribution of unoccupied energy levels is imaged. • Modified IPS stabilized by the hole localized in the 2nd layer molecule is identified. - Abstract: Microspot two-photon photoemission (micro-2PPE) spectroscopy has been applied to measure the lateral distribution of unoccupied levels on copper phthalocyanine (CuPc) film on HOPG. In addition to the LUMO-derived level and the image potential state (IPS) on the film, we identified the modified IPS which is stabilized by the hole localized in a molecule. We show that modified IPS is observed only on bilayer area, reflecting the localization of the hole in a molecule. The modified IPS is absent on monolayer area, because the hole strongly interacts with substrate.

  9. Two-photon induced collagen cross-linking in bioartificial cardiac tissue

    Science.gov (United States)

    Kuetemeyer, Kai; Kensah, George; Heidrich, Marko; Meyer, Heiko; Martin, Ulrich; Gruh, Ina; Heisterkamp, Alexander

    2011-08-01

    Cardiac tissue engineering is a promising strategy for regenerative therapies to overcome the shortage of donor organs for transplantation. Besides contractile function, the stiffness of tissue engineered constructs is crucial to generate transplantable tissue surrogates with sufficient mechanical stability to withstand the high pressure present in the heart. Although several collagen cross-linking techniques have proven to be efficient in stabilizing biomaterials, they cannot be applied to cardiac tissue engineering, as cell death occurs in the treated area. Here, we present a novel method using femtosecond (fs) laser pulses to increase the stiffness of collagen-based tissue constructs without impairing cell viability. Raster scanning of the fs laser beam over riboflavin-treated tissue induced collagen cross-linking by two-photon photosensitized singlet oxygen production. One day post-irradiation, stress-strain measurements revealed increased tissue stiffness by around 40% being dependent on the fibroblast content in the tissue. At the same time, cells remained viable and fully functional as demonstrated by fluorescence imaging of cardiomyocyte mitochondrial activity and preservation of active contraction force. Our results indicate that two-photon induced collagen cross-linking has great potential for studying and improving artificially engineered tissue for regenerative therapies.

  10. Determining the quark charges by one and two photon processes

    International Nuclear Information System (INIS)

    Janah, A.

    1982-01-01

    Testable predictions are presented, which may be used to decide between the gauge theories of integer and fractionally charged quarks (icq and fcq). Two distinctive features of icq are exploited, namely (a) presence of color non-singlet components in weak and electromagnetic currents and (b) possible liberation of color non-singlet states above a threshold energy. Consequences are sought in lepton-hadron interaction processes, taking into account the known color-suppression effect. Single photon/weak-boson processes such as nuN → nuX distinguish between icq and fcq only above color-threshold. Experimental consequences of color-liberation in the above process are obtained. It is found that the gluon-parton contribution survives color-suppression to produce a significant rise in the structure functions when color-threshold is exceeded. Two-photon processes such as e + e - → e + e - + 2 jets distinguish between the two theories even below color threshold. To obtain the icq predictions for this process, one must take into account (a) the (momentum-dependent) color suppression and (b) the added contribution from pair production of charged gluons

  11. Higgs decay into two photons in a warped extra dimension

    Science.gov (United States)

    Hahn, Juliane; Hörner, Clara; Malm, Raoul; Neubert, Matthias; Novotny, Kristiane; Schmell, Christoph

    2014-05-01

    A detailed five-dimensional calculation of the Higgs-boson decay into two photons is performed in both the minimal and the custodially protected Randall-Sundrum (RS) model, where the Standard Model (SM) fields propagate in the bulk and the scalar sector lives on or near the IR brane. It is explicitly shown that the gauge invariance of the sum of diagrams involving bosonic fields in the SM also applies to the case of these RS scenarios. An exact expression for the amplitude in terms of the five-dimensional (5D) gauge-boson and fermion propagators is presented, which includes the full dependence on the Higgs-boson mass. Closed expressions for the 5D -boson propagators in the minimal and the custodial RS model are derived, which are valid to all orders in . In contrast to the fermion case, the result for the bosonic contributions to the amplitude is insensitive to the details of the localization of the Higgs profile on or near the IR brane. The various RS predictions for the rate of the process are compared with the latest LHC data, and exclusion regions for the RS model parameters are derived.

  12. Review of two-photon exchange in electron scattering

    Energy Technology Data Exchange (ETDEWEB)

    J. Arrington, P. G. Blunden, W. Melnitchouk

    2011-10-01

    We review the role of two-photon exchange (TPE) in electron-hadron scattering, focusing in particular on hadronic frameworks suitable for describing the low and moderate Q^2 region relevant to most experimental studies. We discuss the effects of TPE on the extraction of nucleon form factors and their role in the resolution of the proton electric to magnetic form factor ratio puzzle. The implications of TPE on various other observables, including neutron form factors, electroproduction of resonances and pions, and nuclear form factors, are summarized. Measurements seeking to directly identify TPE effects, such as through the angular dependence of polarization measurements, nonlinear epsilon contributions to the cross sections, and via e+p to e-p cross section ratios, are also outlined. In the weak sector, we describe the role of TPE and gamma-Z interference in parity-violating electron scattering, and assess their impact on the extraction of the strange form factors of the nucleon and the weak charge of the proton.

  13. Method for universal detection of two-photon polarization entanglement

    Science.gov (United States)

    Bartkiewicz, Karol; Horodecki, Paweł; Lemr, Karel; Miranowicz, Adam; Życzkowski, Karol

    2015-03-01

    Detecting and quantifying quantum entanglement of a given unknown state poses problems that are fundamentally important for quantum information processing. Surprisingly, no direct (i.e., without quantum tomography) universal experimental implementation of a necessary and sufficient test of entanglement has been designed even for a general two-qubit state. Here we propose an experimental method for detecting a collective universal witness, which is a necessary and sufficient test of two-photon polarization entanglement. It allows us to detect entanglement for any two-qubit mixed state and to establish tight upper and lower bounds on its amount. A different element of this method is the sequential character of its main components, which allows us to obtain relatively complicated information about quantum correlations with the help of simple linear-optical elements. As such, this proposal realizes a universal two-qubit entanglement test within the present state of the art of quantum optics. We show the optimality of our setup with respect to the minimal number of measured quantities.

  14. The determination of high-resolution spatio-temporal glacier motion fields from time-lapse sequences

    Science.gov (United States)

    Schwalbe, Ellen; Maas, Hans-Gerd

    2017-12-01

    This paper presents a comprehensive method for the determination of glacier surface motion vector fields at high spatial and temporal resolution. These vector fields can be derived from monocular terrestrial camera image sequences and are a valuable data source for glaciological analysis of the motion behaviour of glaciers. The measurement concepts for the acquisition of image sequences are presented, and an automated monoscopic image sequence processing chain is developed. Motion vector fields can be derived with high precision by applying automatic subpixel-accuracy image matching techniques on grey value patterns in the image sequences. Well-established matching techniques have been adapted to the special characteristics of the glacier data in order to achieve high reliability in automatic image sequence processing, including the handling of moving shadows as well as motion effects induced by small instabilities in the camera set-up. Suitable geo-referencing techniques were developed to transform image measurements into a reference coordinate system.The result of monoscopic image sequence analysis is a dense raster of glacier surface point trajectories for each image sequence. Each translation vector component in these trajectories can be determined with an accuracy of a few centimetres for points at a distance of several kilometres from the camera. Extensive practical validation experiments have shown that motion vector and trajectory fields derived from monocular image sequences can be used for the determination of high-resolution velocity fields of glaciers, including the analysis of tidal effects on glacier movement, the investigation of a glacier's motion behaviour during calving events, the determination of the position and migration of the grounding line and the detection of subglacial channels during glacier lake outburst floods.

  15. Effect of the coherent cancellation of the two-photon resonance on the generation of vacuum ultraviolet light by two-photon reasonantly enhanced four-wave mixing

    International Nuclear Information System (INIS)

    Payne, M.G.; Garrett, W.R.; Judish, J.P.; Wunderlich, R.

    1988-11-01

    Many of the most impressive demonstrations of the efficient generation of vacuum ultraviolet (VUV) light have made use of two- photon resonantly enhanced four-wave mixing to generate light at ω/sub VUV/ = 2ω/sub L1/ +- ω/sub L2/. The two-photon resonance state is coupled to the ground state both by two photons from the first laser, or by a photon from the second laser and one from the generated VUV beam. We show here that these two coherent pathways destructively interfere once the second laser is made sufficiently intense, thereby leading to an important limiting effect on the achievable conversion efficiency. 4 refs

  16. Recent results on two-photon physics from Tasso and a review of measurements of the two-photon total cross section

    International Nuclear Information System (INIS)

    Kolanoski, H.

    1983-03-01

    Recent results on two-photon physics from the Tasso experiment are presented: the measurement of the two-photon production of Kanti K with the determination of the #betta##betta#-width of the f'(1515), an analysis of the angular correlations in the reaction #betta##betta#->rho 0 rho 0 ->π + π - π + π - and the observation of a narrow structure in the four pion mass spectrum around 2.1 GeV. In a separate part the experimental results on the total cross section for hadron production by two photons are reviewed. (orig.)

  17. Femtosecond laser subsurface scleral treatment in cadaver human sclera and evaluation using two-photon and confocal microscopy

    Science.gov (United States)

    Sun, Hui; Fan, Zhongwei; Yan, Ying; Lian, Fuqiang; Kurtz, Ron; Juhasz, Tibor

    2016-03-01

    Glaucoma is the second-leading cause of blindness worldwide and is often associated with elevated intraocular pressure (IOP). Partial-thickness drainage channels can be created with femtosecond laser in the translucent sclera for the potential treatment of glaucoma. We demonstrate the creation of partial-thickness subsurface drainage channels with the femtosecond laser in the cadaver human eyeballs and describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. A femtosecond laser operating at a wavelength of 1700 nm was scanned along a rectangular raster pattern to create the partial thickness subsurface drainage channels in the sclera of cadaver human eyes. Analysis of the dimensions and location of these channels is important in understanding their effects. We describe the application of two-photon microscopy and confocal microscopy for noninvasive imaging of the femtosecond laser created partial-thickness scleral channels in cadaver human eyes. High-resolution images, hundreds of microns deep in the sclera, were obtained to allow determination of the shape and dimension of such partial thickness subsurface scleral channels. Our studies suggest that the confocal and two-photon microscopy can be used to investigate femtosecond-laser created partial-thickness drainage channels in the sclera of cadaver human eyes.

  18. Distribution of quantum information between an atom and two photons

    International Nuclear Information System (INIS)

    Weber, Bernhard

    2008-01-01

    The construction of networks consisting of optically interconnected processing units is a promising way to scale up quantum information processing systems. To store quantum information, single trapped atoms are among the most proven candidates. By placing them in high finesse optical resonators, a bidirectional information exchange between the atoms and photons becomes possible with, in principle, unit efficiency. Such an interface between stationary and ying qubits constitutes a possible node of a future quantum network. The results presented in this thesis demonstrate the prospects of a quantum interface consisting of a single atom trapped within the mode of a high-finesse optical cavity. In a two-step process, we distribute entanglement between the stored atom and two subsequently emitted single photons. The long atom trapping times achieved in the system together with the high photon collection efficiency of the cavity make the applied protocol in principle deterministic, allowing for the creation of an entangled state at the push of a button. Running the protocol on this quasi-stationary quantum interface, the internal state of the atom is entangled with the polarization state of a single emitted photon. The entanglement is generated by driving a vacuum-stimulated Raman adiabatic passage between states of the coupled atom-cavity system. In a second process, the atomic part of the entangled state is mapped onto a second emitted photon using a similar technique and resulting in a polarization-entangled two-photon state. To verify and characterize the photon-photon entanglement, we measured a violation of a Bell inequality and performed a full quantum state tomography. The results prove the prior atom-photon entanglement and demonstrate a quantum information transfer between the atom and the two emitted photons. This reflects the advantages of a high-finesse cavity as a quantum interface in future quantum networks. (orig.)

  19. Dynamical modeling of pulsed two-photon interference

    International Nuclear Information System (INIS)

    Fischer, Kevin A; Lagoudakis, Konstantinos G; Vučković, Jelena; Müller, Kai

    2016-01-01

    Single-photon sources are at the heart of quantum-optical networks, with their uniquely quantum emission and phenomenon of two-photon interference allowing for the generation and transfer of nonclassical states. Although a few analytical methods have been briefly investigated for describing pulsed single-photon sources, these methods apply only to either perfectly ideal or at least extremely idealized sources. Here, we present the first complete picture of pulsed single-photon sources by elaborating how to numerically and fully characterize non-ideal single-photon sources operating in a pulsed regime. In order to achieve this result, we make the connection between quantum Monte-Carlo simulations, experimental characterizations, and an extended form of the quantum regression theorem. We elaborate on how an ideal pulsed single-photon source is connected to its photocount distribution and its measured degree of second- and first-order optical coherence. By doing so, we provide a description of the relationship between instantaneous source correlations and the typical experimental interferometers (Hanbury-Brown and Twiss, Hong–Ou–Mandel, and Mach–Zehnder) used to characterize such sources. Then, we use these techniques to explore several prototypical quantum systems and their non-ideal behaviors. As an example numerical result, we show that for the most popular single-photon source—a resonantly excited two-level system—its error probability is directly related to its excitation pulse length. We believe that the intuition gained from these representative systems and characters can be used to interpret future results with more complicated source Hamiltonians and behaviors. Finally, we have thoroughly documented our simulation methods with contributions to the Quantum Optics Toolbox in Python in order to make our work easily accessible to other scientists and engineers. (paper)

  20. Distribution of quantum information between an atom and two photons

    Energy Technology Data Exchange (ETDEWEB)

    Weber, Bernhard

    2008-11-03

    The construction of networks consisting of optically interconnected processing units is a promising way to scale up quantum information processing systems. To store quantum information, single trapped atoms are among the most proven candidates. By placing them in high finesse optical resonators, a bidirectional information exchange between the atoms and photons becomes possible with, in principle, unit efficiency. Such an interface between stationary and ying qubits constitutes a possible node of a future quantum network. The results presented in this thesis demonstrate the prospects of a quantum interface consisting of a single atom trapped within the mode of a high-finesse optical cavity. In a two-step process, we distribute entanglement between the stored atom and two subsequently emitted single photons. The long atom trapping times achieved in the system together with the high photon collection efficiency of the cavity make the applied protocol in principle deterministic, allowing for the creation of an entangled state at the push of a button. Running the protocol on this quasi-stationary quantum interface, the internal state of the atom is entangled with the polarization state of a single emitted photon. The entanglement is generated by driving a vacuum-stimulated Raman adiabatic passage between states of the coupled atom-cavity system. In a second process, the atomic part of the entangled state is mapped onto a second emitted photon using a similar technique and resulting in a polarization-entangled two-photon state. To verify and characterize the photon-photon entanglement, we measured a violation of a Bell inequality and performed a full quantum state tomography. The results prove the prior atom-photon entanglement and demonstrate a quantum information transfer between the atom and the two emitted photons. This reflects the advantages of a high-finesse cavity as a quantum interface in future quantum networks. (orig.)