Sample records for quantitative time-lapse microscopy

  1. Comparative evaluation of performance measures for shading correction in time-lapse fluorescence microscopy.

    PubMed

    Liu, L; Kan, A; Leckie, C; Hodgkin, P D

    2017-04-01

    Time-lapse fluorescence microscopy is a valuable technology in cell biology, but it suffers from the inherent problem of intensity inhomogeneity due to uneven illumination or camera nonlinearity, known as shading artefacts. This will lead to inaccurate estimates of single-cell features such as average and total intensity. Numerous shading correction methods have been proposed to remove this effect. In order to compare the performance of different methods, many quantitative performance measures have been developed. However, there is little discussion about which performance measure should be generally applied for evaluation on real data, where the ground truth is absent. In this paper, the state-of-the-art shading correction methods and performance evaluation methods are reviewed. We implement 10 popular shading correction methods on two artificial datasets and four real ones. In order to make an objective comparison between those methods, we employ a number of quantitative performance measures. Extensive validation demonstrates that the coefficient of joint variation (CJV) is the most applicable measure in time-lapse fluorescence images. Based on this measure, we have proposed a novel shading correction method that performs better compared to well-established methods for a range of real data tested. © 2016 The Authors Journal of Microscopy © 2016 Royal Microscopical Society.

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

    PubMed

    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.

  3. Time-lapse microscopy of lung endothelial cells under hypoxia

    NASA Astrophysics Data System (ADS)

    Mehrvar, Shima; Ghanian, Zahra; Kondouri, Ganesh; Camara, Amadou S.; Ranji, Mahsa

    2017-02-01

    Objective: This study utilizes fluorescence microscopy to assess the effect of the oxygen tension on the production of reactive oxygen species (ROS) in mitochondria of fetal pulmonary artery endothelial cells (FPAECs). Introduction: Hypoxia is a severe oxygen stress, which mostly causes irreversible injury in lung cells. However, in some studies, it is reported that hypoxia decreases the severity of injuries. In this study, ROS production level was examined in hypoxic FPAECs treated with pentachlorophenol (PCP, uncoupler). This work was accomplished by monitoring and quantifying the changes in the level of the produced ROS in hypoxic cells before and after PCP treatment. Materials and methods: The dynamic of the mitochondrial ROS production in two groups of FPAECs was measured over time using time-lapse microscopy. For the first group, cells were incubated in 3% hypoxic condition for 2 hours and then continuously were exposed to hypoxic condition for imaging as well. For the second group, cells were incubated in normal oxygen condition. Time lapse images of the cells loaded with Mito-SOX (ROS indicator) were acquired, and the red fluorescence intensity profile of the cells was calculated. Changes in the level of the fluorescence intensity profile while they are treated with PCP indicates the dynamics of the ROS level. Results: The intensity profiles of the PCP-treated cells in the first group showed 47% lower ROS production rate than the PCP-treated cells in the second group. Conclusion: Time lapse microscopy revealed that hypoxic cells have lower ROS generation while treated with PCP. Therefore, this result suggests that hypoxia decreased electron transport chain activity in uncoupled chain.

  4. [Embryo selection in IVF/ICSI cycles using time-lapse microscopy and the clinical outcomes].

    PubMed

    Chen, Minghao; Huang, Jun; Zhong, Ying; Quan, Song

    2015-12-01

    To compare the clinical outcomes of embryos selected using time-lapse microscopy and traditional morphological method in IVF/ICSI cycles and evaluate the clinical value of time-lapse microscopy in early embryo monitoring and selection. e retrospectively analyzed the clinical data of 139 IVF/ICSI cycles with embryo selection based on time-lapse monitoring (TLM group, n=68) and traditional morphological method (control group, n=71). The βHCG-positive rate, clinical pregnancy rate and embryo implantation rate were compared between the 2 groups. Subgroup analysis was performed in view of female patients age and the fertilization type. The βHCG-positive rate, clinical pregnancy rate and implantation rate were 66.2%, 61.8% and 47.1% in TLM group, significantly higher than those in the control group (47.9%, 43.7% and 30.3%, respectively; P<0.05). Compared with patients below 30 years of age, patients aged between 31 and 35 years benefited more from time-lapse monitoring with improved clinical outcomes. time-lapse monitoring significantly increased the βHCG-positive rate, clinical pregnancy rate and implantation rate for patients undergoing IVF cycles, but not for those undergoing ICSI or TESA cycles. Compared with those selected using traditional morphological method, the embryos selected with time-lapse microscopy have better clinical outcomes, especially in older patients (31-35 years of age) and in IVF cycles.

  5. Measuring fast gene dynamics in single cells with time-lapse luminescence microscopy

    PubMed Central

    Mazo-Vargas, Anyimilehidi; Park, Heungwon; Aydin, Mert; Buchler, Nicolas E.

    2014-01-01

    Time-lapse fluorescence microscopy is an important tool for measuring in vivo gene dynamics in single cells. However, fluorescent proteins are limited by slow chromophore maturation times and the cellular autofluorescence or phototoxicity that arises from light excitation. An alternative is luciferase, an enzyme that emits photons and is active upon folding. The photon flux per luciferase is significantly lower than that for fluorescent proteins. Thus time-lapse luminescence microscopy has been successfully used to track gene dynamics only in larger organisms and for slower processes, for which more total photons can be collected in one exposure. Here we tested green, yellow, and red beetle luciferases and optimized substrate conditions for in vivo luminescence. By combining time-lapse luminescence microscopy with a microfluidic device, we tracked the dynamics of cell cycle genes in single yeast with subminute exposure times over many generations. Our method was faster and in cells with much smaller volumes than previous work. Fluorescence of an optimized reporter (Venus) lagged luminescence by 15–20 min, which is consistent with its known rate of chromophore maturation in yeast. Our work demonstrates that luciferases are better than fluorescent proteins at faithfully tracking the underlying gene expression. PMID:25232010

  6. Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy.

    PubMed

    Di Sante, Gabriele; Casimiro, Mathew C; Pestell, Timothy G; Pestell, Richard G

    2016-05-04

    Time-lapse video microscopy can be defined as the real time imaging of living cells. This technique relies on the collection of images at different time points. Time intervals can be set through a computer interface that controls the microscope-integrated camera. This kind of microscopy requires both the ability to acquire very rapid events and the signal generated by the observed cellular structure during these events. After the images have been collected, a movie of the entire experiment is assembled to show the dynamic of the molecular events of interest. Time-lapse video microscopy has a broad range of applications in the biomedical research field and is a powerful and unique tool for following the dynamics of the cellular events in real time. Through this technique, we can assess cellular events such as migration, division, signal transduction, growth, and death. Moreover, using fluorescent molecular probes we are able to mark specific molecules, such as DNA, RNA or proteins and follow them through their molecular pathways and functions. Time-lapse video microscopy has multiple advantages, the major one being the ability to collect data at the single-cell level, that make it a unique technology for investigation in the field of cell biology. However, time-lapse video microscopy has limitations that can interfere with the acquisition of high quality images. Images can be compromised by both external factors; temperature fluctuations, vibrations, humidity and internal factors; pH, cell motility. Herein, we describe a protocol for the dynamic acquisition of a specific protein, Parkin, fused with the enhanced yellow fluorescent protein (EYFP) in order to track the selective removal of damaged mitochondria, using a time-lapse video microscopy approach.

  7. Measuring single-cell gene expression dynamics in bacteria using fluorescence time-lapse microscopy

    PubMed Central

    Young, Jonathan W; Locke, James C W; Altinok, Alphan; Rosenfeld, Nitzan; Bacarian, Tigran; Swain, Peter S; Mjolsness, Eric; Elowitz, Michael B

    2014-01-01

    Quantitative single-cell time-lapse microscopy is a powerful method for analyzing gene circuit dynamics and heterogeneous cell behavior. We describe the application of this method to imaging bacteria by using an automated microscopy system. This protocol has been used to analyze sporulation and competence differentiation in Bacillus subtilis, and to quantify gene regulation and its fluctuations in individual Escherichia coli cells. The protocol involves seeding and growing bacteria on small agarose pads and imaging the resulting microcolonies. Images are then reviewed and analyzed using our laboratory's custom MATLAB analysis code, which segments and tracks cells in a frame-to-frame method. This process yields quantitative expression data on cell lineages, which can illustrate dynamic expression profiles and facilitate mathematical models of gene circuits. With fast-growing bacteria, such as E. coli or B. subtilis, image acquisition can be completed in 1 d, with an additional 1–2 d for progressing through the analysis procedure. PMID:22179594

  8. Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy

    PubMed Central

    Di Sante, Gabriele; Casimiro, Mathew C.; Pestell, Timothy G.; Pestell, Richard G.

    2016-01-01

    Time-lapse video microscopy can be defined as the real time imaging of living cells. This technique relies on the collection of images at different time points. Time intervals can be set through a computer interface that controls the microscope-integrated camera. This kind of microscopy requires both the ability to acquire very rapid events and the signal generated by the observed cellular structure during these events. After the images have been collected, a movie of the entire experiment is assembled to show the dynamic of the molecular events of interest. Time-lapse video microscopy has a broad range of applications in the biomedical research field and is a powerful and unique tool for following the dynamics of the cellular events in real time. Through this technique, we can assess cellular events such as migration, division, signal transduction, growth, and death. Moreover, using fluorescent molecular probes we are able to mark specific molecules, such as DNA, RNA or proteins and follow them through their molecular pathways and functions. Time-lapse video microscopy has multiple advantages, the major one being the ability to collect data at the single-cell level, that make it a unique technology for investigation in the field of cell biology. However, time-lapse video microscopy has limitations that can interfere with the acquisition of high quality images. Images can be compromised by both external factors; temperature fluctuations, vibrations, humidity and internal factors; pH, cell motility. Herein, we describe a protocol for the dynamic acquisition of a specific protein, Parkin, fused with the enhanced yellow fluorescent protein (EYFP) in order to track the selective removal of damaged mitochondria, using a time-lapse video microscopy approach. PMID:27168174

  9. 3D lens-free time-lapse microscopy for 3D cell culture

    NASA Astrophysics Data System (ADS)

    Berdeu, Anthony; Momey, Fabien; Laperrousaz, Bastien; Bordy, Thomas; Gidrol, Xavier; Dinten, Jean-Marc; Picollet-D'hahan, Nathalie; Allier, Cédric

    2017-07-01

    We propose a new imaging platform based on lens-free time-lapse microscopy for 3D cell culture and its dedicated algorithm lying on a fully 3D regularized inverse problem approach. First 3D+t results are presented

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

    PubMed

    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. Time-lapse microscopy patent upheld in Europe.

    PubMed

    Pearce, David

    2017-02-01

    A case for revoking Stanford University's European patent 2430454 on time-lapse microscopy was set out in Reproductive BioMedicine Online by Sterckx et al. in 2014, on the grounds that the patent claimed a method of diagnosis that was excluded under a provision of the European Patent Convention. An opposition at the European Patent Office in which this ground was raised has recently concluded with a decision that the patent is not excluded from patentability under European patent law and is to be upheld. An appeal from this decision has been filed, but the possibility of the decision being overturned is, in this author's opinion, very limited. Copyright © 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  12. Quantitative Analysis of Complex Glioma Cell Migration on Electrospun Polycaprolactone Using Time-Lapse Microscopy

    PubMed Central

    Johnson, Jed; Nowicki, M. Oskar; Lee, Carol H.; Chiocca, E. Antonio; Viapiano, Mariano S.; Lawler, Sean E.

    2009-01-01

    Malignant gliomas are the most common tumors originating within the central nervous system and account for over 15,000 deaths annually in the United States. The median survival for glioblastoma, the most common and aggressive of these tumors, is only 14 months. Therapeutic strategies targeting glioma cells migrating away from the tumor core are currently hampered by the difficulty of reproducing migration in the neural parenchyma in vitro. We utilized a tissue engineering approach to develop a physiologically relevant model of glioma cell migration. This revealed that glioma cells display dramatic differences in migration when challenged by random versus aligned electrospun poly-ɛ-caprolactone nanofibers. Cells on aligned fibers migrated at an effective velocity of 4.2 ± 0.39 μm/h compared to 0.8 ± 0.08 μm/h on random fibers, closely matching in vivo models and prior observations of glioma spread in white versus gray matter. Cells on random fibers exhibited extension along multiple fiber axes that prevented net motion; aligned fibers promoted a fusiform morphology better suited to infiltration. Time-lapse microscopy revealed that the motion of individual cells was complex and was influenced by cell cycle and local topography. Glioma stem cell–containing neurospheres seeded on random fibers did not show cell detachment and retained their original shape; on aligned fibers, cells detached and migrated in the fiber direction over a distance sixfold greater than the perpendicular direction. This chemically and physically flexible model allows time-lapse analysis of glioma cell migration while recapitulating in vivo cell morphology, potentially allowing identification of physiological mediators and pharmacological inhibitors of invasion. PMID:19199562

  13. Time-lapse microscopy and image analysis in basic and clinical embryo development research.

    PubMed

    Wong, C; Chen, A A; Behr, B; Shen, S

    2013-02-01

    Mammalian preimplantation embryo development is a complex process in which the exact timing and sequence of events are as essential as the accurate execution of the events themselves. Time-lapse microscopy (TLM) is an ideal tool to study this process since the ability to capture images over time provides a combination of morphological, dynamic and quantitative information about developmental events. Here, we systematically review the application of TLM in basic and clinical embryo research. We identified all relevant preimplantation embryo TLM studies published in English up to May 2012 using PubMed and Google Scholar. We then analysed the technical challenges involved in embryo TLM studies and how these challenges may be overcome with technological innovations. Finally, we reviewed the different types of TLM embryo studies, with a special focus on how TLM can benefit clinical assisted reproduction. Although new parameters predictive of embryo development potential may be discovered and used clinically to potentially increase the success rate of IVF, adopting TLM to routine clinical practice will require innovations in both optics and image analysis. Combined with such innovations, TLM may provide embryologists and clinicians with an important tool for making critical decisions in assisted reproduction. In this review, we perform a literature search of all published early embryo development studies that used time-lapse microscopy (TLM). From the literature, we discuss the benefits of TLM over traditional time-point analysis, as well as the technical difficulties and solutions involved in implementing TLM for embryo studies. We further discuss research that has successfully derived non-invasive markers that may increase the success rate of assisted reproductive technologies, primarily IVF. Most notably, we extend our discussion to highlight important considerations for the practical use of TLM in research and clinical settings. Copyright © 2012 Reproductive

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

    PubMed

    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

  15. A new protocol for functional analysis of adipogenesis using reverse transfection technology and time-lapse video microscopy.

    PubMed

    Grönniger, Elke; Wessel, Sonja; Kühn, Sonja Christin; Söhle, Jörn; Wenck, Horst; Stäb, Franz; Winnefeld, Marc

    2010-07-01

    Since the worldwide increase in obesity represents a growing challenge for healthcare systems, research focusing on fat cell metabolism has become a focal point of interest. Here, we describe a small interfering RNA (siRNA)-technology-based screening method to study fat cell differentiation in human primary preadipocytes that could be further developed towards an automated middle-throughput screening procedure. First, we established optimal conditions for the reverse transfection of human primary preadipocytes demonstrating that an efficient reverse transfection of preadipocytes is technically feasible. Aligning the processes of reverse transfection and fat cell differentiation utilizing peroxisome proliferator-activated receptor gamma (PPAR gamma)-siRNA, we showed that preadipocyte differentiation was suppressed by knock-down of PPAR gamma, the key regulator of fat cell differentiation. The use of fluorescently labelled fatty acids in combination with fluorescence time-lapse microscopy over a longer period of time enabled us to quantify the PPAR gamma phenotype. Additionally, our data demonstrate that reverse transfection of human cultured preadipocytes with TIP60 (HIV-1 Tat-interacting protein 60)-siRNA lead to a TIP60 knock-down and subsequently inhibits fat cell differentiation, suggesting a role of this protein in human adipogenesis. In conclusion, we established a protocol that allows for an efficient functional and time-dependent analysis by quantitative time-lapse microscopy to identify novel adipogenesis-associated genes.

  16. Image-based characterization of thrombus formation in time-lapse DIC microscopy

    PubMed Central

    Brieu, Nicolas; Navab, Nassir; Serbanovic-Canic, Jovana; Ouwehand, Willem H.; Stemple, Derek L.; Cvejic, Ana; Groher, Martin

    2012-01-01

    The characterization of thrombus formation in time-lapse DIC microscopy is of increased interest for identifying genes which account for atherothrombosis and coronary artery diseases (CADs). In particular, we are interested in large-scale studies on zebrafish, which result in large amount of data, and require automatic processing. In this work, we present an image-based solution for the automatized extraction of parameters quantifying the temporal development of thrombotic plugs. Our system is based on the joint segmentation of thrombotic and aortic regions over time. This task is made difficult by the low contrast and the high dynamic conditions observed in vivo DIC microscopic scenes. Our key idea is to perform this segmentation by distinguishing the different motion patterns in image time series rather than by solving standard image segmentation tasks in each image frame. Thus, we are able to compensate for the poor imaging conditions. We model motion patterns by energies based on the idea of dynamic textures, and regularize the model by two prior energies on the shape of the aortic region and on the topological relationship between the thrombus and the aorta. We demonstrate the performance of our segmentation algorithm by qualitative and quantitative experiments on synthetic examples as well as on real in vivo microscopic sequences. PMID:22482997

  17. Quantitative analysis of random migration of cells using time-lapse video microscopy.

    PubMed

    Jain, Prachi; Worthylake, Rebecca A; Alahari, Suresh K

    2012-05-13

    Cell migration is a dynamic process, which is important for embryonic development, tissue repair, immune system function, and tumor invasion (1, 2). During directional migration, cells move rapidly in response to an extracellular chemotactic signal, or in response to intrinsic cues (3) provided by the basic motility machinery. Random migration occurs when a cell possesses low intrinsic directionality, allowing the cells to explore their local environment. Cell migration is a complex process, in the initial response cell undergoes polarization and extends protrusions in the direction of migration (2). Traditional methods to measure migration such as the Boyden chamber migration assay is an easy method to measure chemotaxis in vitro, which allows measuring migration as an end point result. However, this approach neither allows measurement of individual migration parameters, nor does it allow to visualization of morphological changes that cell undergoes during migration. Here, we present a method that allows us to monitor migrating cells in real time using video - time lapse microscopy. Since cell migration and invasion are hallmarks of cancer, this method will be applicable in studying cancer cell migration and invasion in vitro. Random migration of platelets has been considered as one of the parameters of platelet function (4), hence this method could also be helpful in studying platelet functions. This assay has the advantage of being rapid, reliable, reproducible, and does not require optimization of cell numbers. In order to maintain physiologically suitable conditions for cells, the microscope is equipped with CO(2) supply and temperature thermostat. Cell movement is monitored by taking pictures using a camera fitted to the microscope at regular intervals. Cell migration can be calculated by measuring average speed and average displacement, which is calculated by Slidebook software.

  18. Time-lapse microscopy using smartphone with augmented reality markers.

    PubMed

    Baek, Dongyoub; Cho, Sungmin; Yun, Kyungwon; Youn, Keehong; Bang, Hyunwoo

    2014-04-01

    A prototype system that replaces the conventional time-lapse imaging in microscopic inspection for use with smartphones is presented. Existing time-lapse imaging requires a video data feed between a microscope and a computer that varies depending on the type of image grabber. Even with proper hardware setups, a series of tedious and repetitive tasks is still required to relocate to the region-of-interest (ROI) of the specimens. In order to simplify the system and improve the efficiency of time-lapse imaging tasks, a smartphone-based platform utilizing microscopic augmented reality (μ-AR) markers is proposed. To evaluate the feasibility and efficiency of the proposed system, a user test was designed and performed, measuring the elapse time for a trial of the task starting from the execution of the application software to the completion of restoring and imaging of an ROI saved in advance. The results of the user test showed that the average elapse time was 65.3 ± 15.2 s with 6.86 ± 3.61 μm of position error and 0.08 ± 0.40 degrees of angle error. This indicates that the time-lapse imaging task was accomplished rapidly with a high level of accuracy. Thus, simplification of both the system and the task was achieved via our proposed system. Copyright © 2014 Wiley Periodicals, Inc.

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

    PubMed

    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.

  20. Sequential processing of quantitative phase images for the study of cell behaviour in real-time digital holographic microscopy.

    PubMed

    Zikmund, T; Kvasnica, L; Týč, M; Křížová, A; Colláková, J; Chmelík, R

    2014-11-01

    Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The study of the cell is based on extraction of the dynamic data on cell behaviour from the time-lapse sequence of the phase images. However, the phase images are affected by the phase aberrations that make the analysis particularly difficult. This is because the phase deformation is prone to change during long-term experiments. Here, we present a novel algorithm for sequential processing of living cells phase images in a time-lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least-squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. All these procedures are performed automatically and applied immediately after obtaining every single phase image. This property of the algorithm is important for real-time cell quantitative phase imaging and instantaneous control of the course of the experiment by playback of the recorded sequence up to actual time. Such operator's intervention is a forerunner of process automation derived from image analysis. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off-axis holographic microscope. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

  1. Time-lapse microscopy patent upheld in Europe: response to Pearce.

    PubMed

    Sterckx, Sigrid; Cockbain, Julian; Pennings, Guido

    2017-02-01

    In this piece, we comment on the article by Pearce earlier in this journal. As Pearce correctly points out, what is fundamentally at issue in ESHRE et al's opposition to Stanford University's European patent on time-lapse microscopy is whether an exclusion from patentability, here of methods of medical diagnosis, should be interpreted narrowly or not. In the present case, the dominant piece of case law from the European Patent Office (EPO) gives a narrow interpretation of what a method of diagnosis must be in order not to be patentable. In their submissions to the EPO, ESHRE et al. have argued that this narrow interpretation is unfounded and incorrect. Copyright © 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  2. Quantitative Estimation of Seismic Velocity Changes Using Time-Lapse Seismic Data and Elastic-Wave Sensitivity Approach

    NASA Astrophysics Data System (ADS)

    Denli, H.; Huang, L.

    2008-12-01

    Quantitative monitoring of reservoir property changes is essential for safe geologic carbon sequestration. Time-lapse seismic surveys have the potential to effectively monitor fluid migration in the reservoir that causes geophysical property changes such as density, and P- and S-wave velocities. We introduce a novel method for quantitative estimation of seismic velocity changes using time-lapse seismic data. The method employs elastic sensitivity wavefields, which are the derivatives of elastic wavefield with respect to density, P- and S-wave velocities of a target region. We derive the elastic sensitivity equations from analytical differentiations of the elastic-wave equations with respect to seismic-wave velocities. The sensitivity equations are coupled with the wave equations in a way that elastic waves arriving in a target reservoir behave as a secondary source to sensitivity fields. We use a staggered-grid finite-difference scheme with perfectly-matched layers absorbing boundary conditions to simultaneously solve the elastic-wave equations and the elastic sensitivity equations. By elastic-wave sensitivities, a linear relationship between relative seismic velocity changes in the reservoir and time-lapse seismic data at receiver locations can be derived, which leads to an over-determined system of equations. We solve this system of equations using a least- square method for each receiver to obtain P- and S-wave velocity changes. We validate the method using both surface and VSP synthetic time-lapse seismic data for a multi-layered model and the elastic Marmousi model. Then we apply it to the time-lapse field VSP data acquired at the Aneth oil field in Utah. A total of 10.5K tons of CO2 was injected into the oil reservoir between the two VSP surveys for enhanced oil recovery. The synthetic and field data studies show that our new method can quantitatively estimate changes in seismic velocities within a reservoir due to CO2 injection/migration.

  3. Optofluidic time-stretch quantitative phase microscopy.

    PubMed

    Guo, Baoshan; Lei, Cheng; Wu, Yi; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Lee, Sangwook; Isozaki, Akihiro; Li, Ming; Jiang, Yiyue; Yasumoto, Atsushi; Di Carlo, Dino; Tanaka, Yo; Yatomi, Yutaka; Ozeki, Yasuyuki; Goda, Keisuke

    2018-03-01

    Innovations in optical microscopy have opened new windows onto scientific research, industrial quality control, and medical practice over the last few decades. One of such innovations is optofluidic time-stretch quantitative phase microscopy - an emerging method for high-throughput quantitative phase imaging that builds on the interference between temporally stretched signal and reference pulses by using dispersive properties of light in both spatial and temporal domains in an interferometric configuration on a microfluidic platform. It achieves the continuous acquisition of both intensity and phase images with a high throughput of more than 10,000 particles or cells per second by overcoming speed limitations that exist in conventional quantitative phase imaging methods. Applications enabled by such capabilities are versatile and include characterization of cancer cells and microalgal cultures. In this paper, we review the principles and applications of optofluidic time-stretch quantitative phase microscopy and discuss its future perspective. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. A time-series method for automated measurement of changes in mitotic and interphase duration from time-lapse movies.

    PubMed

    Sigoillot, Frederic D; Huckins, Jeremy F; Li, Fuhai; Zhou, Xiaobo; Wong, Stephen T C; King, Randall W

    2011-01-01

    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.

  5. Label-free quantitative cell division monitoring of endothelial cells by digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Kemper, Björn; Bauwens, Andreas; Vollmer, Angelika; Ketelhut, Steffi; Langehanenberg, Patrik; Müthing, Johannes; Karch, Helge; von Bally, Gert

    2010-05-01

    Digital holographic microscopy (DHM) enables quantitative multifocus phase contrast imaging for nondestructive technical inspection and live cell analysis. Time-lapse investigations on human brain microvascular endothelial cells demonstrate the use of DHM for label-free dynamic quantitative monitoring of cell division of mother cells into daughter cells. Cytokinetic DHM analysis provides future applications in toxicology and cancer research.

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

    ERIC Educational Resources Information Center

    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…

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

    NASA Astrophysics Data System (ADS)

    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

  8. Gliding Motility of Babesia bovis Merozoites Visualized by Time-Lapse Video Microscopy

    PubMed Central

    Asada, Masahito; Goto, Yasuyuki; Yahata, Kazuhide; Yokoyama, Naoaki; Kawai, Satoru; Inoue, Noboru; Kaneko, Osamu; Kawazu, Shin-ichiro

    2012-01-01

    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. PMID:22506073

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

    PubMed

    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/.

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

    PubMed

    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.

  11. Time-lapse cinemicrography and scanning electron microscopy of platelet formation by megakaryocytes.

    PubMed

    Haller, C J; Radley, J M

    1983-01-01

    The surface architecture of megakaryocytes undergoing platelet formation in vitro has been examined by time-lapse cinemicrography and scanning electron microscopy. Fragments of mouse bone marrow were placed in culture medium and incubated at 37 degrees C. After several hours mature megakaryocytes migrated out of the marrow and some underwent shape changes so that they eventually appeared as a relatively small central body, housing the nucleus, from which emerged a number of thin processes which resembled platelet chains. Scanning electron microscopy showed that initially the megakaryocyte surface was ruffled but with development of processes it became smoother. Circumferential folds of small amplitude were found on the surface of developing constrictions which separated putative platelets. It is thought they may be associated with the mechanism of extension, but could have a role in establishing the topography of membrane components. Rupture of the chains and release of platelets was not observed; this permits the number of putative platelets formed by individual megakaryocytes to be determined. The putative platelets exhibited features common to circulating platelets when exposed to a glass surface including the development of pseudopodia and, eventually, flattening on to the surface.

  12. Significantly improved precision of cell migration analysis in time-lapse video microscopy through use of a fully automated tracking system

    PubMed Central

    2010-01-01

    Background Cell motility is a critical parameter in many physiological as well as pathophysiological processes. In time-lapse video microscopy, manual cell tracking remains the most common method of analyzing migratory behavior of cell populations. In addition to being labor-intensive, this method is susceptible to user-dependent errors regarding the selection of "representative" subsets of cells and manual determination of precise cell positions. Results We have quantitatively analyzed these error sources, demonstrating that manual cell tracking of pancreatic cancer cells lead to mis-calculation of migration rates of up to 410%. In order to provide for objective measurements of cell migration rates, we have employed multi-target tracking technologies commonly used in radar applications to develop fully automated cell identification and tracking system suitable for high throughput screening of video sequences of unstained living cells. Conclusion We demonstrate that our automatic multi target tracking system identifies cell objects, follows individual cells and computes migration rates with high precision, clearly outperforming manual procedures. PMID:20377897

  13. Time-lapse microscopy and image processing for stem cell research: modeling cell migration

    NASA Astrophysics Data System (ADS)

    Gustavsson, Tomas; Althoff, Karin; Degerman, Johan; Olsson, Torsten; Thoreson, Ann-Catrin; Thorlin, Thorleif; Eriksson, Peter

    2003-05-01

    This paper presents hardware and software procedures for automated cell tracking and migration modeling. A time-lapse microscopy system equipped with a computer controllable motorized stage was developed. The performance of this stage was improved by incorporating software algorithms for stage motion displacement compensation and auto focus. The microscope is suitable for in-vitro stem cell studies and allows for multiple cell culture image sequence acquisition. This enables comparative studies concerning rate of cell splits, average cell motion velocity, cell motion as a function of cell sample density and many more. Several cell segmentation procedures are described as well as a cell tracking algorithm. Statistical methods for describing cell migration patterns are presented. In particular, the Hidden Markov Model (HMM) was investigated. Results indicate that if the cell motion can be described as a non-stationary stochastic process, then the HMM can adequately model aspects of its dynamic behavior.

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

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

    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.

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

    PubMed

    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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27886235

  17. Large-scale time-lapse microscopy of Oct4 expression in human embryonic stem cell colonies.

    PubMed

    Bhadriraju, Kiran; Halter, Michael; Amelot, Julien; Bajcsy, Peter; Chalfoun, Joe; Vandecreme, Antoine; Mallon, Barbara S; Park, Kye-Yoon; Sista, Subhash; Elliott, John T; Plant, Anne L

    2016-07-01

    Identification and quantification of the characteristics of stem cell preparations is critical for understanding stem cell biology and for the development and manufacturing of stem cell based therapies. We have developed image analysis and visualization software that allows effective use of time-lapse microscopy to provide spatial and dynamic information from large numbers of human embryonic stem cell colonies. To achieve statistically relevant sampling, we examined >680 colonies from 3 different preparations of cells over 5days each, generating a total experimental dataset of 0.9 terabyte (TB). The 0.5 Giga-pixel images at each time point were represented by multi-resolution pyramids and visualized using the Deep Zoom Javascript library extended to support viewing Giga-pixel images over time and extracting data on individual colonies. We present a methodology that enables quantification of variations in nominally-identical preparations and between colonies, correlation of colony characteristics with Oct4 expression, and identification of rare events. Copyright © 2016. Published by Elsevier B.V.

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

    PubMed

    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.

  19. Early Events in Insulin Fibrillization Studied by Time-Lapse Atomic Force Microscopy

    PubMed Central

    Podestà, Alessandro; Tiana, Guido; Milani, Paolo; Manno, Mauro

    2006-01-01

    The importance of understanding the mechanism of protein aggregation into insoluble amyloid fibrils lies not only in its medical consequences, but also in its more basic properties of self-organization. The discovery that a large number of uncorrelated proteins can form, under proper conditions, structurally similar fibrils has suggested that the underlying mechanism is a general feature of polypeptide chains. In this work, we address the early events preceding amyloid fibril formation in solutions of zinc-free human insulin incubated at low pH and high temperature. Here, we show by time-lapse atomic force microscopy that a steady-state distribution of protein oligomers with a quasiexponential tail is reached within a few minutes after heating. This metastable phase lasts for a few hours, until fibrillar aggregates are observable. Although for such complex systems different aggregation mechanisms can occur simultaneously, our results indicate that the prefibrillar phase is mainly controlled by a simple coagulation-evaporation kinetic mechanism, in which concentration acts as a critical parameter. These experimental facts, along with the kinetic model used, suggest a critical role for thermal concentration fluctuations in the process of fibril nucleation. PMID:16239333

  20. Automated segmentation and tracking of non-rigid objects in time-lapse microscopy videos of polymorphonuclear neutrophils.

    PubMed

    Brandes, Susanne; Mokhtari, Zeinab; Essig, Fabian; Hünniger, Kerstin; Kurzai, Oliver; Figge, Marc Thilo

    2015-02-01

    Time-lapse microscopy is an important technique to study the dynamics of various biological processes. The labor-intensive manual analysis of microscopy videos is increasingly replaced by automated segmentation and tracking methods. These methods are often limited to certain cell morphologies and/or cell stainings. In this paper, we present an automated segmentation and tracking framework that does not have these restrictions. In particular, our framework handles highly variable cell shapes and does not rely on any cell stainings. Our segmentation approach is based on a combination of spatial and temporal image variations to detect moving cells in microscopy videos. This method yields a sensitivity of 99% and a precision of 95% in object detection. The tracking of cells consists of different steps, starting from single-cell tracking based on a nearest-neighbor-approach, detection of cell-cell interactions and splitting of cell clusters, and finally combining tracklets using methods from graph theory. The segmentation and tracking framework was applied to synthetic as well as experimental datasets with varying cell densities implying different numbers of cell-cell interactions. We established a validation framework to measure the performance of our tracking technique. The cell tracking accuracy was found to be >99% for all datasets indicating a high accuracy for connecting the detected cells between different time points. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

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

    PubMed Central

    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

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

    NASA Astrophysics Data System (ADS)

    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.

  4. Microfluidic device enabled quantitative time-lapse microscopic-photography for phenotyping vegetative and reproductive phases in Fusarium virguliforme, which is pathogenic to soybean.

    PubMed

    Marshall, Jill; Qiao, Xuan; Baumbach, Jordan; Xie, Jingyu; Dong, Liang; Bhattacharyya, Madan K

    2017-03-15

    Time-lapse microscopic-photography allows in-depth phenotyping of microorganisms. Here we report development of such a system using a microfluidic device, generated from polydimethylsiloxane and glass slide, placed on a motorized stage of a microscope for conducting time-lapse microphotography of multiple observations in 20 channels simultaneously. We have demonstrated the utility of the device in studying growth, germination and sporulation in Fusarium virguliforme that causes sudden death syndrome in soybean. To measure the growth differences, we developed a polyamine oxidase fvpo1 mutant in this fungus that fails to grow in minimal medium containing polyamines as the sole nitrogen source. Using this system, we demonstrated that the conidiospores of the pathogen take an average of five hours to germinate. During sporulation, it takes an average of 10.5 h for a conidiospore to mature and get detached from its conidiophore for the first time. Conidiospores are developed in a single conidiophore one after another. The microfluidic device enabled quantitative time-lapse microphotography reported here should be suitable for screening compounds, peptides, micro-organisms to identify fungitoxic or antimicrobial agents for controlling serious plant pathogens. The device could also be applied in identifying suitable target genes for host-induced gene silencing in pathogens for generating novel disease resistance in crop plants.

  5. Quantitative microscopy uncovers ploidy changes during mitosis in live Drosophila embryos and their effect on nuclear size.

    PubMed

    Puah, Wee Choo; Chinta, Rambabu; Wasser, Martin

    2017-03-15

    Time-lapse microscopy is a powerful tool to investigate cellular and developmental dynamics. In Drosophila melanogaster , it can be used to study division cycles in embryogenesis. To obtain quantitative information from 3D time-lapse data and track proliferating nuclei from the syncytial stage until gastrulation, we developed an image analysis pipeline consisting of nuclear segmentation, tracking, annotation and quantification. Image analysis of maternal-haploid ( mh ) embryos revealed that a fraction of haploid syncytial nuclei fused to give rise to nuclei of higher ploidy (2n, 3n, 4n). Moreover, nuclear densities in mh embryos at the mid-blastula transition varied over threefold. By tracking synchronized nuclei of different karyotypes side-by-side, we show that DNA content determines nuclear growth rate and size in early interphase, while the nuclear to cytoplasmic ratio constrains nuclear growth during late interphase. mh encodes the Drosophila ortholog of human Spartan, a protein involved in DNA damage tolerance. To explore the link between mh and chromosome instability, we fluorescently tagged Mh protein to study its subcellular localization. We show Mh-mKO2 localizes to nuclear speckles that increase in numbers as nuclei expand in interphase. In summary, quantitative microscopy can provide new insights into well-studied genes and biological processes. © 2017. Published by The Company of Biologists Ltd.

  6. Time-lapse imaging of disease progression in deep brain areas using fluorescence microendoscopy

    PubMed Central

    Barretto, Robert P. J.; Ko, Tony H.; Jung, Juergen C.; Wang, Tammy J.; Capps, George; Waters, Allison C.; Ziv, Yaniv; Attardo, Alessio; Recht, Lawrence; Schnitzer, Mark J.

    2013-01-01

    The combination of intravital microscopy and animal models of disease has propelled studies of disease mechanisms and treatments. However, many disorders afflict tissues inaccessible to light microscopy in live subjects. Here we introduce cellular-level time-lapse imaging deep within the live mammalian brain by one- and two-photon fluorescence microendoscopy over multiple weeks. Bilateral imaging sites allowed longitudinal comparisons within individual subjects, including of normal and diseased tissues. Using this approach we tracked CA1 hippocampal pyramidal neuron dendrites in adult mice, revealing these dendrites' extreme stability (>8,000 day mean lifetime) and rare examples of their structural alterations. To illustrate disease studies, we tracked deep lying gliomas by observing tumor growth, visualizing three-dimensional vasculature structure, and determining microcirculatory speeds. Average erythrocyte speeds in gliomas declined markedly as the disease advanced, notwithstanding significant increases in capillary diameters. Time-lapse microendoscopy will be applicable to studies of numerous disorders, including neurovascular, neurological, cancerous, and trauma-induced conditions. PMID:21240263

  7. In situ monitoring of corrosion mechanisms and phosphate inhibitor surface deposition during corrosion of zinc-magnesium-aluminium (ZMA) alloys using novel time-lapse microscopy.

    PubMed

    Sullivan, James; Cooze, Nathan; Gallagher, Callum; Lewis, Tom; Prosek, Tomas; Thierry, Dominique

    2015-01-01

    In situ time-lapse optical microscopy was used to examine the microstructural corrosion mechanisms in three zinc-magnesium-aluminium (ZMA) alloy coated steels immersed in 1% NaCl pH 7. Preferential corrosion of MgZn(2) lamellae within the eutectic phases was observed in all the ZMA alloys followed by subsequent dissolution of Zn rich phases. The total extent and rate of corrosion, measured using time-lapse image analysis and scanning vibrating electrode technique (SVET) estimated mass loss, decreased as Mg and Al alloying additions were increased up to a level of 3 wt% Mg and 3.7 wt% Al. This was probably due to the increased presence of MgO and Al(2)O(3) at the alloy surface retarding the kinetics of cathodic oxygen reduction. The addition of 1 × 10(-2) mol dm(-3) Na(3)PO(4) to 1% NaCl pH 7 had a dramatic influence on the corrosion mechanism for a ZMA with passivation of anodic sites through phosphate precipitation observed using time-lapse image analysis. Intriguing rapid precipitation of filamentous phosphate was also observed and it is postulated that these filaments nucleate and grow due to super saturation effects. Polarisation experiments showed that the addition of 1 × 10(-2) mol dm(-3) Na(3)PO(4) to the 1% NaCl electrolyte promoted an anodic shift of 50 mV in open circuit potential for the ZMA alloy with a reduction in anodic current of 2.5 orders of magnitude suggesting that it was acting primarily as an anodic inhibitor supporting the inferences from the time-lapse investigations. These phosphate additions resulted in a 98% reduction in estimated mass loss as measured by SVET demonstrating the effectiveness of phosphate inhibitors for this alloy system.

  8. Time-lapse imaging of mitosis after siRNA transfection.

    PubMed

    Mackay, Douglas R; Ullman, Katharine S; Rodesch, Christopher K

    2010-06-06

    Changes in cellular organization and chromosome dynamics that occur during mitosis are tightly coordinated to ensure accurate inheritance of genomic and cellular content. Hallmark events of mitosis, such as chromosome movement, can be readily tracked on an individual cell basis using time-lapse fluorescence microscopy of mammalian cell lines expressing specific GFP-tagged proteins. In combination with RNAi-based depletion, this can be a powerful method for pinpointing the stage(s) of mitosis where defects occur after levels of a particular protein have been lowered. In this protocol, we present a basic method for assessing the effect of depleting a potential mitotic regulatory protein on the timing of mitosis. Cells are transfected with siRNA, placed in a stage-top incubation chamber, and imaged using an automated fluorescence microscope. We describe how to use software to set up a time-lapse experiment, how to process the image sequences to make either still-image montages or movies, and how to quantify and analyze the timing of mitotic stages using a cell-line expressing mCherry-tagged histone H2B. Finally, we discuss important considerations for designing a time-lapse experiment. This strategy is complementary to other approaches and offers the advantages of 1) sensitivity to changes in kinetics that might not be observed when looking at cells as a population and 2) analysis of mitosis without the need to synchronize the cell cycle using drug treatments. The visual information from such imaging experiments not only allows the sub-stages of mitosis to be assessed, but can also provide unexpected insight that would not be apparent from cell cycle analysis by FACS.

  9. Time-lapse analysis of gravitropism in Ceratodon protonemata

    NASA Technical Reports Server (NTRS)

    Young, J. C.; Sack, F. D.

    1992-01-01

    The tip cell of the protonema of the moss Ceratodon purpureus (Hedw.) Brid. is negatively gravitropic when grown in the dark on supplemented agar. Gravitropism, plastid distribution, and plastid movement were studied in living cells using time-lapse video microscopy and infrared light. A wrong-way (downward) curvature preceded upward curvature and was detected as early as 2 minutes after reorientation. Upward curvature began 30-45 minutes after reorientation to the horizontal. Cell division temporarily reversed upward curvature, but did not inhibit wrong-way curvature. Since significant amyloplast sedimentation always occurred before the start of upward curvature, it is possible that these amyloplasts function as statoliths for upward curvature. However, no significant amyloplast sedimentation occurred before wrong-way curvature. Thus, this early phase of gravitropism cannot require plastid sedimentation for gravity sensing. Most plastids moved within and between zones, and plastid zonation was highly dynamic. Plastids moved toward the apex and toward the base of the cell at rates much slower than cytoplasmic streaming. Despite the dynamic nature of plastid movement and zonation, during upward curvature the distance between sedimented plastids and the apex stayed constant. Time-lapse analysis has revealed intriguing events not readily seen previously using destructive sampling.

  10. QueSST Time-lapse

    NASA Image and Video Library

    2017-02-27

    Quiet Supersonic Technology (QueSST) X-plane in the 8x6 Supersonic Wind Tunnel at NASA Glenn Research Center. This time-lapse shows the model support structure buildup and balance checkout as well as the installation of the model in the test section.

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

    USGS Publications Warehouse

    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

  12. Visualization of living terminal hypertrophic chondrocytes of growth plate cartilage in situ by differential interference contrast microscopy and time-lapse cinematography.

    PubMed

    Farnum, C E; Turgai, J; Wilsman, N J

    1990-09-01

    The functional unit within the growth plate consists of a column of chondrocytes that passes through a sequence of phases including proliferation, hypertrophy, and death. It is important to our understanding of the biology of the growth plate to determine if distal hypertrophic cells are viable, highly differentiated cells with the potential of actively controlling terminal events of endochondral ossification prior to their death at the chondro-osseous junction. This study for the first time reports on the visualization of living hypertrophic chondrocytes in situ, including the terminal hypertrophic chondrocyte. Chondrocytes in growth plate explants are visualized using rectified differential interference contrast microscopy. We record and measure, using time-lapse cinematography, the rate of movement of subcellular organelles at the limit of resolution of this light microscopy system. Control experiments to assess viability of hypertrophic chondrocytes include coincubating organ cultures with the intravital dye fluorescein diacetate to assess the integrity of the plasma membrane and cytoplasmic esterases. In this system, all hypertrophic chondrocytes, including the very terminal chondrocyte, exist as rounded, fully hydrated cells. By the criteria of intravital dye staining and organelle movement, distal hypertrophic chondrocytes are identical to chondrocytes in the proliferative and early hypertrophic cell zones.

  13. 3D Time-lapse Imaging and Quantification of Mitochondrial Dynamics

    NASA Astrophysics Data System (ADS)

    Sison, Miguel; Chakrabortty, Sabyasachi; Extermann, Jérôme; Nahas, Amir; James Marchand, Paul; Lopez, Antonio; Weil, Tanja; Lasser, Theo

    2017-02-01

    We present a 3D time-lapse imaging method for monitoring mitochondrial dynamics in living HeLa cells based on photothermal optical coherence microscopy and using novel surface functionalization of gold nanoparticles. The biocompatible protein-based biopolymer coating contains multiple functional groups which impart better cellular uptake and mitochondria targeting efficiency. The high stability of the gold nanoparticles allows continuous imaging over an extended time up to 3000 seconds without significant cell damage. By combining temporal autocorrelation analysis with a classical diffusion model, we quantify mitochondrial dynamics and cast these results into 3D maps showing the heterogeneity of diffusion parameters across the whole cell volume.

  14. Time-Lapse Measurement of Wellbore Integrity

    NASA Astrophysics Data System (ADS)

    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

  15. Single-cell time-lapse imaging of intracellular O2 in response to metabolic inhibition and mitochondrial cytochrome-c release.

    PubMed

    Düssmann, Heiko; Perez-Alvarez, Sergio; Anilkumar, Ujval; Papkovsky, Dmitri B; Prehn, Jochen Hm

    2017-06-01

    The detection of intracellular molecular oxygen (O 2 ) levels is important for understanding cell physiology, cell death, and drug effects, and has recently been improved with the development of oxygen-sensitive probes that are compatible with live cell time-lapse microscopy. We here provide a protocol for the use of the nanoparticle probe MitoImage-MM2 to monitor intracellular oxygen levels by confocal microscopy under baseline conditions, in response to mitochondrial toxins, and following mitochondrial cytochrome-c release. We demonstrate that the MitoImage-MM2 probe, which embeds Pt(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorophenyl)-porphyrin as oxygen sensor and poly(9,9-dioctylfluorene) as an O 2 -independent component, enables quantitative, ratiometric time-lapse imaging of intracellular O 2 . Multiplexing with tetra-methyl-rhodamine-methyl ester in HeLa cervical cancer cells showed significant increases in intracellular O 2 accompanied by strong mitochondrial depolarization when respiratory chain complexes III or IV were inhibited by Antimycin A or sodium azide, respectively, and when cells were maintained at 'physiological' tissue O 2 levels (5% O 2 ). Multiplexing also allowed us to monitor intracellular O 2 during the apoptotic signaling process of mitochondrial outer membrane permeabilization in HeLa expressing cytochrome-c-eGFP, and demonstrated that mitochondria post cytochrome-c release are able to retain their capacity to respire at physiological O 2 despite a decrease in mitochondrial membrane potential.

  16. Single-cell time-lapse imaging of intracellular O2 in response to metabolic inhibition and mitochondrial cytochrome-c release

    PubMed Central

    Düssmann, Heiko; Perez-Alvarez, Sergio; Anilkumar, Ujval; Papkovsky, Dmitri B; Prehn, Jochen HM

    2017-01-01

    The detection of intracellular molecular oxygen (O2) levels is important for understanding cell physiology, cell death, and drug effects, and has recently been improved with the development of oxygen-sensitive probes that are compatible with live cell time-lapse microscopy. We here provide a protocol for the use of the nanoparticle probe MitoImage-MM2 to monitor intracellular oxygen levels by confocal microscopy under baseline conditions, in response to mitochondrial toxins, and following mitochondrial cytochrome-c release. We demonstrate that the MitoImage-MM2 probe, which embeds Pt(II)-5,10,15,20-tetrakis-(2,3,4,5,6–pentafluorophenyl)-porphyrin as oxygen sensor and poly(9,9-dioctylfluorene) as an O2-independent component, enables quantitative, ratiometric time-lapse imaging of intracellular O2. Multiplexing with tetra-methyl-rhodamine-methyl ester in HeLa cervical cancer cells showed significant increases in intracellular O2 accompanied by strong mitochondrial depolarization when respiratory chain complexes III or IV were inhibited by Antimycin A or sodium azide, respectively, and when cells were maintained at ‘physiological’ tissue O2 levels (5% O2). Multiplexing also allowed us to monitor intracellular O2 during the apoptotic signaling process of mitochondrial outer membrane permeabilization in HeLa expressing cytochrome-c-eGFP, and demonstrated that mitochondria post cytochrome-c release are able to retain their capacity to respire at physiological O2 despite a decrease in mitochondrial membrane potential. PMID:28569778

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

    NASA Astrophysics Data System (ADS)

    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.

  18. In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

    PubMed Central

    Oudin, Madeleine Julie; Doherty, Patrick; Lalli, Giovanna

    2013-01-01

    The subventricular zone (SVZ) is one of the main neurogenic niches in the postnatal brain. Here, neural progenitors proliferate and give rise to neuroblasts able to move along the rostral migratory stream (RMS) towards the olfactory bulb (OB). This long-distance migration is required for the subsequent maturation of newborn neurons in the OB, but the molecular mechanisms regulating this process are still unclear. Investigating the signaling pathways controlling neuroblast motility may not only help understand a fundamental step in neurogenesis, but also have therapeutic regenerative potential, given the ability of these neuroblasts to target brain sites affected by injury, stroke, or degeneration. In this manuscript we describe a detailed protocol for in vivo postnatal electroporation and subsequent time-lapse imaging of neuroblast migration in the mouse RMS. Postnatal electroporation can efficiently transfect SVZ progenitor cells, which in turn generate neuroblasts migrating along the RMS. Using confocal spinning disk time-lapse microscopy on acute brain slice cultures, neuroblast migration can be monitored in an environment closely resembling the in vivo condition. Moreover, neuroblast motility can be tracked and quantitatively analyzed. As an example, we describe how to use in vivo postnatal electroporation of a GFP-expressing plasmid to label and visualize neuroblasts migrating along the RMS. Electroporation of shRNA or CRE recombinase-expressing plasmids in conditional knockout mice employing the LoxP system can also be used to target genes of interest. Pharmacological manipulation of acute brain slice cultures can be performed to investigate the role of different signaling molecules in neuroblast migration. By coupling in vivo electroporation with time-lapse imaging, we hope to understand the molecular mechanisms controlling neuroblast motility and contribute to the development of novel approaches to promote brain repair. PMID:24326479

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

    NASA Astrophysics Data System (ADS)

    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

  20. Evaluation of automated time-lapse microscopy for assessment of in vitro activity of antibiotics.

    PubMed

    Ungphakorn, Wanchana; Malmberg, Christer; Lagerbäck, Pernilla; Cars, Otto; Nielsen, Elisabet I; Tängdén, Thomas

    2017-01-01

    This study aimed to evaluate the potential of a new time-lapse microscopy based method (oCelloScope) to efficiently assess the in vitro antibacterial effects of antibiotics. Two E. coli and one P. aeruginosa strain were exposed to ciprofloxacin, colistin, ertapenem and meropenem in 24-h experiments. Background corrected absorption (BCA) derived from the oCelloScope was used to detect bacterial growth. The data obtained with the oCelloScope were compared with those of the automated Bioscreen C method and standard time-kill experiments and a good agreement in results was observed during 6-24h of experiments. Viable counts obtained at 1, 4, 6 and 24h during oCelloScope and Bioscreen C experiments were well correlated with the corresponding BCA and optical density (OD) data. Initial antibacterial effects during the first 6h of experiments were difficult to detect with the automated methods due to their high detection limits (approximately 10 5 CFU/mL for oCelloScope and 10 7 CFU/mL for Bioscreen C), the inability to distinguish between live and dead bacteria and early morphological changes of bacteria during exposure to ciprofloxacin, ertapenem and meropenem. Regrowth was more frequently detected in time-kill experiments, possibly related to the larger working volume with an increased risk of pre-existing or emerging resistance. In comparison with Bioscreen C, the oCelloScope provided additional information on bacterial growth dynamics in the range of 10 5 to 10 7 CFU/mL and morphological features. In conclusion, the oCelloScope would be suitable for detection of in vitro effects of antibiotics, especially when a large number of regimens need to be tested. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Johnson, Timothy C.; Versteeg, Roelof; Day-Lewis, Frederick D.

    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 ERTmore » 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 surfacebased ERT in conjunction with limited field sampling to improve

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

    PubMed Central

    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

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

    PubMed

    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.

  4. Influence of salinomycin treatment on division and movement of individual cancer cells cultured in normoxia or hypoxia evaluated with time-lapse digital holographic microscopy

    PubMed Central

    Kamlund, Sofia; Strand, Daniel; Janicke, Birgit; Alm, Kersti; Oredsson, Stina

    2017-01-01

    ABSTRACT Most studies on new cancer drugs are based on population-derived data, where the absence of response of a small population may pass unnoticed. Thus, individual longitudinal tracking of cells is important for the future development of efficient cancer treatments. We have used digital holographic microscopy to track individual JIMT-1 human breast cancer cells and L929 mouse fibroblast cultivated in normoxia or hypoxia. In addition, JIMT-1 cells were treated with salinomycin, a cancer stem cell targeting compound. Three-day time-lapse movies were captured and individual cells were analysed with respect to cell division (cell cycle length) and cell movement. Comparing population-doubling time derived from population-based growth curves and individual cell cycle time data from time-lapse movies show that the former hide a sub-population of dividing cells. Salinomycin treatment increased the motility of cells, however, this motility did not result in an increased distant migration i.e. the cells increased their local movement. MCF-7 breast cancer cells showed similar motility behaviour as salinomycin-treated JIMT-1 cells. We suggest that combining features, such as motility and migration, can be used to distinguish cancer cells with mesenchymal (JIMT-1) and epithelial (MCF-7) features. The data clearly emphasize the importance of longitudinal cell tracking to understand the biology of individual cells under different conditions. PMID:28933990

  5. Real time quantitative phase microscopy based on single-shot transport of intensity equation (ssTIE) method

    NASA Astrophysics Data System (ADS)

    Yu, Wei; Tian, Xiaolin; He, Xiaoliang; Song, Xiaojun; Xue, Liang; Liu, Cheng; Wang, Shouyu

    2016-08-01

    Microscopy based on transport of intensity equation provides quantitative phase distributions which opens another perspective for cellular observations. However, it requires multi-focal image capturing while mechanical and electrical scanning limits its real time capacity in sample detections. Here, in order to break through this restriction, real time quantitative phase microscopy based on single-shot transport of the intensity equation method is proposed. A programmed phase mask is designed to realize simultaneous multi-focal image recording without any scanning; thus, phase distributions can be quantitatively retrieved in real time. It is believed the proposed method can be potentially applied in various biological and medical applications, especially for live cell imaging.

  6. A-3 Construction Time Lapse

    NASA Technical Reports Server (NTRS)

    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.

  7. Nanoparticles for the delivery of zoledronic acid to prostate cancer cells: A comparative analysis through time lapse video-microscopy technique

    PubMed Central

    Schiraldi, Chiara; Zappavigna, Silvia; D' Agostino, Antonella; Porto, Stefania; Gaito, Ornella; Lusa, Sara; Lamberti, Monica; De Rosa, Mario; De Rosa, Giuseppe; Caraglia, Michele

    2014-01-01

    Time-lapse live cell imaging is a powerful tool for studying the responses of cells to drugs. Zoledronic acid (ZOL) is the most potent aminobiphosphonate able to induce cell growth inhibition at very low concentrations. The lack of clear evidence of ZOL-induced anti-cancer effects is likely due to its unfavorable pharmacokinetic profile. The use of nanotechnology-based formulations allows overcoming these limitations in ZOL pharmaco-distribution. Recently, stealth liposomes (LIPOs) and new self-assembly PEGylated nanoparticles (NPs) encapsulating ZOL were developed. Both the delivery systems showed promising anticancer activity in vitro and in vivo. In this work, we investigated the cytostatic effect of these novel formulations (LIPOs and NPs) compared with free ZOL on 2 different prostate cancer cell lines, PC 3 and DU 145 and on prostate epithelial primary cells EPN using time lapse video-microscopy (TLVM). In PC3 cells, free ZOL showed a significant anti-proliferative effect but this effect was lower than that induced by LIPOs and NPs encapsulating ZOL; moreover, LIPO-ZOL was more potent in inducing growth inhibition than NP-ZOL. On the other hand, LIPO-ZOL slightly enhanced the free ZOL activity on growth inhibition of DU 145, while the anti-proliferative effect of NP-ZOL was not statistically relevant. These novel formulations did not induce anti-proliferative effects on EPN cells. Finally, we evaluated cytotoxic effects on DU145 where, LIPO-ZOL induced the highest cytotoxicity compared with NP-ZOL and free ZOL. In conclusion, ZOL can be transformed in a powerful anticancer agent, if administered with nanotechnology-based formulations without damaging the healthy tissues. PMID:25482949

  8. Joint inversion of time-lapse VSP data for monitoring CO2 injection at the Farnsworth EOR field in Texas

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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

  10. Fluorescence Time-lapse Imaging of the Complete S. venezuelae Life Cycle Using a Microfluidic Device.

    PubMed

    Schlimpert, Susan; Flärdh, Klas; Buttner, Mark

    2016-02-28

    Live-cell imaging of biological processes at the single cell level has been instrumental to our current understanding of the subcellular organization of bacterial cells. However, the application of time-lapse microscopy to study the cell biological processes underpinning development in the sporulating filamentous bacteria Streptomyces has been hampered by technical difficulties. Here we present a protocol to overcome these limitations by growing the new model species, Streptomyces venezuelae, in a commercially available microfluidic device which is connected to an inverted fluorescence widefield microscope. Unlike the classical model species, Streptomyces coelicolor, S. venezuelae sporulates in liquid, allowing the application of microfluidic growth chambers to cultivate and microscopically monitor the cellular development and differentiation of S. venezuelae over long time periods. In addition to monitoring morphological changes, the spatio-temporal distribution of fluorescently labeled target proteins can also be visualized by time-lapse microscopy. Moreover, the microfluidic platform offers the experimental flexibility to exchange the culture medium, which is used in the detailed protocol to stimulate sporulation of S. venezuelae in the microfluidic chamber. Images of the entire S. venezuelae life cycle are acquired at specific intervals and processed in the open-source software Fiji to produce movies of the recorded time-series.

  11. Fluorescence Time-lapse Imaging of the Complete S. venezuelae Life Cycle Using a Microfluidic Device

    PubMed Central

    Schlimpert, Susan; Flärdh, Klas; Buttner, Mark

    2016-01-01

    Live-cell imaging of biological processes at the single cell level has been instrumental to our current understanding of the subcellular organization of bacterial cells. However, the application of time-lapse microscopy to study the cell biological processes underpinning development in the sporulating filamentous bacteria Streptomyces has been hampered by technical difficulties. Here we present a protocol to overcome these limitations by growing the new model species, Streptomyces venezuelae, in a commercially available microfluidic device which is connected to an inverted fluorescence widefield microscope. Unlike the classical model species, Streptomyces coelicolor, S. venezuelae sporulates in liquid, allowing the application of microfluidic growth chambers to cultivate and microscopically monitor the cellular development and differentiation of S. venezuelae over long time periods. In addition to monitoring morphological changes, the spatio-temporal distribution of fluorescently labeled target proteins can also be visualized by time-lapse microscopy. Moreover, the microfluidic platform offers the experimental flexibility to exchange the culture medium, which is used in the detailed protocol to stimulate sporulation of S. venezuelae in the microfluidic chamber. Images of the entire S. venezuelae life cycle are acquired at specific intervals and processed in the open-source software Fiji to produce movies of the recorded time-series. PMID:26967231

  12. Quantitative dispersion microscopy

    PubMed Central

    Fu, Dan; Choi, Wonshik; Sung, Yongjin; Yaqoob, Zahid; Dasari, Ramachandra R.; Feld, Michael

    2010-01-01

    Refractive index dispersion is an intrinsic optical property and a useful source of contrast in biological imaging studies. In this report, we present the first dispersion phase imaging of living eukaryotic cells. We have developed quantitative dispersion microscopy based on the principle of quantitative phase microscopy. The dual-wavelength quantitative phase microscope makes phase measurements at 310 nm and 400 nm wavelengths to quantify dispersion (refractive index increment ratio) of live cells. The measured dispersion of living HeLa cells is found to be around 1.088, which agrees well with that measured directly for protein solutions using total internal reflection. This technique, together with the dry mass and morphology measurements provided by quantitative phase microscopy, could prove to be a useful tool for distinguishing different types of biomaterials and studying spatial inhomogeneities of biological samples. PMID:21113234

  13. Time-Lapse, in Situ Imaging of Ice Crystal Growth Using Confocal Microscopy

    PubMed Central

    2016-01-01

    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. PMID:27917410

  14. Time-Lapse, in Situ Imaging of Ice Crystal Growth Using Confocal Microscopy.

    PubMed

    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.

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

    NASA Astrophysics Data System (ADS)

    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.

  16. Rapid antimicrobial susceptibility testing of clinical isolates by digital time-lapse microscopy.

    PubMed

    Fredborg, M; Rosenvinge, F S; Spillum, E; Kroghsbo, S; Wang, M; Sondergaard, T E

    2015-12-01

    Rapid antimicrobial susceptibility testing (AST) is essential for early and appropriate therapy. Methods with short detection time enabling same-day treatment optimisation are highly favourable. In this study, we evaluated the potential of a digital time-lapse microscope system, the oCelloScope system, to perform rapid AST. The oCelloScope system demonstrated a very high accuracy (96% overall agreement) when determining the resistance profiles of four reference strains, nine clinical isolates, including multi-drug-resistant isolates, and three positive blood cultures. AST of clinical isolates (168 antimicrobial agent-organism combinations) demonstrated 3.6% minor, no major and 1.2% very major errors of the oCelloScope system compared to conventional susceptibility testing, as well as a rapid and correct phenotypic detection of strains with methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum β-lactamase (ESBL) profiles. The net average time-to-result was 108 min, with 95% of the results being available within 180 min. In conclusion, this study strongly indicates that the oCelloScope system holds considerable potential as an accurate and sensitive AST method with short time-to-result, enabling same-day targeted antimicrobial therapy, facilitating antibiotic stewardship and better patient management. A full-scale validation of the oCelloScope system including more isolates is necessary to assess the impact of using it for AST.

  17. Time-lapse Raman imaging of osteoblast differentiation

    PubMed Central

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

    2015-01-01

    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. PMID:26211729

  18. Time-lapse Raman imaging of osteoblast differentiation

    NASA Astrophysics Data System (ADS)

    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.

  19. Intelligent data analysis to model and understand live cell time-lapse sequences.

    PubMed

    Paterson, Allan; Ashtari, M; Ribé, D; Stenbeck, G; Tucker, A

    2012-01-01

    One important aspect of cellular function, which is at the basis of tissue homeostasis, is the delivery of proteins to their correct destinations. Significant advances in live cell microscopy have allowed tracking of these pathways by following the dynamics of fluorescently labelled proteins in living cells. This paper explores intelligent data analysis techniques to model the dynamic behavior of proteins in living cells as well as to classify different experimental conditions. We use a combination of decision tree classification and hidden Markov models. In particular, we introduce a novel approach to "align" hidden Markov models so that hidden states from different models can be cross-compared. Our models capture the dynamics of two experimental conditions accurately with a stable hidden state for control data and multiple (less stable) states for the experimental data recapitulating the behaviour of particle trajectories within live cell time-lapse data. In addition to having successfully developed an automated framework for the classification of protein transport dynamics from live cell time-lapse data our model allows us to understand the dynamics of a complex trafficking pathway in living cells in culture.

  20. A semi-Markov model for mitosis segmentation in time-lapse phase contrast microscopy image sequences of stem cell populations.

    PubMed

    Liu, An-An; Li, Kang; Kanade, Takeo

    2012-02-01

    We propose a semi-Markov model trained in a max-margin learning framework for mitosis event segmentation in large-scale time-lapse phase contrast microscopy image sequences of stem cell populations. Our method consists of three steps. First, we apply a constrained optimization based microscopy image segmentation method that exploits phase contrast optics to extract candidate subsequences in the input image sequence that contains mitosis events. Then, we apply a max-margin hidden conditional random field (MM-HCRF) classifier learned from human-annotated mitotic and nonmitotic sequences to classify each candidate subsequence as a mitosis or not. Finally, a max-margin semi-Markov model (MM-SMM) trained on manually-segmented mitotic sequences is utilized to reinforce the mitosis classification results, and to further segment each mitosis into four predefined temporal stages. The proposed method outperforms the event-detection CRF model recently reported by Huh as well as several other competing methods in very challenging image sequences of multipolar-shaped C3H10T1/2 mesenchymal stem cells. For mitosis detection, an overall precision of 95.8% and a recall of 88.1% were achieved. For mitosis segmentation, the mean and standard deviation for the localization errors of the start and end points of all mitosis stages were well below 1 and 2 frames, respectively. In particular, an overall temporal location error of 0.73 ± 1.29 frames was achieved for locating daughter cell birth events.

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

    NASA Astrophysics Data System (ADS)

    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

  2. Using Digital Time-Lapse Videos to Teach Geomorphic Processes to Undergraduates

    NASA Astrophysics Data System (ADS)

    Clark, D. H.; Linneman, S. R.; Fuller, J.

    2004-12-01

    We demonstrate the use of relatively low-cost, computer-based digital imagery to create time-lapse videos of two distinct geomorphic processes in order to help students grasp the significance of the rates, styles, and temporal dependence of geologic phenomena. Student interviews indicate that such videos help them to understand the relationship between processes and landform development. Time-lapse videos have been used extensively in some sciences (e.g., biology - http://sbcf.iu.edu/goodpract/hangarter.html, meteorology - http://www.apple.com/education/hed/aua0101s/meteor/, chemistry - http://www.chem.yorku.ca/profs/hempsted/chemed/home.html) to demonstrate gradual processes that are difficult for many students to visualize. Most geologic processes are slower still, and are consequently even more difficult for students to grasp, yet time-lapse videos are rarely used in earth science classrooms. The advent of inexpensive web-cams and computers provides a new means to explore the temporal dimension of earth surface processes. To test the use of time-lapse videos in geoscience education, we are developing time-lapse movies that record the evolution of two landforms: a stream-table delta and a large, natural, active landslide. The former involves well-known processes in a controlled, repeatable laboratory experiment, whereas the latter tracks the developing dynamics of an otherwise poorly understood slope failure. The stream-table delta is small and grows in ca. 2 days; we capture a frame on an overhead web-cam every 3 minutes. Before seeing the video, students are asked to hypothesize how the delta will grow through time. The final time-lapse video, ca. 20-80 MB, elegantly shows channel migration, progradation rates, and formation of major geomorphic elements (topset, foreset, bottomset beds). The web-cam can also be "zoomed-in" to show smaller-scale processes, such as bedload transfer, and foreset slumping. Post-lab tests and interviews with students indicate that

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

    PubMed

    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

  4. Time-lapse photogrammetry in geomorphic studies

    NASA Astrophysics Data System (ADS)

    Eltner, Anette; Kaiser, Andreas

    2017-04-01

    Image based approaches to reconstruct the earth surface (Structure from Motion - SfM) are establishing as a standard technology for high resolution topographic data. This is amongst other advantages due to the comparatively ease of use and flexibility of data generation. Furthermore, the increased spatial resolution led to its implementation at a vast range of applications from sub-mm to tens-of-km scale. Almost fully automatic calculation of referenced digital elevation models allows for a significant increase of temporal resolution, as well, potentially up to sub-second scales. Thereby, the setup of a time-lapse multi-camera system is necessary and different aspects need to be considered: The camera array has to be temporary stable or potential movements need to be compensated by temporary stable reference targets/areas. The stability of the internal camera geometry has to be considered due to a usually significantly lower amount of images of the scene, and thus redundancy for parameter estimation, compared to more common SfM applications. Depending on the speed of surface change, synchronisation has to be very accurate. Due to the usual application in the field, changing environmental conditions important for lighting and visual range are also crucial factors to keep in mind. Besides these important considerations much potential is comprised by time-lapse photogrammetry. The integration of multi-sensor systems, e.g. using thermal cameras, enables the potential detection of other processes not visible with RGB-images solely. Furthermore, the implementation of low-cost sensors allows for a significant increase of areal coverage and their setup at locations, where a loss of the system cannot be ruled out. The usage of micro-computers offers smart camera triggering, e.g. acquiring images with increased frequency controlled by a rainfall-triggered sensor. In addition these micro-computers can enable on-site data processing, e.g. recognition of increased surface

  5. Dynamic quantitative analysis of adherent cell cultures by means of lens-free video microscopy

    NASA Astrophysics Data System (ADS)

    Allier, C.; Vincent, R.; Navarro, F.; Menneteau, M.; Ghenim, L.; Gidrol, X.; Bordy, T.; Hervé, L.; Cioni, O.; Bardin, S.; Bornens, M.; Usson, Y.; Morales, S.

    2018-02-01

    We present our implementation of lens-free video microscopy setup for the monitoring of adherent cell cultures. We use a multi-wavelength LED illumination together with a dedicated holographic reconstruction algorithm that allows for an efficient removal of twin images from the reconstructed phase image for densities up to those of confluent cell cultures (>500 cells/mm2). We thereby demonstrate that lens-free video microscopy, with a large field of view ( 30 mm2) can enable us to capture the images of thousands of cells simultaneously and directly inside the incubator. It is then possible to trace and quantify single cells along several cell cycles. We thus prove that lens-free microscopy is a quantitative phase imaging technique enabling estimation of several metrics at the single cell level as a function of time, for example the area, dry mass, maximum thickness, major axis length and aspect ratio of each cell. Combined with cell tracking, it is then possible to extract important parameters such as the initial cell dry mass (just after cell division), the final cell dry mass (just before cell division), the average cell growth rate, and the cell cycle duration. As an example, we discuss the monitoring of a HeLa cell cultures which provided us with a data-set featuring more than 10 000 cell cycle tracks and more than 2x106 cell morphological measurements in a single time-lapse.

  6. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e260338 - iss042e261334). Shows night time Earth views taken from the Cupola module.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    ERIC Educational Resources Information Center

    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. Quantitative phase microscopy via optimized inversion of the phase optical transfer function.

    PubMed

    Jenkins, Micah H; Gaylord, Thomas K

    2015-10-01

    Although the field of quantitative phase imaging (QPI) has wide-ranging biomedical applicability, many QPI methods are not well-suited for such applications due to their reliance on coherent illumination and specialized hardware. By contrast, methods utilizing partially coherent illumination have the potential to promote the widespread adoption of QPI due to their compatibility with microscopy, which is ubiquitous in the biomedical community. Described herein is a new defocus-based reconstruction method that utilizes a small number of efficiently sampled micrographs to optimally invert the partially coherent phase optical transfer function under assumptions of weak absorption and slowly varying phase. Simulation results are provided that compare the performance of this method with similar algorithms and demonstrate compatibility with large phase objects. The accuracy of the method is validated experimentally using a microlens array as a test phase object. Lastly, time-lapse images of live adherent cells are obtained with an off-the-shelf microscope, thus demonstrating the new method's potential for extending QPI capability widely in the biomedical community.

  10. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e196791 - iss042e197504). Shows Earth views. Day time views turn into night time views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed

    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

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

    PubMed Central

    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

  14. Time-lapse recordings of human corneal epithelial healing.

    PubMed

    Hardarson, Thorir; Hanson, Charles; Claesson, Margareta; Stenevi, Ulf

    2004-04-01

    The aim of this study was to design an experimental set-up for the study of human corneal epithelial wound healing in a controlled in vitro situation. A time-lapse set-up was used. This allowed for pictures to be captured with a magnification ranging from x 80 to x 1800. Pictures were captured at 1-min intervals during the observation period, which lasted up to 4 days. Human corneal tissue was obtained from the Eye Bank or from surgery. A small, rounded lesion was produced in the corneal epithelium with a miniature drill. The specimens were placed in a mini-incubator; the camera focused on the epithelial lesion and continuously observed using the time-lapse set-up. The healing process of human corneal epithelium could be followed for several days. The initial healing response could be divided into a slow, a rapid and a consolidating phase. The first two phases lasted about 12 hours, and by then, epithelial cells covered the lesion. Depending on the origin of the tissue and the placement of the lesion, variations in the healing response could be seen. The time-lapse technique makes it possible to study epithelial wound healing over time at the cellular level. Data collected in this way can fill the gap between in vivo studies, where, by nature, human wound healing studies are restricted, and cell culture techniques, where cellular responses in many cases differ from the in vivo situation.

  15. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e103580 - iss042e104044). Shows night time Earth views. Solar Array Wing (SAW) and Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  16. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e190769 - iss042e191096). Shows Earth views. Solar Array Wing (SAW) in foreground.

  17. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e330173 - iss042e331530). Shows Earth views. Solar Array Wing (SAW) in foreground.

  18. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e238532 - iss042e239150). Shows Earth views. Solar Array Wing (SAW) in foreground.

  19. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e177446 - iss042e178444 ). Shows Earth views. Solar Array Wing (SAW) in foreground.

  20. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e110489 - iss042e111902). Shows Earth views. Solar Array Wing (SAW) in foreground.

  1. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e212874 - iss042e213080). Shows Earth views. Solar Array Wing (SAW) in foreground.

  2. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e285752 - iss042e286830). Shows Earth views. Solar Array Wing (SAW) in foreground.

  3. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e116561 - iss042e117265). Shows Earth views. Solar Array Wing (SAW) in foreground.

  4. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (iss042e071550 - iss042e072050). Shows Earth views over Africa, Sinai, Saudi Arabia, Jordan and Israel.

  5. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e218184 - iss042e219070 ). Shows night time views over Egypt, Sinai, Saudi Arabia, Jordan and Israel. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  6. Time-lapse Inversion of Electrical Resistivity Data

    NASA Astrophysics Data System (ADS)

    Nguyen, F.; Kemna, A.

    2005-12-01

    Time-lapse geophysical measurements (also known as monitoring, repeat or multi-frame survey) now play a critical role for monitoring -non-destructively- changes induced by human, as reservoir compaction, or to study natural processes, as flow and transport in porous media. To invert such data sets into time-varying subsurface properties, several strategies are found in different engineering or scientific fields (e.g., in biomedical, process tomography, or geophysical applications). Indeed, for time-lapse surveys, the data sets and the models at each time frame have the particularity to be closely related to their "neighbors", if the process does not induce chaotic or very high variations. Therefore, the information contained in the different frames can be used for constraining the inversion in the others. A first strategy consists in imposing constraints to the model based on prior estimation, a priori spatiotemporal or temporal behavior (arbitrary or based on a law describing the monitored process), restriction of changes in certain areas, or data changes reproducibility. A second strategy aims to invert directly the model changes, where the objective function penalizes those models whose spatial, temporal, or spatiotemporal behavior differs from a prior assumption or from a computed a priori. Clearly, the incorporation of time-lapse a priori information, determined from data sets or assumed, in the inversion process has been proven to improve significantly the resolving capability, mainly by removing artifacts. However, there is a lack of comparison of these methods. In this paper, we focus on Tikhonov-like inversion approaches for electrical tomography imaging to evaluate the capability of the different existing strategies, and to propose new ones. To evaluate the bias inevitably introduced by time-lapse regularization, we quantified the relative contribution of the different approaches to the resolving power of the method. Furthermore, we incorporated different

  7. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    s time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e207712 - iss042e209132 ). Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  8. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e203119 - iss042e203971). Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  9. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e334978 - iss042e335976). Shows Earth views. Solar Array Wing (SAW) comes into view.

  10. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e324104 - iss042e325631). Shows Earth views. Soyuz and Progress spacecrafts come into view.

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

    USGS Publications Warehouse

    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.

  12. Source Repeatability of Time-Lapse Offset VSP Surveys for Monitoring CO2 Injection

    NASA Astrophysics Data System (ADS)

    Zhang, Z.; Huang, L.; Rutledge, J. T.; Denli, H.; Zhang, H.; McPherson, B. J.; Grigg, R.

    2009-12-01

    Time-lapse vertical seismic profiling (VSP) surveys have the potential to remotely track the migration of injected CO2 within a geologic formation. To accurately detect small changes due to CO2 injection, the sources of time-lapse VSP surveys must be located exactly at the same positions. However, in practice, the source locations can vary from one survey to another survey. Our numerical simulations demonstrate that a variation of a few meters in the VSP source locations can result in significant changes in time-lapse seismograms. To address the source repeatability issue, we apply double-difference tomography to downgoing waves of time-lapse offset VSP data to invert for the source locations and the velocity structures simultaneously. In collaboration with Resolute Natural Resources, Navajo National Oil and Gas Company, and the Southwest Regional Partnership on Carbon Sequestration under the support of the U.S. Department of Energy’s National Energy Technology Laboratory, one baseline and two repeat offset VSP datasets were acquired in 2007-2009 for monitoring CO2 injection at the Aneth oil field in Utah. A cemented geophone string was used to acquire the data for one zero-offset and seven offset source locations. During the data acquisition, there was some uncertainty in the repeatability of the source locations relative to the baseline survey. Our double-difference tomography results of the Aneth time-lapse VSP data show that the source locations for different surveys are separated up to a few meters. Accounting for these source location variations during VSP data analysis will improve reliability of time-lapse VSP monitoring.

  13. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e210380 - iss042e211441). Shows Earth views. Solar Array Wing (SAW) in and out of view.

  14. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e211498 - iss042e212135). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground

  15. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e162807 - iss042e163936). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  16. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e308288 - iss042e309536). Shows Earth views taken from a window aboard the International Space Station (ISS).

  17. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e193144 - iss042e194102). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  18. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e209133 - iss042e210379). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  19. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e215401 -iss042e215812). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  20. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e290689 - iss042e291289). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  1. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e249923 - iss042e250759). Shows Earth views. Space Station Remote Manipulator system (SSRMS) or Canadarm in foreground.

  2. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e170341 - iss042e171462). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  3. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2015-05-18

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss042e244330 - iss042e245101). Shows Earth views. Space Station Remote Manipulator System (SSRMS) or Canadarm in foreground.

  4. ISS Expedition 42 Time Lapse Video of Earth

    NASA Image and Video Library

    2014-09-29

    This time lapse video taken during ISS Expedition 42 is assembled from JSC still photo collection (still photos iss041e37762 - iss041e39788). Shows Earth and aurora views. Partial views of ISS in and out of view.

  5. Time-lapse joint inversion of geophysical data with automatic joint constraints and dynamic attributes

    NASA Astrophysics Data System (ADS)

    Rittgers, J. B.; Revil, A.; Mooney, M. A.; Karaoulis, M.; Wodajo, L.; Hickey, C. J.

    2016-12-01

    Joint inversion and time-lapse inversion techniques of geophysical data are often implemented in an attempt to improve imaging of complex subsurface structures and dynamic processes by minimizing negative effects of random and uncorrelated spatial and temporal noise in the data. We focus on the structural cross-gradient (SCG) approach (enforcing recovered models to exhibit similar spatial structures) in combination with time-lapse inversion constraints applied to surface-based electrical resistivity and seismic traveltime refraction data. The combination of both techniques is justified by the underlying petrophysical models. We investigate the benefits and trade-offs of SCG and time-lapse constraints. Using a synthetic case study, we show that a combined joint time-lapse inversion approach provides an overall improvement in final recovered models. Additionally, we introduce a new approach to reweighting SCG constraints based on an iteratively updated normalized ratio of model sensitivity distributions at each time-step. We refer to the new technique as the Automatic Joint Constraints (AJC) approach. The relevance of the new joint time-lapse inversion process is demonstrated on the synthetic example. Then, these approaches are applied to real time-lapse monitoring field data collected during a quarter-scale earthen embankment induced-piping failure test. The use of time-lapse joint inversion is justified by the fact that a change of porosity drives concomitant changes in seismic velocities (through its effect on the bulk and shear moduli) and resistivities (through its influence upon the formation factor). Combined with the definition of attributes (i.e. specific characteristics) of the evolving target associated with piping, our approach allows localizing the position of the preferential flow path associated with internal erosion. This is not the case using other approaches.

  6. High-Resolution Time-Lapse Monitoring of Unsaturated Flow using Automated GPR Data Collection

    NASA Astrophysics Data System (ADS)

    Mangel, A. R.; Moysey, S. M.; Lytle, B. A.; Bradford, J. H.

    2015-12-01

    High-resolution ground-penetrating radar (GPR) data provide the detailed information required to image subsurface structures. Recent advances in GPR monitoring now also make it possible to study transient hydrologic processes, but high-speed data acquisition is critical for this application. We therefore highlight the capabilities of our automated system to acquire time-lapse, high-resolution multifold GPR data during infiltration of water into soils. The system design allows for fast acquisition of constant-offset (COP) and common-midpoint profiles (CMP) to monitor unsaturated flow at multiple locations. Qualitative interpretation of the unprocessed COPs can provide substantial information regarding the hydrologic response of the system, such as the complexities of patterns associated with the wetting of the soil and geophysical evidence of non-uniform propagation of a wetting front. While we find that unprocessed images are informative, we show that the spatial variability of velocity introduced by infiltration events can complicate the images and that migration of the data is an effective tool to improve interpretability of the time-lapse images. The ability of the system to collect high density CMP data also introduces the potential for improving the velocity model along with the image via reflection tomography in the post-migrated domain. We show that for both simulated and empirical time-lapse GPR profiles we can resolve a propagating wetting front in the soil that is in good agreement with the response of in-situ soil moisture measurements. The data from these experiments illustrate the importance of high-speed, high-resolution GPR data acquisition for obtaining insight about the dynamics of hydrologic events. Continuing research is aimed at improving the quantitative analysis of surface-based GPR monitoring data for identifying preferential flow in soils.

  7. Automatic detection and analysis of cell motility in phase-contrast time-lapse images using a combination of maximally stable extremal regions and Kalman filter approaches.

    PubMed

    Kaakinen, M; Huttunen, S; Paavolainen, L; Marjomäki, V; Heikkilä, J; Eklund, L

    2014-01-01

    Phase-contrast illumination is simple and most commonly used microscopic method to observe nonstained living cells. Automatic cell segmentation and motion analysis provide tools to analyze single cell motility in large cell populations. However, the challenge is to find a sophisticated method that is sufficiently accurate to generate reliable results, robust to function under the wide range of illumination conditions encountered in phase-contrast microscopy, and also computationally light for efficient analysis of large number of cells and image frames. To develop better automatic tools for analysis of low magnification phase-contrast images in time-lapse cell migration movies, we investigated the performance of cell segmentation method that is based on the intrinsic properties of maximally stable extremal regions (MSER). MSER was found to be reliable and effective in a wide range of experimental conditions. When compared to the commonly used segmentation approaches, MSER required negligible preoptimization steps thus dramatically reducing the computation time. To analyze cell migration characteristics in time-lapse movies, the MSER-based automatic cell detection was accompanied by a Kalman filter multiobject tracker that efficiently tracked individual cells even in confluent cell populations. This allowed quantitative cell motion analysis resulting in accurate measurements of the migration magnitude and direction of individual cells, as well as characteristics of collective migration of cell groups. Our results demonstrate that MSER accompanied by temporal data association is a powerful tool for accurate and reliable analysis of the dynamic behaviour of cells in phase-contrast image sequences. These techniques tolerate varying and nonoptimal imaging conditions and due to their relatively light computational requirements they should help to resolve problems in computationally demanding and often time-consuming large-scale dynamical analysis of cultured cells.

  8. Quantitative assessment of cancer cell morphology and movement using telecentric digital holographic microscopy

    NASA Astrophysics Data System (ADS)

    Nguyen, Thanh C.; Nehmetallah, George; Lam, Van; Chung, Byung Min; Raub, Christopher

    2017-02-01

    Digital holographic microscopy (DHM) provides label-free and real-time quantitative phase information relevant to the analysis of dynamic biological systems. A DHM based on telecentric configuration optically mitigates phase aberrations due to the microscope objective and linear high frequency fringes due to the reference beam thus minimizing digital aberration correction needed for distortion free 3D reconstruction. The purpose of this work is to quantitatively assess growth and migratory behavior of invasive cancer cells using a telecentric DHM system. Together, the height and lateral shape features of individual cells, determined from time-lapse series of phase reconstructions, should reveal aspects of cell migration, cell-matrix adhesion, and cell cycle phase transitions. To test this, MDA-MB-231 breast cancer cells were cultured on collagen-coated or un-coated glass, and 3D holograms were reconstructed over 2 hours. Cells on collagencoated glass had an average 14% larger spread area than cells on uncoated glass (n=18-22 cells/group). The spread area of cells on uncoated glass were 15-21% larger than cells seeded on collagen hydrogels (n=18-22 cells/group). Premitotic cell rounding was observed with average phase height increasing 57% over 10 minutes. Following cell division phase height decreased linearly (R2=0.94) to 58% of the original height pre-division. Phase objects consistent with lamellipodia were apparent from the reconstructions at the leading edge of migrating cells. These data demonstrate the ability to track quantitative phase parameters and relate them to cell morphology during cell migration and division on adherent substrates, using telecentric DHM. The technique enables future studies of cell-matrix interactions relevant to cancer.

  9. Revealing the secret life of pre-implantation embryos by time-lapse monitoring: A review

    PubMed Central

    Faramarzi, Azita; Khalili, Mohammad Ali; Micara, Giulietta; Agha-Rahimi, Azam

    2017-01-01

    High implantation success following in vitro fertilization cycles are achieved via the transfer of embryos with the highest developmental competence. Multiple pregnancies as a result of the transfer of several embryos per cycle accompany with various complication. Thus, single-embryo transfer (SET) is the preferred practice in assisted reproductive technique (ART) treatment. In order to improve the pregnancy rate for SET, embryologists need reliable biomarkers to aid their selection of embryos with the highest developmental potential. Time-lapse technology is a noninvasive alternative conventional microscopic assessment. It provides uninterrupted and continues the survey of embryo development to transfer day. Today, there are four time-lapse systems that are commercially available for ART centers. In world and Iran, the first time lapse babies were born in 2010 and 2015, respectively, conceived by SET. Here, we review the use of time-lapse monitoring in the observation of embryogenesis as well as its role in SET. Although, the findings from our review support common use of time-lapse monitoring in ART centers; but, future large studies assessing this system in well-designed trials are necessary. PMID:28744520

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

    PubMed

    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.

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

    USGS Publications Warehouse

    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.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Singha, Kamini; Day-Lewis, Frederick D.; Johnson, Timothy C.

    2015-03-15

    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.

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

    PubMed

    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.

  14. Tracking multiple particles in fluorescence time-lapse microscopy images via probabilistic data association.

    PubMed

    Godinez, William J; Rohr, Karl

    2015-02-01

    Tracking subcellular structures as well as viral structures displayed as 'particles' in fluorescence microscopy images yields quantitative information on the underlying dynamical processes. We have developed an approach for tracking multiple fluorescent particles based on probabilistic data association. The approach combines a localization scheme that uses a bottom-up strategy based on the spot-enhancing filter as well as a top-down strategy based on an ellipsoidal sampling scheme that uses the Gaussian probability distributions computed by a Kalman filter. The localization scheme yields multiple measurements that are incorporated into the Kalman filter via a combined innovation, where the association probabilities are interpreted as weights calculated using an image likelihood. To track objects in close proximity, we compute the support of each image position relative to the neighboring objects of a tracked object and use this support to recalculate the weights. To cope with multiple motion models, we integrated the interacting multiple model algorithm. The approach has been successfully applied to synthetic 2-D and 3-D images as well as to real 2-D and 3-D microscopy images, and the performance has been quantified. In addition, the approach was successfully applied to the 2-D and 3-D image data of the recent Particle Tracking Challenge at the IEEE International Symposium on Biomedical Imaging (ISBI) 2012.

  15. Time-lapse seismic waveform inversion for monitoring near-surface microbubble injection

    NASA Astrophysics Data System (ADS)

    Kamei, R.; Jang, U.; Lumley, D. E.; Mouri, T.; Nakatsukasa, M.; Takanashi, M.

    2016-12-01

    Seismic monitoring of the Earth provides valuable information regarding the time-varying changes in subsurface physical properties that are caused by natural or man-made processes. However, the resulting changes in subsurface properties are often small both in terms of magnitude and spatial extent, leading to seismic data differences that are difficult to detect at typical non-repeatable noise levels. In order to better extract information from the time-lapse data, exploiting the full seismic waveform information can be critical, since detected amplitude or traveltime changes may be minimal. We explore methods of waveform inversion that estimate an optimal model of time-varying elastic parameters at the wavelength scale to fit the observed time-lapse seismic data with modelled waveforms based on numerical solutions of the wave equation. We apply acoustic waveform inversion to time-lapse cross-well monitoring surveys of 64-m well intervals, and estimate the velocity changes that occur during the injection of microbubble water into shallow unconsolidated Quaternary sediments in the Kanto basin of Japan at a depth of 25 m below the surface. Microbubble water is comprised of water infused with air bubbles of a diameter less than 0.1mm, and may be useful to improve resistance to ground liquefaction during major earthquakes. Monitoring the space-time distribution and physical properties of microbubble injection is therefore important to understanding the full potential of the technique. Repeated monitoring surveys (>10) reveal transient behaviours in waveforms during microbubble injection. Time-lapse waveform inversion detects changes in P-wave velocity of less than 1 percent, initially as velocity increases and subsequently as velocity decreases. The velocity changes are mainly imaged within a thin (1 m) layer between the injection and the receiver well, inferring the fluid-flow influence of the fluvial sediment depositional environment. The resulting velocity models

  16. Time-Lapse Acoustic Impedance Inversion in CO2 Sequestration Study (Weyburn Field, Canada)

    NASA Astrophysics Data System (ADS)

    Wang, Y.; Morozov, I. B.

    2016-12-01

    Acoustic-impedance (AI) pseudo-logs are useful for characterising subtle variations of fluid content during seismic monitoring of reservoirs undergoing enhanced oil recovery and/or geologic CO2 sequestration. However, highly accurate AI images are required for time-lapse analysis, which may be difficult to achieve with conventional inversion approaches. In this study, two enhancements of time-lapse AI analysis are proposed. First, a well-known uncertainty of AI inversion is caused by the lack of low-frequency signal in reflection seismic data. To resolve this difficulty, we utilize an integrated AI inversion approach combining seismic data, acoustic well logs and seismic-processing velocities. The use of well logs helps stabilizing the recursive AI inverse, and seismic-processing velocities are used to complement the low-frequency information in seismic records. To derive the low-frequency AI from seismic-processing velocity data, an empirical relation is determined by using the available acoustic logs. This method is simple and does not require subjective choices of parameters and regularization schemes as in the more sophisticated joint inversion methods. The second improvement to accurate time-lapse AI imaging consists in time-variant calibration of reflectivity. Calibration corrections consist of time shifts, amplitude corrections, spectral shaping and phase rotations. Following the calibration, average and differential reflection amplitudes are calculated, from which the average and differential AI are obtained. The approaches are applied to a time-lapse 3-D 3-C dataset from Weyburn CO2 sequestration project in southern Saskatchewan, Canada. High quality time-lapse AI volumes are obtained. Comparisons with traditional recursive and colored AI inversions (obtained without using seismic-processing velocities) show that the new method gives a better representation of spatial AI variations. Although only early stages of monitoring seismic data are available, time-lapse

  17. Localized time-lapse elastic waveform inversion using wavefield injection and extrapolation: 2-D parametric studies

    NASA Astrophysics Data System (ADS)

    Yuan, Shihao; Fuji, Nobuaki; Singh, Satish; Borisov, Dmitry

    2017-06-01

    We present a methodology to invert seismic data for a localized area by combining source-side wavefield injection and receiver-side extrapolation method. Despite the high resolving power of seismic full waveform inversion, the computational cost for practical scale elastic or viscoelastic waveform inversion remains a heavy burden. This can be much more severe for time-lapse surveys, which require real-time seismic imaging on a daily or weekly basis. Besides, changes of the structure during time-lapse surveys are likely to occur in a small area rather than the whole region of seismic experiments, such as oil and gas reservoir or CO2 injection wells. We thus propose an approach that allows to image effectively and quantitatively the localized structure changes far deep from both source and receiver arrays. In our method, we perform both forward and back propagation only inside the target region. First, we look for the equivalent source expression enclosing the region of interest by using the wavefield injection method. Second, we extrapolate wavefield from physical receivers located near the Earth's surface or on the ocean bottom to an array of virtual receivers in the subsurface by using correlation-type representation theorem. In this study, we present various 2-D elastic numerical examples of the proposed method and quantitatively evaluate errors in obtained models, in comparison to those of conventional full-model inversions. The results show that the proposed localized waveform inversion is not only efficient and robust but also accurate even under the existence of errors in both initial models and observed data.

  18. Quantitative X-ray Differential Interference Contrast Microscopy

    NASA Astrophysics Data System (ADS)

    Nakamura, Takashi

    Full-field soft x-ray microscopes are widely used in many fields of sciences. Advances in nanofabrication technology enabled short wavelength focusing elements with significantly improved spatial resolution. In the soft x-ray spectral region, samples as small as 12 nm can be resolved using micro zone-plates as the objective lens. In addition to conventional x-ray microscopy in which x-ray absorption difference provides the image contrast, phase contrast mechanisms such as differential phase contrast (DIC) and Zernike phase contrast have also been demonstrated These phase contrast imaging mechanisms are especially attractive at the x-ray wavelengths where phase contrast of most materials is typically 10 times stronger than the absorption contrast. With recent progresses in plasma-based x- ray sources and increasing accessibility to synchrotron user facilities, x-ray microscopes are quickly becoming standard measurement equipment in the laboratory. To further the usefulness of x-ray DIC microscopy this thesis explicitly addresses three known issues with this imaging modality by introducing new techniques and devices First, as opposed to its visible-light counterpart, no quantitative phase imaging technique exists for x-ray DIC microscopy. To address this issue, two nanoscale x-ray quantitative phase imaging techniques, using exclusive OR (XOR) patterns and zone-plate doublets, respectively, are proposed. Unlike existing x-ray quantitative phase imaging techniques such as Talbot interferometry and ptychography, no dedicated experimental setups or stringent illumination coherence are needed for quantitative phase retrieval. Second, to the best of our knowledge, no quantitative performance characterization of DIC microscopy exists to date. Therefore the imaging system's response to sample's spatial frequency is not known In order to gain in-depth understanding of this imaging modality, performance of x-ray DIC microscopy is quantified using modulation transfer function

  19. Unsupervised automated high throughput phenotyping of RNAi time-lapse movies.

    PubMed

    Failmezger, Henrik; Fröhlich, Holger; Tresch, Achim

    2013-10-04

    Gene perturbation experiments in combination with fluorescence time-lapse cell imaging are a powerful tool in reverse genetics. High content applications require tools for the automated processing of the large amounts of data. These tools include in general several image processing steps, the extraction of morphological descriptors, and the grouping of cells into phenotype classes according to their descriptors. This phenotyping can be applied in a supervised or an unsupervised manner. Unsupervised methods are suitable for the discovery of formerly unknown phenotypes, which are expected to occur in high-throughput RNAi time-lapse screens. We developed an unsupervised phenotyping approach based on Hidden Markov Models (HMMs) with multivariate Gaussian emissions for the detection of knockdown-specific phenotypes in RNAi time-lapse movies. The automated detection of abnormal cell morphologies allows us to assign a phenotypic fingerprint to each gene knockdown. By applying our method to the Mitocheck database, we show that a phenotypic fingerprint is indicative of a gene's function. Our fully unsupervised HMM-based phenotyping is able to automatically identify cell morphologies that are specific for a certain knockdown. Beyond the identification of genes whose knockdown affects cell morphology, phenotypic fingerprints can be used to find modules of functionally related genes.

  20. 2D Time-lapse Seismic Tomography Using An Active Time Constraint (ATC) Approach

    EPA Science Inventory

    We propose a 2D seismic time-lapse inversion approach to image the evolution of seismic velocities over time and space. The forward modeling is based on solving the eikonal equation using a second-order fast marching method. The wave-paths are represented by Fresnel volumes rathe...

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

    USGS Publications Warehouse

    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.

  2. Long Time-lapse Nanoscopy with Spontaneously Blinking Membrane Probes

    PubMed Central

    Takakura, Hideo; Zhang, Yongdeng; Erdmann, Roman S.; Thompson, Alexander D.; Lin, Yu; McNellis, Brian; Rivera-Molina, Felix; Uno, Shin-nosuke; Kamiya, Mako; Urano, Yasuteru; Rothman, James E.; Bewersdorf, Joerg; Schepartz, Alanna; Toomre, Derek

    2017-01-01

    Long time-lapse, diffraction-unlimited super-resolution imaging of cellular structures and organelles in living cells is highly challenging, as it requires dense labeling, bright, highly photostable dyes, and non-toxic conditions. We developed a set of high-density, environment-sensitive (HIDE) membrane probes based on HMSiR that assemble in situ and enable long time-lapse, live cell nanoscopy of discrete cellular structures and organelles with high spatio-temporal resolution. HIDE-enabled nanoscopy movies are up to 50x longer than movies obtained with labeled proteins, reveal the 2D dynamics of the mitochondria, plasma membrane, and filopodia, and the 2D and 3D dynamics of the endoplasmic reticulum in living cells. These new HIDE probes also facilitate the acquisition of live cell, two-color, super-resolution images, greatly expanding the utility of nanoscopy to visualize processes and structures in living cells. PMID:28671662

  3. Simultaneous Gram and viability staining on activated sludge exposed to erythromycin: 3D CLSM time-lapse imaging of bacterial disintegration.

    PubMed

    Louvet, Jean-Noël; Attik, Ghania; Dumas, Dominique; Potier, Olivier; Pons, Marie-Noëlle

    2011-11-01

    The effect of erythromycin on activated sludge bacteria according to their Gram type was investigated with 3-dimensional Confocal Laser Scanning Microscopy (CLSM) time-lapse imaging. The fluorescent stains SYTOX Green and Texas Red-X conjugate of wheat germ agglutinin stained dying bacteria and Gram(+) bacteria respectively. Time-lapse imaging allowed an understanding of the staining mechanism and the measurement of the death rate. In presence of erythromycin (10mg/L), Gram(+) bacteria had a higher mortality rate than the Gram(-) bacteria. This result suggests that antibiotic in wastewater could change the activated sludge bacteria composition, according to their Gram type by selecting the bacteria which are the least sensitive to the antibiotics. However bacterial death was followed by bacterial disintegration leading to a decrease in the fluorescence. Results suggested that the viability indicators based on membrane integrity should be used with a correct sampling method, which can give the initial quantity of living bacteria. Copyright © 2011 Elsevier GmbH. All rights reserved.

  4. Identification of biogeochemical hot spots using time-lapse hydrogeophysics

    NASA Astrophysics Data System (ADS)

    Franz, T. E.; Loecke, T.; Burgin, A.

    2016-12-01

    The identification and monitoring of biogeochemical hot spots and hot moments is difficult using point based sampling techniques and sensors. Without proper monitoring and accounting of water, energy, and trace gas fluxes it is difficult to assess the environmental footprint of land management practices. One key limitation is optimal placement of sensors/chambers that adequately capture the point scale fluxes and thus a reasonable integration to landscape scale flux. In this work we present time-lapse hydrogeophysical imaging at an old agricultural field converted into a wetland mitigation bank near Dayton, Ohio. While the wetland was previously instrumented with a network of soil sensors and surface chambers to capture a suite of state variables and fluxes, we hypothesize that time-lapse hydrogeophysical imaging is an underutilized and critical reconnaissance tool for effective network design and landscape scaling. Here we combine the time-lapse hydrogeophysical imagery with the multivariate statistical technique of Empirical Orthogonal Functions (EOF) in order to isolate the spatial and temporal components of the imagery. Comparisons of soil core information (e.g. soil texture, soil carbon) from around the study site and organized within like spatial zones reveal statistically different mean values of soil properties. Moreover, the like spatial zones can be used to identify a finite number of future sampling locations, evaluation of the placement of existing sensors/chambers, upscale/downscale observations, all of which are desirable techniques for commercial use in precision agriculture. Finally, we note that combining the EOF analysis with continuous monitoring from point sensors or remote sensing products may provide a robust statistical framework for scaling observations through time as well as provide appropriate datasets for use in landscape biogeochemical models.

  5. Monitoring Kilauea Volcano Using Non-Telemetered Time-Lapse Camera Systems

    NASA Astrophysics Data System (ADS)

    Orr, T. R.; Hoblitt, R. P.

    2006-12-01

    Systematic visual observations are an essential component of monitoring volcanic activity. At the Hawaiian Volcano Observatory, the development and deployment of a new generation of high-resolution, non- telemetered, time-lapse camera systems provides periodic visual observations in inaccessible and hazardous environments. The camera systems combine a hand-held digital camera, programmable shutter-release, and other off-the-shelf components in a package that is inexpensive, easy to deploy, and ideal for situations in which the probability of equipment loss due to volcanic activity or theft is substantial. The camera systems have proven invaluable in correlating eruptive activity with deformation and seismic data streams. For example, in late 2005 and much of 2006, Pu`u `O`o, the active vent on Kilauea Volcano`s East Rift Zone, experienced 10--20-hour cycles of inflation and deflation that correlated with increases in seismic energy release. A time-lapse camera looking into a skylight above the main lava tube about 1 km south of the vent showed an increase in lava level---an indicator of increased lava flux---during periods of deflation, and a decrease in lava level during periods of inflation. A second time-lapse camera, with a broad view of the upper part of the active flow field, allowed us to correlate the same cyclic tilt and seismicity with lava breakouts from the tube. The breakouts were accompanied by rapid uplift and subsidence of shatter rings over the tube. The shatter rings---concentric rings of broken rock---rose and subsided by as much as 6 m in less than an hour during periods of varying flux. Time-lapse imagery also permits improved assessment of volcanic hazards, and is invaluable in illustrating the hazards to the public. In collaboration with Hawaii Volcanoes National Park, camera systems have been used to monitor the growth of lava deltas at the entry point of lava into the ocean to determine the potential for catastrophic collapse.

  6. Development of factors to convert frequency to rate for β-cell replication and apoptosis quantified by time-lapse video microscopy and immunohistochemistry

    PubMed Central

    Saisho, Yoshifumi; Manesso, Erica; Gurlo, Tatyana; Huang, Chang-jiang; Toffolo, Gianna M.; Cobelli, Claudio; Butler, Peter C.

    2009-01-01

    An obstacle to development of methods to quantify β-cell turnover from pancreas tissue is the lack of conversion factors for the frequency of β-cell replication or apoptosis detected by immunohistochemistry to rates of replication or apoptosis. We addressed this obstacle in islets from 1-mo-old rats by quantifying the relationship between the rate of β-cell replication observed directly by time-lapse video microscopy (TLVM) and the frequency of β-cell replication in the same islets detected by immunohistochemistry using antibodies against Ki67 and insulin in the same islets fixed immediately after TLVM. Similarly, we quantified the rate of β-cell apoptosis by TLVM and then the frequency of apoptosis in the same islets using TdT-mediated dUTP nick-end labeling and insulin. Conversion factors were developed by regression analysis. The conversion factor from Ki67 labeling frequency (%) to actual replication rate (%events/h) is 0.025 ± 0.003 h−1. The conversion factor from TdT-mediated dUTP nick-end labeling frequency (%) to actual apoptosis rate (%events/h) is 0.41 ± 0.05 h−1. These conversion factors will permit development of models to evaluate β-cell turnover in fixed pancreas tissue. PMID:18940937

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

    NASA Astrophysics Data System (ADS)

    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

  8. Lapse time and frequency-dependent coda wave attenuation for Delhi and its surrounding regions

    NASA Astrophysics Data System (ADS)

    Das, Rabin; Mukhopadhyay, Sagarika; Singh, Ravi Kant; Baidya, Pushap R.

    2018-07-01

    Attenuation of seismic wave energy of Delhi and its surrounding regions has been estimated using coda of local earthquakes. Estimated quality factor (Qc) values are strongly dependent on frequency and lapse time. Frequency dependence of Qc has been estimated from the relationship Qc(f) = Q0fn for different lapse time window lengths. Q0 and n values vary from 73 to 453 and 0.97 to 0.63 for lapse time window lengths of 15 s to 90 s respectively. Average estimated frequency dependent relation is, Qc(f) = 135 ± 8f0.96±0.02 for the entire region for a window length of 30 s, where the average Qc value varies from 200 at 1.5 Hz to 1962 at 16 Hz. These values show that the region is seismically active and highly heterogeneous. The entire study region is divided into two sub-regions according to the geology of the area to investigate if there is a spatial variation in attenuation characteristics in this region. It is observed that at smaller lapse time both regions have similar Qc values. However, at larger lapse times the rate of increase of Qc with frequency is larger for Region 2 compared to Region 1. This is understandable, as it is closer to the tectonically more active Himalayan ranges and seismically more active compared to Region 1. The difference in variation of Qc with frequencies for the two regions is such that at larger lapse time and higher frequencies Region 2 shows higher Qc compared to Region 1. For lower frequencies the opposite situation is true. This indicates that there is a systematic variation in attenuation characteristics from the south (Region 1) to the north (Region 2) in the deeper part of the study area. This variation can be explained in terms of an increase in heat flow and a decrease in the age of the rocks from south to north.

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

    NASA Astrophysics Data System (ADS)

    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

  10. Role of interstitial branching in the development of visual corticocortical connections: a time-lapse and fixed-tissue analysis.

    PubMed

    Ruthazer, Edward S; Bachleda, Amelia R; Olavarria, Jaime F

    2010-12-15

    We combined fixed-tissue and time-lapse analyses to investigate the axonal branching phenomena underlying the development of topographically organized ipsilateral projections from area 17 to area 18a in the rat. These complementary approaches allowed us to relate static, large-scale information provided by traditional fixed-tissue analysis to highly dynamic, local, small-scale branching phenomena observed with two-photon time-lapse microscopy in acute slices of visual cortex. Our fixed-tissue data revealed that labeled area 17 fibers invaded area 18a gray matter at topographically restricted sites, reaching superficial layers in significant numbers by postnatal day 6 (P6). Moreover, most parental axons gave rise to only one or occasionally a small number of closely spaced interstitial branches beneath 18a. Our time-lapse data showed that many filopodium-like branches emerged along parental axons in white matter or deep layers in area 18a. Most of these filopodial branches were transient, often disappearing after several minutes to hours of exploratory extension and retraction. These dynamic behaviors decreased significantly from P4, when the projection is first forming, through the second postnatal week, suggesting that the expression of, or sensitivity to, cortical cues promoting new branch addition in the white matter is developmentally down-regulated coincident with gray matter innervation. Together, these data demonstrate that the development of topographically organized corticocortical projections in rats involves extensive exploratory branching along parental axons and invasion of cortex by only a small number of interstitial branches, rather than the widespread innervation of superficial cortical layers by an initially exuberant population of branches. © 2010 Wiley-Liss, Inc.

  11. Role of Interstitial Branching in the Development of Visual Corticocortical Connections: A Time-Lapse and Fixed-Tissue Analysis

    PubMed Central

    Ruthazer, Edward S.; Bachleda, Amelia R.; Olavarria, Jaime F.

    2013-01-01

    We combined fixed-tissue and time-lapse analyses to investigate the axonal branching phenomena underlying the development of topographically organized ipsilateral projections from area 17 to area 18a in the rat. These complementary approaches allowed us to relate static, large-scale information provided by traditional fixed-tissue analysis to highly dynamic, local, small-scale branching phenomena observed with two-photon time-lapse microscopy in acute slices of visual cortex. Our fixed-tissue data revealed that labeled area 17 fibers invaded area 18a gray matter at topographically restricted sites, reaching superficial layers in significant numbers by postnatal day 6 (P6). Moreover, most parental axons gave rise to only one or occasionally a small number of closely spaced interstitial branches beneath 18a. Our time-lapse data showed that many filopodium-like branches emerged along parental axons in white matter or deep layers in area 18a. Most of these filopo-dial branches were transient, often disappearing after several minutes to hours of exploratory extension and retraction. These dynamic behaviors decreased significantly from P4, when the projection is first forming, through the second postnatal week, suggesting that the expression of, or sensitivity to, cortical cues promoting new branch addition in the white matter is developmentally down-regulated coincident with gray matter innervation. Together, these data demonstrate that the development of topographically organized corticocortical projections in rats involves extensive exploratory branching along parental axons and invasion of cortex by only a small number of interstitial branches, rather than the widespread innervation of superficial cortical layers by an initially exuberant population of branches. PMID:21031561

  12. Watching intracellular lipolysis in mycobacteria using time lapse fluorescence microscopy.

    PubMed

    Dhouib, Rabeb; Ducret, Adrien; Hubert, Pierre; Carrière, Frédéric; Dukan, Sam; Canaan, Stéphane

    2011-04-01

    The fact that Mycobacterium tuberculosis mobilizes lipid bodies (LB) located in the cytosol during infection process has been proposed for decades. However, the mechanisms and dynamics of mobilization of these lipid droplets within mycobacteria are still not completely characterized. Evidence in favour of this characterization was obtained here using a combined fluorescent microscopy and computational image processing approach. The decrease in lipid storage levels observed under nutrient depletion conditions was correlated with a significant increase in the size of the bacteria. LB fragmentation/condensation cycles were monitored in real time. The exact contribution of lipases in this process was confirmed using the lipase inhibitor tetrahydrolipstatin, which was found to prevent LB degradation and to limit the bacterial cell growth. The method presented here provides a powerful tool for monitoring in vivo lipolysis in mycobacteria and for obtaining new insights on the growth of cells and their entry into the dormant or reactivation phase. It should be particularly useful for studying the effects of chemical inhibitors and activators on cells as well as investigating other metabolic pathways. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. Bacterial trajectories tracked with time-lapse video-microscopy reveal the impact of manganese biomineralization on bacterial sedimentation.

    NASA Astrophysics Data System (ADS)

    Kerboas, Camille; Pena, Jasquelin; De Anna, Pietro

    2017-04-01

    In aquatic and subsurface environments, sedimentation may influence bacterial transport. Microorganisms that carry out biomineralization reactions may exhibit distinct transport properties from non-biomineralizing organisms due an apparent increase in density caused by biomineral production. For several decades, the biomineralization of manganese (Mn) has been recognized to be a major environmental process, whereby Mn oxide (MnO2(s)) minerals participate in a plethora of biogeochemical processes including contaminant adsorption, organic matter oxidation. Typically, manganese biomineralization proceeds through the enzymatic oxidation of aqueous Mn2+ to Mn4+ and precipitation of MnO2(s) in a biofilm matrix outside the bacterial cell. Here, we present a study of the impact of biomineralization on the sedimentation properties of bacteria at small scales (over mm distances) under hydrostatic conditions. We hypothesize that bacteria will sediment faster when biomineralization is active due to encrustation of the organisms by mineral particles. To test this hypothesis, we tracked the vertical notion of individual bacteria (Pseudomonas putida GB-1) using time-lapse video-microscopy. We compared the sedimentation velocity of bacteria in the case where significant biomineralization had occurred, as inferred from bulk measurements of solid phase Mn, with the sinking velocity of bacteria grown without Mn. We calibrated the proposed method by comparing velocity measurements of sinking polystyrene micro-sphere of known density and size with Stokes law, obtaining results that were accurate within 1% of the theoretical value (29.4 nm/s). We also measured a diffusion coefficient of 7x10-13 m2/s for the particles. Following this approach, we measured the sedimentation velocity of P. putida with and without MnO2(s). Our results show that biomineralization leads to faster sedimentation of the bacteria. In natural environments, biomineralization reactions may increase the sinking

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

    ERIC Educational Resources Information Center

    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…

  15. Quantitative 4D analyses of epithelial folding during Drosophila gastrulation.

    PubMed

    Khan, Zia; Wang, Yu-Chiun; Wieschaus, Eric F; Kaschube, Matthias

    2014-07-01

    Understanding the cellular and mechanical processes that underlie the shape changes of individual cells and their collective behaviors in a tissue during dynamic and complex morphogenetic events is currently one of the major frontiers in developmental biology. The advent of high-speed time-lapse microscopy and its use in monitoring the cellular events in fluorescently labeled developing organisms demonstrate tremendous promise in establishing detailed descriptions of these events and could potentially provide a foundation for subsequent hypothesis-driven research strategies. However, obtaining quantitative measurements of dynamic shapes and behaviors of cells and tissues in a rapidly developing metazoan embryo using time-lapse 3D microscopy remains technically challenging, with the main hurdle being the shortage of robust imaging processing and analysis tools. We have developed EDGE4D, a software tool for segmenting and tracking membrane-labeled cells using multi-photon microscopy data. Our results demonstrate that EDGE4D enables quantification of the dynamics of cell shape changes, cell interfaces and neighbor relations at single-cell resolution during a complex epithelial folding event in the early Drosophila embryo. We expect this tool to be broadly useful for the analysis of epithelial cell geometries and movements in a wide variety of developmental contexts. © 2014. Published by The Company of Biologists Ltd.

  16. Quantitative fluorescence microscopy and image deconvolution.

    PubMed

    Swedlow, Jason R

    2013-01-01

    Quantitative imaging and image deconvolution have become standard techniques for the modern cell biologist because they can form the basis of an increasing number of assays for molecular function in a cellular context. There are two major types of deconvolution approaches--deblurring and restoration algorithms. Deblurring algorithms remove blur but treat a series of optical sections as individual two-dimensional entities and therefore sometimes mishandle blurred light. Restoration algorithms determine an object that, when convolved with the point-spread function of the microscope, could produce the image data. The advantages and disadvantages of these methods are discussed in this chapter. Image deconvolution in fluorescence microscopy has usually been applied to high-resolution imaging to improve contrast and thus detect small, dim objects that might otherwise be obscured. Their proper use demands some consideration of the imaging hardware, the acquisition process, fundamental aspects of photon detection, and image processing. This can prove daunting for some cell biologists, but the power of these techniques has been proven many times in the works cited in the chapter and elsewhere. Their usage is now well defined, so they can be incorporated into the capabilities of most laboratories. A major application of fluorescence microscopy is the quantitative measurement of the localization, dynamics, and interactions of cellular factors. The introduction of green fluorescent protein and its spectral variants has led to a significant increase in the use of fluorescence microscopy as a quantitative assay system. For quantitative imaging assays, it is critical to consider the nature of the image-acquisition system and to validate its response to known standards. Any image-processing algorithms used before quantitative analysis should preserve the relative signal levels in different parts of the image. A very common image-processing algorithm, image deconvolution, is used

  17. Study of time-lapse processing for dynamic hydrologic conditions. [electronic satellite image analysis console for Earth Resources Technology Satellites imagery

    NASA Technical Reports Server (NTRS)

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

    1974-01-01

    The usefulness of dynamic display techniques in exploiting the repetitive nature of ERTS imagery was investigated. A specially designed Electronic Satellite Image Analysis Console (ESIAC) was developed and employed to process data for seven ERTS principal investigators studying dynamic hydrological conditions for diverse applications. These applications include measurement of snowfield extent and sediment plumes from estuary discharge, Playa Lake inventory, and monitoring of phreatophyte and other vegetation changes. 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 most unique feature of the system is the capability to time lapse the imagery and analytic displays of the imagery. Data products included quantitative measurements of distances and areas, binary thematic maps based on monospectral or multispectral decisions, radiance profiles, and movie loops. Applications of animation for uses other than creating time-lapse sequences are identified. Input to the ESIAC can be either digital or via photographic transparencies.

  18. Light sheet microscopy.

    PubMed

    Weber, Michael; Mickoleit, Michaela; Huisken, Jan

    2014-01-01

    This chapter introduces the concept of light sheet microscopy along with practical advice on how to design and build such an instrument. Selective plane illumination microscopy is presented as an alternative to confocal microscopy due to several superior features such as high-speed full-frame acquisition, minimal phototoxicity, and multiview sample rotation. Based on our experience over the last 10 years, we summarize the key concepts in light sheet microscopy, typical implementations, and successful applications. In particular, sample mounting for long time-lapse imaging and the resulting challenges in data processing are discussed in detail. © 2014 Elsevier Inc. All rights reserved.

  19. Time-lapse joint AVO inversion using generalized linear method based on exact Zoeppritz equations

    NASA Astrophysics Data System (ADS)

    Zhi, L.; Gu, H.

    2017-12-01

    The conventional method of time-lapse AVO (Amplitude Versus Offset) inversion is mainly based on the approximate expression of Zoeppritz equations. Though the approximate expression is concise and convenient to use, it has certain limitations. For example, its application condition is that the difference of elastic parameters between the upper medium and lower medium is little and the incident angle is small. In addition, the inversion of density is not stable. Therefore, we develop the method of time-lapse joint AVO inversion based on exact Zoeppritz equations. In this method, we apply exact Zoeppritz equations to calculate the reflection coefficient of PP wave. And in the construction of objective function for inversion, we use Taylor expansion to linearize the inversion problem. Through the joint AVO inversion of seismic data in baseline survey and monitor survey, we can obtain P-wave velocity, S-wave velocity, density in baseline survey and their time-lapse changes simultaneously. We can also estimate the oil saturation change according to inversion results. Compared with the time-lapse difference inversion, the joint inversion has a better applicability. It doesn't need some assumptions and can estimate more parameters simultaneously. Meanwhile, by using the generalized linear method, the inversion is easily realized and its calculation amount is small. We use the Marmousi model to generate synthetic seismic records to test and analyze the influence of random noise. Without noise, all estimation results are relatively accurate. With the increase of noise, P-wave velocity change and oil saturation change are stable and less affected by noise. S-wave velocity change is most affected by noise. Finally we use the actual field data of time-lapse seismic prospecting to process and the results can prove the availability and feasibility of our method in actual situation.

  20. Efficiency of time-lapse intervals and simple baits for camera surveys of wild pigs

    USGS Publications Warehouse

    Williams, B.L.; Holtfreter, R.W.; Ditchkoff, S.S.; Grand, J.B.

    2011-01-01

    Growing concerns surrounding established and expanding populations of wild pigs (Sus scrofa) have created the need for rapid and accurate surveys of these populations. We conducted surveys of a portion of the wild pig population on Fort Benning, Georgia, to determine if a longer time-lapse interval than had been previously used in surveys of wild pigs would generate similar detection results. We concurrently examined whether use of soured corn at camera sites affected the time necessary for pigs to locate a new camera site or the time pigs remained at a site. Our results suggest that a 9-min time-lapse interval generated dependable detection results for pigs and that soured corn neither attracted pigs to a site any quicker than plain, dry, whole-kernel corn, nor held them at a site longer. Maximization of time-lapse interval should decrease data and processing loads, and use of a simple, available bait should decrease cost and effort associated with more complicated baits; combination of these concepts should increase efficiency of wild pig surveys. ?? 2011 The Wildlife Society.

  1. Neural network control of focal position during time-lapse microscopy of cells.

    PubMed

    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.

  2. Contribution of 3-D time-lapse ERT to the study of leachate recirculation in a landfill

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Clement, R., E-mail: remi.clement@hmg.inpg.fr; Grenoble Universite, B.P. 53, 38041 Grenoble Cedex 9; Oxarango, L.

    2011-03-15

    Leachate recirculation is a key process in the operation of municipal waste landfills as bioreactors. It aims at increasing the moisture content to optimise the biodegradation. Because waste is a very heterogeneous and anisotropic porous media, the geometry of the leachate plume recirculation is difficult to delineate from the surface at the scale of the bioreactor site. In this study, 3-D time-lapse electrical resistivity tomography (ERT) was used to obtain useful information for understanding leachate recirculation hydrodynamics. The ERT inversion methodology and the electrode arrays were optimised using numerical modelling simulating a 3-D leachate injection scenario. Time-lapse ERT was subsequentlymore » applied at the field scale during an experimental injection. We compared ERT images with injected volumes to evaluate the sensitivity of time-lapse ERT to delineate the plume migration. The results show that time-lapse ERT can accomplish the following: (i) accurately locate the injection plume, delineating its depth and lateral extension; (ii) be used to estimate some hydraulic properties of waste.« less

  3. Quantitative phase-contrast digital holographic microscopy for cell dynamic evaluation

    NASA Astrophysics Data System (ADS)

    Yu, Lingfeng; Mohanty, Samarendra; Berns, Michael W.; Chen, Zhongping

    2009-02-01

    The laser microbeam uses lasers to alter and/or to ablate intracellular organelles and cellular and tissue samples, and, today, has become an important tool for cell biologists to study the molecular mechanism of complex biological systems by removing individual cells or sub-cellular organelles. However, absolute quantitation of the localized alteration/damage to transparent phase objects, such as the cell membrane or chromosomes, was not possible using conventional phase-contrast or differential interference contrast microscopy. We report the development of phase-contrast digital holographic microscopy for quantitative evaluation of cell dynamic changes in real time during laser microsurgery. Quantitative phase images are recorded during the process of laser microsurgery and thus, the dynamic change in phase can be continuously evaluated. Out-of-focus organelles are re-focused by numerical reconstruction algorithms.

  4. Heart rate measurement based on a time-lapse image.

    PubMed

    Takano, Chihiro; Ohta, Yuji

    2007-10-01

    Using a time-lapse image acquired from a CCD camera, we developed a non-contact and non-invasive device, which could measure both the respiratory and pulse rate simultaneously. The time-lapse image of a part of the subject's skin was consecutively captured, and the changes in the average image brightness of the region of interest (ROI) were measured for 30s. The brightness data were processed by a series of operations of interpolation as follows a first-order derivative, a low pass filter of 2 Hz, and a sixth-order auto-regressive (AR) spectral analysis. Fourteen sound and healthy female subjects (22-27 years of age) participated in the experiments. Each subject was told to keep a relaxed seating posture with no physical restriction. At the same time, heart rate was measured by a pulse oximeter and respiratory rate was measured by a thermistor placed at the external naris. Using AR spectral analysis, two clear peaks could be detected at approximately 0.3 and 1.2 Hz. The peaks were thought to correspond to the respiratory rate and the heart rate. Correlation coefficients of 0.90 and 0.93 were obtained for the measurement of heart rate and respiratory rate, respectively.

  5. Very-high-resolution time-lapse photography for plant and ecosystems research.

    PubMed

    Nichols, Mary H; Steven, Janet C; Sargent, Randy; Dille, Paul; Schapiro, Joshua

    2013-09-01

    Traditional photography is a compromise between image detail and area covered. We report a new method for creating time-lapse sequences of very-high-resolution photographs to produce zoomable images that facilitate observation across a range of spatial and temporal scales. • A robotic camera mount and software were used to capture images of the growth and movement in Brassica rapa every 15 s in the laboratory. The resultant time-lapse sequence (http://timemachine.gigapan.org/wiki/Plant_Growth) captures growth detail such as circumnutation. A modified, solar-powered system was deployed at a remote field site in southern Arizona. Images were collected every 2 h over a 3-mo period to capture the response of vegetation to monsoon season rainfall (http://timemachine.gigapan.org/wiki/Arizona_Grasslands). • A technique for observing time sequences of both individual plant and ecosystem response at a range of spatial scales is available for use in the laboratory and in the field.

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

    PubMed

    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.

  7. An ecological momentary intervention for smoking cessation: The associations of just-in-time, tailored messages with lapse risk factors.

    PubMed

    Hébert, Emily T; Stevens, Elise M; Frank, Summer G; Kendzor, Darla E; Wetter, David W; Zvolensky, Michael J; Buckner, Julia D; Businelle, Michael S

    2018-03-01

    Smartphone apps can provide real-time, tailored interventions for smoking cessation. The current study examines the effectiveness of a smartphone-based smoking cessation application that assessed risk for imminent smoking lapse multiple times per day and provided messages tailored to current smoking lapse risk and specific lapse triggers. Participants (N=59) recruited from a safety-net hospital smoking cessation clinic completed phone-based ecological momentary assessments (EMAs) 5 times/day for 3 consecutive weeks (1week pre-quit, 2weeks post-quit). Risk for smoking lapse was estimated in real-time using a novel weighted lapse risk estimator. With each EMA, participants received messages tailored to current level of risk for imminent smoking lapse and self-reported presence of smoking urge, stress, cigarette availability, and motivation to quit. Generalized linear mixed model analyses determined whether messages tailored to specific lapse risk factors were associated with greater reductions in these triggers than messages not tailored to specific triggers. Overall, messages tailored to smoking urge, cigarette availability, or stress corresponded with greater reductions in those triggers than messages that were not tailored to specific triggers (p's=0.02 to <0.001). Although messages tailored to stress were associated with greater reductions in stress than messages not tailored to stress, the association was non-significant (p=0.892) when only moments of high stress were included in the analysis. Mobile technology can be used to conduct real-time smoking lapse risk assessment and provide tailored treatment content. Findings provide initial evidence that tailored content may impact users' urge to smoke, stress, and cigarette availability. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Super-resolution Time-Lapse Seismic Waveform Inversion

    NASA Astrophysics Data System (ADS)

    Ovcharenko, O.; Kazei, V.; Peter, D. B.; Alkhalifah, T.

    2017-12-01

    Time-lapse seismic waveform inversion is a technique, which allows tracking changes in the reservoirs over time. Such monitoring is relatively computationally extensive and therefore it is barely feasible to perform it on-the-fly. Most of the expenses are related to numerous FWI iterations at high temporal frequencies, which is inevitable since the low-frequency components can not resolve fine scale features of a velocity model. Inverted velocity changes are also blurred when there is noise in the data, so the problem of low-resolution images is widely known. One of the problems intensively tackled by computer vision research community is the recovering of high-resolution images having their low-resolution versions. Usage of artificial neural networks to reach super-resolution from a single downsampled image is one of the leading solutions for this problem. Each pixel of the upscaled image is affected by all the pixels of its low-resolution version, which enables the workflow to recover features that are likely to occur in the corresponding environment. In the present work, we adopt machine learning image enhancement technique to improve the resolution of time-lapse full-waveform inversion. We first invert the baseline model with conventional FWI. Then we run a few iterations of FWI on a set of the monitoring data to find desired model changes. These changes are blurred and we enhance their resolution by using a deep neural network. The network is trained to map low-resolution model updates predicted by FWI into the real perturbations of the baseline model. For supervised training of the network we generate a set of random perturbations in the baseline model and perform FWI on the noisy data from the perturbed models. We test the approach on a realistic perturbation of Marmousi II model and demonstrate that it outperforms conventional convolution-based deblurring techniques.

  9. Time-resolved imaging refractometry of microbicidal films using quantitative phase microscopy.

    PubMed

    Rinehart, Matthew T; Drake, Tyler K; Robles, Francisco E; Rohan, Lisa C; Katz, David; Wax, Adam

    2011-12-01

    Quantitative phase microscopy is applied to image temporal changes in the refractive index (RI) distributions of solutions created by microbicidal films undergoing hydration. We present a novel method of using an engineered polydimethylsiloxane structure as a static phase reference to facilitate calibration of the absolute RI across the entire field. We present a study of dynamic structural changes in microbicidal films during hydration and subsequent dissolution. With assumptions about the smoothness of the phase changes induced by these films, we calculate absolute changes in the percentage of film in regions across the field of view.

  10. Improved Time-Lapsed Angular Scattering Microscopy of Single Cells

    NASA Astrophysics Data System (ADS)

    Cannaday, Ashley E.

    By measuring angular scattering patterns from biological samples and fitting them with a Mie theory model, one can estimate the organelle size distribution within many cells. Quantitative organelle sizing of ensembles of cells using this method has been well established. Our goal is to develop the methodology to extend this approach to the single cell level, measuring the angular scattering at multiple time points and estimating the non-nuclear organelle size distribution parameters. The diameters of individual organelle-size beads were successfully extracted using scattering measurements with a minimum deflection angle of 20 degrees. However, the accuracy of size estimates can be limited by the angular range detected. In particular, simulations by our group suggest that, for cell organelle populations with a broader size distribution, the accuracy of size prediction improves substantially if the minimum angle of detection angle is 15 degrees or less. The system was therefore modified to collect scattering angles down to 10 degrees. To confirm experimentally that size predictions will become more stable when lower scattering angles are detected, initial validations were performed on individual polystyrene beads ranging in diameter from 1 to 5 microns. We found that the lower minimum angle enabled the width of this delta-function size distribution to be predicted more accurately. Scattering patterns were then acquired and analyzed from single mouse squamous cell carcinoma cells at multiple time points. The scattering patterns exhibit angular dependencies that look unlike those of any single sphere size, but are well-fit by a broad distribution of sizes, as expected. To determine the fluctuation level in the estimated size distribution due to measurement imperfections alone, formaldehyde-fixed cells were measured. Subsequent measurements on live (non-fixed) cells revealed an order of magnitude greater fluctuation in the estimated sizes compared to fixed cells. With

  11. A multimethod Global Sensitivity Analysis to aid the calibration of geomechanical models via time-lapse seismic data

    NASA Astrophysics Data System (ADS)

    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

  12. Detecting cells in time varying intensity images in confocal microscopy for gene expression studies in living cells

    NASA Astrophysics Data System (ADS)

    Mitra, Debasis; Boutchko, Rostyslav; Ray, Judhajeet; Nilsen-Hamilton, Marit

    2015-03-01

    In this work we present a time-lapsed confocal microscopy image analysis technique for an automated gene expression study of multiple single living cells. Fluorescence Resonance Energy Transfer (FRET) is a technology by which molecule-to-molecule interactions are visualized. We analyzed a dynamic series of ~102 images obtained using confocal microscopy of fluorescence in yeast cells containing RNA reporters that give a FRET signal when the gene promoter is activated. For each time frame, separate images are available for three spectral channels and the integrated intensity snapshot of the system. A large number of time-lapsed frames must be analyzed to identify each cell individually across time and space, as it is moving in and out of the focal plane of the microscope. This makes it a difficult image processing problem. We have proposed an algorithm here, based on scale-space technique, which solves the problem satisfactorily. The algorithm has multiple directions for even further improvement. The ability to rapidly measure changes in gene expression simultaneously in many cells in a population will open the opportunity for real-time studies of the heterogeneity of genetic response in a living cell population and the interactions between cells that occur in a mixed population, such as the ones found in the organs and tissues of multicellular organisms.

  13. Time-lapse camera studies of sea-disposed chemical munitions in Hawaii

    NASA Astrophysics Data System (ADS)

    Edwards, Margo H.; Fornari, Daniel J.; Rognstad, Mark R.; Kelley, Christopher D.; Mah, Christopher L.; Davis, Logan K.; Flores, Kyle R. M.; Main, Erin L.; Bruso, Natalie L.

    2016-06-01

    The interactions between fauna and sea-disposed munitions provide important evidence regarding whether munitions constituents affect the health of the ocean environment and its inhabitants. To date few studies of these interactions have been conducted at deep-water disposal sites; typically observations of fauna in the vicinity of sea-disposed munitions are limited to the few minutes or hours required to collect physical samples at a specific location. During the 2012 Hawaii Undersea Military Munitions Assessment (HUMMA) field program we deployed two deep-sea time-lapse camera systems with the objectives of cataloging the diversity of fauna visiting sea-disposed chemical munitions and observing faunal behavior and physiology. Over the 1- and 3-day deployments we recorded 28 different species of fishes, crustaceans, mollusks, cnidarians, and echinoderms at the two sites. Both cameras captured the previously undocumented behavior of brisingid sea stars repositioning themselves along chemical munitions casings. Despite the fact that brisingid sea stars are able to move, for the duration of both time-lapse experiments they remained on chemical munitions casings. We interpret this result to indicate that the advantages of residing on a hard substrate slightly elevated above the seafloor outweigh the effects of chemical munitions constituents for brisingid sea stars. One type of physiological anomaly observed on several arms of the brisingid sea stars at the time-lapse sites led to the collection and examination of six specimens. As reported by Mah (2015. Deep Sea Res. II, 2015, XX-XX), these physiological features are the result of parasitic crustaceans and are not caused by chemical munitions constituents.

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

    NASA Astrophysics Data System (ADS)

    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

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

    NASA Astrophysics Data System (ADS)

    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.

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

    NASA Astrophysics Data System (ADS)

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

    2017-04-01

    Structure from Motion (SfM) photogrammetry has become an important tool for studying earth surface processes because of its flexibility, ease of use, low cost and its capability of producing high quality 3-D surface models. A major benefit of SfM is that model accuracy is fit for purpose and surveys can be designed to meet a large range of spatial and temporal scales. In the Earth sciences, research in time-lapse SfM photogrammetry or videogrammetry is an area that is difficult to undertake due to complexities in acquiring, processing and managing large 4D datasets and represents an area with significant advancement potential (Eltner et al. 2016). In this study, we investigate the potential of 4D time-lapse SfM to monitor unstable rock slopes. We tested an array of statically mounted cameras collecting time-lapse photos of a limestone rock slope located along a highway in Canada. Our setup consisted of 8 DSLR cameras with 50 mm prime lenses spaced 2-3 m apart at a distance of 10 m from the slope. The portion of the rock slope monitored was 20 m wide and 6 m high. We collected data in four phases, each having 50 photographs taken simultaneously by each camera. The first phase of photographs was taken of the stable slope. In each successive phase, we gradually moved small, discrete blocks within the rock slope by 5-15 mm, simulating pre-failure deformation of rockfall. During the last phase we also removed discrete rock blocks, simulating rockfall. We used Agisoft Photoscan's 4D processing functionality and timeline tools to create 3D point clouds from the time-lapse photographs. These tools have the benefit of attaining better accuracy photo alignments as a greater number of photos are used. For change detection, we used the 4D filtering and calibration technique proposed by Kromer et al. (2015), which takes advantage of high degrees of spatial and temporal point redundancy to decrease measurement uncertainty. Preliminary results show that it is possible to attain

  17. Time-resolved imaging refractometry of microbicidal films using quantitative phase microscopy

    PubMed Central

    Rinehart, Matthew T.; Drake, Tyler K.; Robles, Francisco E.; Rohan, Lisa C.; Katz, David; Wax, Adam

    2011-01-01

    Quantitative phase microscopy is applied to image temporal changes in the refractive index (RI) distributions of solutions created by microbicidal films undergoing hydration. We present a novel method of using an engineered polydimethylsiloxane structure as a static phase reference to facilitate calibration of the absolute RI across the entire field. We present a study of dynamic structural changes in microbicidal films during hydration and subsequent dissolution. With assumptions about the smoothness of the phase changes induced by these films, we calculate absolute changes in the percentage of film in regions across the field of view. PMID:22191912

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

    NASA Astrophysics Data System (ADS)

    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

  19. Time-lapse monitoring of zona pellucida-free embryos obtained through in vitro fertilization: a retrospective case series.

    PubMed

    Bodri, Daniel; Kato, Ryutaro; Kondo, Masae; Hosomi, Naoko; Katsumata, Yoshinari; Kawachiya, Satoshi; Matsumoto, Tsunekazu

    2015-05-01

    To report time-lapse monitoring of human oocytes in which the damaged zona pellucida was removed, producing zona-free (ZF) oocytes that were cultured until the blastocyst stage in time-lapse incubators. Retrospective case series. Private infertility clinic. Infertile patients (n = 32) undergoing minimal ovarian stimulation or natural cycle IVF treatment between October 2012 and June 2014. Intracytoplasmic sperm injection (ICSI) fertilization of ZF oocytes, prolonged embryo culture in time-lapse incubators, elective vitrification, and subsequent single vitrified-thawed blastocyst transfer (SVBT). Rate of fertilization, cleavage and blastocyst development, live-birth rate per SVBT cycle. In spite of advanced maternal age (39 ± 4.2; range, 30-46 years), good fertilization (94%), cleavage (94%), and blastocyst development rates (38%) were reached after fertilization and culturing of ZF oocytes/embryos. All thawed ZF blastocysts survived, and up to this date seven SVBT transfers were performed, yielding three (43%) term live births with healthy newborns. Time-lapse imagery gives a unique insight into the dynamics of embryo development in ZF embryos. Moreover, our case series demonstrate that an oocyte with a damaged zona pellucida that has been removed could be successfully fertilized with ICSI, cultured until blastocyst stage in a time-lapse incubator and vitrified electively for subsequent use. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. Correlative atomic force microscopy quantitative imaging-laser scanning confocal microscopy quantifies the impact of stressors on live cells in real-time.

    PubMed

    Bhat, Supriya V; Sultana, Taranum; Körnig, André; McGrath, Seamus; Shahina, Zinnat; Dahms, Tanya E S

    2018-05-29

    There is an urgent need to assess the effect of anthropogenic chemicals on model cells prior to their release, helping to predict their potential impact on the environment and human health. Laser scanning confocal microscopy (LSCM) and atomic force microscopy (AFM) have each provided an abundance of information on cell physiology. In addition to determining surface architecture, AFM in quantitative imaging (QI) mode probes surface biochemistry and cellular mechanics using minimal applied force, while LSCM offers a window into the cell for imaging fluorescently tagged macromolecules. Correlative AFM-LSCM produces complimentary information on different cellular characteristics for a comprehensive picture of cellular behaviour. We present a correlative AFM-QI-LSCM assay for the simultaneous real-time imaging of living cells in situ, producing multiplexed data on cell morphology and mechanics, surface adhesion and ultrastructure, and real-time localization of multiple fluorescently tagged macromolecules. To demonstrate the broad applicability of this method for disparate cell types, we show altered surface properties, internal molecular arrangement and oxidative stress in model bacterial, fungal and human cells exposed to 2,4-dichlorophenoxyacetic acid. AFM-QI-LSCM is broadly applicable to a variety of cell types and can be used to assess the impact of any multitude of contaminants, alone or in combination.

  1. Time-lapse joint AVO inversion using generalized linear method based on exact Zoeppritz equations

    NASA Astrophysics Data System (ADS)

    Zhi, Longxiao; Gu, Hanming

    2018-03-01

    The conventional method of time-lapse AVO (Amplitude Versus Offset) inversion is mainly based on the approximate expression of Zoeppritz equations. Though the approximate expression is concise and convenient to use, it has certain limitations. For example, its application condition is that the difference of elastic parameters between the upper medium and lower medium is little and the incident angle is small. In addition, the inversion of density is not stable. Therefore, we develop the method of time-lapse joint AVO inversion based on exact Zoeppritz equations. In this method, we apply exact Zoeppritz equations to calculate the reflection coefficient of PP wave. And in the construction of objective function for inversion, we use Taylor series expansion to linearize the inversion problem. Through the joint AVO inversion of seismic data in baseline survey and monitor survey, we can obtain the P-wave velocity, S-wave velocity, density in baseline survey and their time-lapse changes simultaneously. We can also estimate the oil saturation change according to inversion results. Compared with the time-lapse difference inversion, the joint inversion doesn't need certain assumptions and can estimate more parameters simultaneously. It has a better applicability. Meanwhile, by using the generalized linear method, the inversion is easily implemented and its calculation cost is small. We use the theoretical model to generate synthetic seismic records to test and analyze the influence of random noise. The results can prove the availability and anti-noise-interference ability of our method. We also apply the inversion to actual field data and prove the feasibility of our method in actual situation.

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

    NASA Astrophysics Data System (ADS)

    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.

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

    USGS Publications Warehouse

    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.

  4. Photographer : JPL Range : 1 million kilometers Voyager 2 completed a dramatic 10 hour time lapse

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Photographer : JPL Range : 1 million kilometers Voyager 2 completed a dramatic 10 hour time lapse photo sequence to monitor the active volcanos on Jupiter's moon Io following the spacecraft's closest approach to Jupiter. This picture is one of about 200 images that will be used to generate a time lapse motion picture to illustrate Io's volcanic activity. On the bright limb, two of the plumes (P-5 & P-6) discovered in March by Voyager 1 are again visible. The plumes are spewing materials to a height of about 100 kilometers.

  5. Time-lapse CO2 monitoring using ambient-noise seismic interferometry: a feasibility study from Ketzin, Germany

    NASA Astrophysics Data System (ADS)

    Boullenger, Boris; Verdel, Arie; Paap, Bob; Thorbecke, Jan; Draganov, Deyan

    2015-04-01

    Seismic interferometry applied to ambient-noise measurements allows retrieval of the Green's function between two seismic receivers, by cross-correlating their recordings, as if from a source at one of the receivers. We propose to use ambient-noise seismic interferometry (ANSI) to retrieve reflection data. The time-lapse differences between different vintages of the retrieved data may help characterize property changes within a geologic reservoir with varying CO2 saturation. We test the feasibility of this time-lapse passive seismic method with numerical experiments based on the CO2-storage site of Ketzin, Germany. Ambient-noise recordings from Ketzin exhibit significant passive body-wave energy (from natural tremors or induced seismicity in the vicinity of the reservoir), which is advantageous to retrieve reflections with ANSI. The ANSI numerical experiments aim to understand what the requirements are for the recorded body-wave noise to retrieve the time-lapse reflection signal caused by an increase of CO2 saturation in the reservoir. For this purpose, we design two velocity scenarios at Ketzin: a base scenario before the injection of CO2, and a repeat scenario corresponding to a P-wave velocity decline in the reservoir by 20 percent. For both scenarios, we simulate passive seismic experiments of body-wave noise recordings that may take several days or months to record in the field. The passive recordings are obtained by modelling global (direct wave, internal and surface multiples) transmission responses from band-limited subsurface noise sources, randomly triggered in space and time. The time-lapse reflection signal is obtained by taking the differences between the base and the repeat retrieved reflection data (virtual common-shot gathers). We found that the time-lapse signal is still recovered with ANSI even if the base and repeat retrieved reflection data are partially polluted with artifacts. This means that uneven illumination of the array does not

  6. Incubator-independent cell-culture perfusion platform for continuous long-term microelectrode array electrophysiology and time-lapse imaging

    PubMed Central

    Saalfrank, Dirk; Konduri, Anil Krishna; Latifi, Shahrzad; Habibey, Rouhollah; Golabchi, Asiyeh; Martiniuc, Aurel Vasile; Knoll, Alois; Ingebrandt, Sven; Blau, Axel

    2015-01-01

    Most in vitro electrophysiology studies extract information and draw conclusions from representative, temporally limited snapshot experiments. This approach bears the risk of missing decisive moments that may make a difference in our understanding of physiological events. This feasibility study presents a simple benchtop cell-culture perfusion system adapted to commercial microelectrode arrays (MEAs), multichannel electrophysiology equipment and common inverted microscopy stages for simultaneous and uninterrupted extracellular electrophysiology and time-lapse imaging at ambient CO2 levels. The concept relies on a transparent, replica-casted polydimethylsiloxane perfusion cap, gravity- or syringe-pump-driven perfusion and preconditioning of pH-buffered serum-free cell-culture medium to ambient CO2 levels at physiological temperatures. The low-cost microfluidic in vitro enabling platform, which allows us to image cultures immediately after cell plating, is easy to reproduce and is adaptable to the geometries of different cell-culture containers. It permits the continuous and simultaneous multimodal long-term acquisition or manipulation of optical and electrophysiological parameter sets, thereby considerably widening the range of experimental possibilities. Two exemplary proof-of-concept long-term MEA studies on hippocampal networks illustrate system performance. Continuous extracellular recordings over a period of up to 70 days revealed details on both sudden and gradual neural activity changes in maturing cell ensembles with large intra-day fluctuations. Correlated time-lapse imaging unveiled rather static macroscopic network architectures with previously unreported local morphological oscillations on the timescale of minutes. PMID:26543581

  7. Multiplexed time-lapse photomicrography of cultured cells.

    PubMed

    Heye, R R; Kiebler, E W; Arnzen, R J; Tolmach, L J

    1982-01-01

    A system of cinemicrography has been developed in which a single microscope and 16 mm camera are multiplexed to produce a time-lapse photographic record of many fields simultaneously. The field coordinates and focus are selected via a control console and entered into the memory of a dedicated microcomputer; they are then automatically recalled in sequence, thus permitting the photographing of additional fields in the interval between exposures of any given field. Sequential exposures of each field are isolated in separate sections of the film by means of a specially designed random-access camera that is also controlled by the microcomputer. The need to unscramble frames is thereby avoided, and the developed film can be directly analysed.

  8. Time-lapse electrical resistivity tomography: a powerful tool for landslide monitoring?

    NASA Astrophysics Data System (ADS)

    Perrone, A.

    2011-12-01

    The extreme rainfall events and the quick snowmelt occurrences play an important role in the triggering of the landslides. The occurrence of one of these factors can determine the variation of water content in the first layers of the subsoil and as a consequence a quick soil saturation inducing both an increase in pore-water pressures and the overloaded of the slopes progressively collapsing. The electrical resistivity, self-potential, electromagnetic induction and GPR methods can be considered as the most appropriate for assessing the presence of water in the underground. Such methods allow us to study the behavior of water content over much wider and deeper areas than those offered by traditional methods (thermo-gravimetric, tensiometric, TDR, etc) based on spot measures and concerning small volumes. In particular, the Electrical Resistivity Tomography (ERT), which has already proved to be a powerful tool both for the geometrical reconstruction of a landslide body (location of sliding surface, estimation of the thickness of the slide material) and the individuation of high water content areas, can be considered as an alternative tool to be employed for a qualitative and quantitative water content monitoring in the first layers of the subsoil. Indeed, time-lapse 2D ERT can be tested in order to gather information on the temporal and spatial patterns of water infiltration processes and water content variation. This work reports the preliminary results from a new prototype system planned to obtain time-lapse 2D ERTs, TDR and precipitation measurements in two landslide areas located in the Southern Apennine chain (Italy). The system was planned with the aim to estimate the variation of the resistivity parameter on a long period considering the water content variation, the rain water infiltration and the seasonal changes. The prototype system, linked to a pc used for storing data and managing the time interval acquisition, consists of: a resistivimeter connected to a

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

    NASA Astrophysics Data System (ADS)

    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.

  10. Early Lapses in a Cessation Attempt: Lapse Contexts, Cessation Success, and Predictors of Early Lapse

    PubMed Central

    2013-01-01

    Introduction: The great majority of smokers relapse when they make quit attempts. Therefore, understanding the process of relapse may guide the development of more effective smoking cessation or relapse prevention treatments. The goal of this research is to extend our understanding of the context of initial lapses that occur within 8 weeks of quitting by using more comprehensive assessments of context, a contemporary sample, and sophisticated analytic techniques. Methods: Participants from a randomized controlled smoking cessation trial completed baseline assessments of demographics and tobacco dependence, a daily smoking calendar to determine latency to lapse and relapse (7 consecutive days of smoking), and an assessment of initial lapse context (affect, location, activity, interpersonal, smoke exposure, and cigarette availability). Latent class analysis (LCA) was used to analyze the 6 early lapse (within the first 8 weeks; N = 551) context dimensions; logistic regression and Cox regression were used to relate context to cessation outcomes. Results: LCA revealed 5 distinct initial lapse context classes (talking, with friends, angry; social; alone; with spouse, angry; and with smoking spouse) that were differentially related to cessation outcome. The easy availability of cigarettes characterized almost 75% of lapses, but being with friends, drinking, and not being at home were associated with a lower likelihood of progression to relapse. Conclusions: Early lapsing is highly related to ultimate relapse, and lapsing in frequently experienced contexts seemed most strongly linked with progression to full relapse. PMID:23780705

  11. Time-lapse and slow-motion tracking of temperature changes: response time of a thermometer

    NASA Astrophysics Data System (ADS)

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

    2017-03-01

    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 in a simple yet rigorous way the response time of an alcohol thermometer and show its critical dependence on the properties of the surrounding environment giving insight into instrument characteristics, heat transfer and thermal equilibrium concepts.

  12. Time-lapse analysis of potential cellular responsiveness to Johrei, a Japanese healing technique

    PubMed Central

    Taft, Ryan; Moore, Dan; Yount, Garret

    2005-01-01

    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. PMID:15667653

  13. High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy

    NASA Astrophysics Data System (ADS)

    Guo, Baoshan; Lei, Cheng; Ito, Takuro; Yaxiaer, Yalikun; Kobayashi, Hirofumi; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

    2017-02-01

    The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, microalgal biofuel is expected to play a key role in reducing the detrimental effects of global warming since microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid contents and fail to characterize a diverse population of microalgal cells with single-cell resolution in a noninvasive and interference-free manner. Here we demonstrate high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy. In particular, we use Euglena gracilis - an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement) within lipid droplets. Our optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch phase-contrast microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase contents of every single cell at a high throughput of 10,000 cells/s. We characterize heterogeneous populations of E. gracilis cells under two different culture conditions to evaluate their lipid production efficiency. Our method holds promise as an effective analytical tool for microalgaebased biofuel production.

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

    NASA Technical Reports Server (NTRS)

    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.

  15. Extended Field Laser Confocal Microscopy (EFLCM): Combining automated Gigapixel image capture with in silico virtual microscopy

    PubMed Central

    Flaberg, Emilie; Sabelström, Per; Strandh, Christer; Szekely, Laszlo

    2008-01-01

    Background Confocal laser scanning microscopy has revolutionized cell biology. However, the technique has major limitations in speed and sensitivity due to the fact that a single laser beam scans the sample, allowing only a few microseconds signal collection for each pixel. This limitation has been overcome by the introduction of parallel beam illumination techniques in combination with cold CCD camera based image capture. Methods Using the combination of microlens enhanced Nipkow spinning disc confocal illumination together with fully automated image capture and large scale in silico image processing we have developed a system allowing the acquisition, presentation and analysis of maximum resolution confocal panorama images of several Gigapixel size. We call the method Extended Field Laser Confocal Microscopy (EFLCM). Results We show using the EFLCM technique that it is possible to create a continuous confocal multi-colour mosaic from thousands of individually captured images. EFLCM can digitize and analyze histological slides, sections of entire rodent organ and full size embryos. It can also record hundreds of thousands cultured cells at multiple wavelength in single event or time-lapse fashion on fixed slides, in live cell imaging chambers or microtiter plates. Conclusion The observer independent image capture of EFLCM allows quantitative measurements of fluorescence intensities and morphological parameters on a large number of cells. EFLCM therefore bridges the gap between the mainly illustrative fluorescence microscopy and purely quantitative flow cytometry. EFLCM can also be used as high content analysis (HCA) instrument for automated screening processes. PMID:18627634

  16. Extended Field Laser Confocal Microscopy (EFLCM): combining automated Gigapixel image capture with in silico virtual microscopy.

    PubMed

    Flaberg, Emilie; Sabelström, Per; Strandh, Christer; Szekely, Laszlo

    2008-07-16

    Confocal laser scanning microscopy has revolutionized cell biology. However, the technique has major limitations in speed and sensitivity due to the fact that a single laser beam scans the sample, allowing only a few microseconds signal collection for each pixel. This limitation has been overcome by the introduction of parallel beam illumination techniques in combination with cold CCD camera based image capture. Using the combination of microlens enhanced Nipkow spinning disc confocal illumination together with fully automated image capture and large scale in silico image processing we have developed a system allowing the acquisition, presentation and analysis of maximum resolution confocal panorama images of several Gigapixel size. We call the method Extended Field Laser Confocal Microscopy (EFLCM). We show using the EFLCM technique that it is possible to create a continuous confocal multi-colour mosaic from thousands of individually captured images. EFLCM can digitize and analyze histological slides, sections of entire rodent organ and full size embryos. It can also record hundreds of thousands cultured cells at multiple wavelength in single event or time-lapse fashion on fixed slides, in live cell imaging chambers or microtiter plates. The observer independent image capture of EFLCM allows quantitative measurements of fluorescence intensities and morphological parameters on a large number of cells. EFLCM therefore bridges the gap between the mainly illustrative fluorescence microscopy and purely quantitative flow cytometry. EFLCM can also be used as high content analysis (HCA) instrument for automated screening processes.

  17. Multimodal computational microscopy based on transport of intensity equation

    NASA Astrophysics Data System (ADS)

    Li, Jiaji; Chen, Qian; Sun, Jiasong; Zhang, Jialin; Zuo, Chao

    2016-12-01

    Transport of intensity equation (TIE) is a powerful tool for phase retrieval and quantitative phase imaging, which requires intensity measurements only at axially closely spaced planes without a separate reference beam. It does not require coherent illumination and works well on conventional bright-field microscopes. The quantitative phase reconstructed by TIE gives valuable information that has been encoded in the complex wave field by passage through a sample of interest. Such information may provide tremendous flexibility to emulate various microscopy modalities computationally without requiring specialized hardware components. We develop a requisite theory to describe such a hybrid computational multimodal imaging system, which yields quantitative phase, Zernike phase contrast, differential interference contrast, and light field moment imaging, simultaneously. It makes the various observations for biomedical samples easy. Then we give the experimental demonstration of these ideas by time-lapse imaging of live HeLa cell mitosis. Experimental results verify that a tunable lens-based TIE system, combined with the appropriate postprocessing algorithm, can achieve a variety of promising imaging modalities in parallel with the quantitative phase images for the dynamic study of cellular processes.

  18. Very-high-resolution time-lapse photography for plant and ecosystems research1

    PubMed Central

    Nichols, Mary H.; Steven, Janet C.; Sargent, Randy; Dille, Paul; Schapiro, Joshua

    2013-01-01

    • Premise of the study: Traditional photography is a compromise between image detail and area covered. We report a new method for creating time-lapse sequences of very-high-resolution photographs to produce zoomable images that facilitate observation across a range of spatial and temporal scales. • Methods and Results: A robotic camera mount and software were used to capture images of the growth and movement in Brassica rapa every 15 s in the laboratory. The resultant time-lapse sequence (http://timemachine.gigapan.org/wiki/Plant_Growth) captures growth detail such as circumnutation. A modified, solar-powered system was deployed at a remote field site in southern Arizona. Images were collected every 2 h over a 3-mo period to capture the response of vegetation to monsoon season rainfall (http://timemachine.gigapan.org/wiki/Arizona_Grasslands). • Conclusions: A technique for observing time sequences of both individual plant and ecosystem response at a range of spatial scales is available for use in the laboratory and in the field. PMID:25202588

  19. Selective neuronal lapses precede human cognitive lapses following sleep deprivation.

    PubMed

    Nir, Yuval; Andrillon, Thomas; Marmelshtein, Amit; Suthana, Nanthia; Cirelli, Chiara; Tononi, Giulio; Fried, Itzhak

    2017-12-01

    Sleep deprivation is a major source of morbidity with widespread health effects, including increased risk of hypertension, diabetes, obesity, heart attack, and stroke. Moreover, sleep deprivation brings about vehicle accidents and medical errors and is therefore an urgent topic of investigation. During sleep deprivation, homeostatic and circadian processes interact to build up sleep pressure, which results in slow behavioral performance (cognitive lapses) typically attributed to attentional thalamic and frontoparietal circuits, but the underlying mechanisms remain unclear. Recently, through study of electroencephalograms (EEGs) in humans and local field potentials (LFPs) in nonhuman primates and rodents it was found that, during sleep deprivation, regional 'sleep-like' slow and theta (slow/theta) waves co-occur with impaired behavioral performance during wakefulness. Here we used intracranial electrodes to record single-neuron activities and LFPs in human neurosurgical patients performing a face/nonface categorization psychomotor vigilance task (PVT) over multiple experimental sessions, including a session after full-night sleep deprivation. We find that, just before cognitive lapses, the selective spiking responses of individual neurons in the medial temporal lobe (MTL) are attenuated, delayed, and lengthened. These 'neuronal lapses' are evident on a trial-by-trial basis when comparing the slowest behavioral PVT reaction times to the fastest. Furthermore, during cognitive lapses, LFPs exhibit a relative local increase in slow/theta activity that is correlated with degraded single-neuron responses and with baseline theta activity. Our results show that cognitive lapses involve local state-dependent changes in neuronal activity already present in the MTL.

  20. Selected time-lapse movies of the east rift zone eruption of KĪlauea Volcano, 2004–2008

    USGS Publications Warehouse

    Orr, Tim R.

    2011-01-01

    Since 2004, the U.S. Geological Survey's Hawaiian Volcano Observatory has used mass-market digital time-lapse cameras and network-enabled Webcams for visual monitoring and research. The 26 time-lapse movies in this report were selected from the vast collection of images acquired by these camera systems during 2004–2008. Chosen for their content and broad aesthetic appeal, these image sequences document a variety of flow-field and vent processes from Kīlauea's east rift zone eruption, which began in 1983 and is still (as of 2011) ongoing.

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

    PubMed

    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.

  2. The sexual phase of the diatom Pseudo-nitzschia multistriata: cytological and time-lapse cinematography characterization.

    PubMed

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

    PubMed

    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.

  4. Reconstructed imaging of acoustic cloak using time-lapse reversal method

    NASA Astrophysics Data System (ADS)

    Zhou, Chen; Cheng, Ying; Xu, Jian-yi; Li, Bo; Liu, Xiao-jun

    2014-08-01

    We proposed and investigated a solution to the inverse acoustic cloak problem, an anti-stealth technology to make cloaks visible, using the time-lapse reversal (TLR) method. The TLR method reconstructs the image of an unknown acoustic cloak by utilizing scattered acoustic waves. Compared to previous anti-stealth methods, the TLR method can determine not only the existence of a cloak but also its exact geometric information like definite shape, size, and position. Here, we present the process for TLR reconstruction based on time reversal invariance. This technology may have potential applications in detecting various types of cloaks with different geometric parameters.

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

    PubMed

    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.

  6. Time-Lapse Motion Picture Technique Applied to the Study of Geological Processes.

    PubMed

    Miller, R D; Crandell, D R

    1959-09-25

    Light-weight, battery-operated timers were built and coupled to 16-mm motion-picture cameras having apertures controlled by photoelectric cells. The cameras were placed adjacent to Emmons Glacier on Mount Rainier. The film obtained confirms the view that exterior time-lapse photography can be applied to the study of slow-acting geologic processes.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Supriyadi,, E-mail: supriyadi@mail.unnes.ac.id; Khumaedi; Yusuf, M.

    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 weremore » 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.« less

  8. In vivo time-lapse imaging of cell proliferation and differentiation in the optic tectum of Xenopus laevis tadpoles

    PubMed Central

    Bestman, Jennifer E.; Lee-Osbourne, Jane; Cline, Hollis T.

    2012-01-01

    We analyzed the function of neural progenitors in the developing CNS of Xenopus laevis tadpoles using in vivo time-lapse confocal microscopy to collect images through the tectum at intervals of 2 to 24 hours over 3 days. Neural progenitor cells were labeled with fluorescent protein reporters based on expression of endogenous Sox2 transcription factor. With this construct, we identified Sox2-expressing cells as radial glia and as a component of the progenitor pool of cells in the developing tectum that gives rise to neurons and other radial glia. Lineage analysis of individual radial glia and their progeny demonstrated that less than 10% of radial glia undergo symmetric divisions resulting in two radial glia, while the majority of radial glia divide asymmetrically to generate neurons and radial glia. Time-lapse imaging revealed the direct differentiation of radial glia into neurons. Although radial glia may guide axons as they navigate to superficial tectum, we find no evidence that radial glia function as a scaffold for neuronal migration at early stages of tectal development. Over three days, the number of labeled cells increased 20%, as the fraction of radial glia dropped and the proportion of neuronal progeny increased to approximately 60% of the labeled cells. Tadpoles provided with short-term visual enhancement generated significantly more neurons, with a corresponding decrease in cell proliferation. Together these results demonstrate that radial glial cells are neural progenitors in the developing optic tectum and reveal that visual experience increases the proportion of neurons generated in an intact animal. PMID:22113462

  9. High-speed transport-of-intensity phase microscopy with an electrically tunable lens.

    PubMed

    Zuo, Chao; Chen, Qian; Qu, Weijuan; Asundi, Anand

    2013-10-07

    We present a high-speed transport-of-intensity equation (TIE) quantitative phase microscopy technique, named TL-TIE, by combining an electrically tunable lens with a conventional transmission microscope. This permits the specimen at different focus position to be imaged in rapid succession, with constant magnification and no physically moving parts. The simplified image stack collection significantly reduces the acquisition time, allows for the diffraction-limited through-focus intensity stack collection at 15 frames per second, making dynamic TIE phase imaging possible. The technique is demonstrated by profiling of microlens array using optimal frequency selection scheme, and time-lapse imaging of live breast cancer cells by inversion the defocused phase optical transfer function to correct the phase blurring in traditional TIE. Experimental results illustrate its outstanding capability of the technique for quantitative phase imaging, through a simple, non-interferometric, high-speed, high-resolution, and unwrapping-free approach with prosperous applications in micro-optics, life sciences and bio-photonics.

  10. In Vivo Time-Lapse Imaging in the Zebrafish Lateral Line: A Flexible, Open-Ended Research Project for an Undergraduate Neurobiology Laboratory Course.

    PubMed

    Marra, Molly H; Tobias, Zachary J C; Cohen, Hannah R; Glover, Greta; Weissman, Tamily A

    2015-01-01

    The lateral line sensory system in fish detects movements in the water and allows fish to respond to predators, prey, and other stimuli. As the lateral line forms in the first two days of zebrafish development, axons extend caudally along the lateral surface of the fish, eventually forming synapses with hair cells of neuromasts. Growing lateral line axons are located superficially under the skin and can be labeled in living zebrafish using fluorescent protein expression. This system provides a relatively straightforward approach for in vivo time-lapse imaging of neuronal development in an undergraduate setting. Here we describe an upper-level neurobiology laboratory module in which students investigate aspects of axonal development in the zebrafish lateral line system. Students learn to handle and image living fish, collect time-lapse videos of moving mitochondria, and quantitatively measure mitochondrial dynamics by generating and analyzing kymographs of their movements. Energy demands may differ between axons with extending growth cones versus axons that have already reached their targets and are forming synapses. Since relatively little is known about this process in developing lateral line axons, students generate and test their own hypotheses regarding how mitochondrial dynamics may differ at two different time points in axonal development. Students also learn to incorporate into their analysis a powerful yet accessible quantitative tool, the kymograph, which is used to graph movement over time. After students measure and quantify dynamics in living fish at 1 and 2 days post fertilization, this module extends into independent projects, in which students can expand their studies in a number of different, inquiry-driven directions. The project can also be pared down for courses that wish to focus solely on the quantitative analysis (without fish handling), or vice versa. This research module provides a useful approach for the design of open-ended laboratory

  11. Integral refractive index imaging of flowing cell nuclei using quantitative phase microscopy combined with fluorescence microscopy.

    PubMed

    Dardikman, Gili; Nygate, Yoav N; Barnea, Itay; Turko, Nir A; Singh, Gyanendra; Javidi, Barham; Shaked, Natan T

    2018-03-01

    We suggest a new multimodal imaging technique for quantitatively measuring the integral (thickness-average) refractive index of the nuclei of live biological cells in suspension. For this aim, we combined quantitative phase microscopy with simultaneous 2-D fluorescence microscopy. We used 2-D fluorescence microscopy to localize the nucleus inside the quantitative phase map of the cell, as well as for measuring the nucleus radii. As verified offline by both 3-D confocal fluorescence microscopy and 2-D fluorescence microscopy while rotating the cells during flow, the nucleus of cells in suspension that are not during division can be assumed to be an ellipsoid. The entire shape of a cell in suspension can be assumed to be a sphere. Then, the cell and nucleus 3-D shapes can be evaluated based on their in-plain radii available from the 2-D phase and fluorescent measurements, respectively. Finally, the nucleus integral refractive index profile is calculated. We demonstrate the new technique on cancer cells, obtaining nucleus refractive index values that are lower than those of the cytoplasm, coinciding with recent findings. We believe that the proposed technique has the potential to be used for flow cytometry, where full 3-D refractive index tomography is too slow to be implemented during flow.

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

    NASA Astrophysics Data System (ADS)

    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

  13. Fast history matching of time-lapse seismic and production data for high resolution models

    NASA Astrophysics Data System (ADS)

    Jimenez Arismendi, Eduardo Antonio

    Integrated reservoir modeling has become an important part of day-to-day decision analysis in oil and gas management practices. A very attractive and promising technology is the use of time-lapse or 4D seismic as an essential component in subsurface modeling. Today, 4D seismic is enabling oil companies to optimize production and increase recovery through monitoring fluid movements throughout the reservoir. 4D seismic advances are also being driven by an increased need by the petroleum engineering community to become more quantitative and accurate in our ability to monitor reservoir processes. Qualitative interpretations of time-lapse anomalies are being replaced by quantitative inversions of 4D seismic data to produce accurate maps of fluid saturations, pore pressure, temperature, among others. Within all steps involved in this subsurface modeling process, the most demanding one is integrating the geologic model with dynamic field data, including 4Dseismic when available. The validation of the geologic model with observed dynamic data is accomplished through a "history matching" (HM) process typically carried out with well-based measurements. Due to low resolution of production data, the validation process is severely limited in its reservoir areal coverage, compromising the quality of the model and any subsequent predictive exercise. This research will aim to provide a novel history matching approach that can use information from high-resolution seismic data to supplement the areally sparse production data. The proposed approach will utilize streamline-derived sensitivities as means of relating the forward model performance with the prior geologic model. The essential ideas underlying this approach are similar to those used for high-frequency approximations in seismic wave propagation. In both cases, this leads to solutions that are defined along "streamlines" (fluid flow), or "rays" (seismic wave propagation). Synthetic and field data examples will be used

  14. Quantitative DIC microscopy using an off-axis self-interference approach.

    PubMed

    Fu, Dan; Oh, Seungeun; Choi, Wonshik; Yamauchi, Toyohiko; Dorn, August; Yaqoob, Zahid; Dasari, Ramachandra R; Feld, Michael S

    2010-07-15

    Traditional Normarski differential interference contrast (DIC) microscopy is a very powerful method for imaging nonstained biological samples. However, one of its major limitations is the nonquantitative nature of the imaging. To overcome this problem, we developed a quantitative DIC microscopy method based on off-axis sample self-interference. The digital holography algorithm is applied to obtain quantitative phase gradients in orthogonal directions, which leads to a quantitative phase image through a spiral integration of the phase gradients. This method is practically simple to implement on any standard microscope without stringent requirements on polarization optics. Optical sectioning can be obtained through enlarged illumination NA.

  15. Composite time-lapse computed tomography and micro finite element simulations: A new imaging approach for characterizing cement flows and mechanical benefits of vertebroplasty.

    PubMed

    Stadelmann, Vincent A; Zderic, Ivan; Baur, Annick; Unholz, Cynthia; Eberli, Ursula; Gueorguiev, Boyko

    2016-02-01

    Vertebroplasty has been shown to reinforce weak vertebral bodies and reduce fracture risks, yet cement leakage is a major problem that can cause severe complications. Since cement flow is nearly impossible to control during surgery, small volumes of cement are injected, but then mechanical benefits might be limited. A better understanding of cement flows within bone structure is required to further optimize vertebroplasty and bone augmentation in general. We developed a novel imaging method, composite time-lapse CT, to characterize cement flow during injection. In brief, composite-resolution time-lapse CT exploits the qualities of microCT and clinical CT. The method consists in overlaying low-resolution time-lapse CT scans acquired during injection onto pre-operative high-resolution microCT scans, generating composite-resolution time-lapse CT series of cement flow within bone. In this in vitro study, composite-resolution time-lapse CT was applied to eight intact and five artificially fractured cadaveric vertebrae during vertebroplasty. The time-lapse scans were acquired at one-milliliter cement injection steps until a total of 10 ml cement was injected. The composite-resolution series were then converted into micro finite element models to compute strains distribution under virtual axial loading. Relocation of strain energy density within bone structure was observed throughout the progression of the procedure. Interestingly, the normalized effect of cement injection on the overall stiffness of the vertebrae was similar between intact and fractured specimens, although at different orders of magnitude. In conclusion, composite time-lapse CT can picture cement flows during bone augmentation. The composite images can also be easily converted into finite element models to compute virtual strain distributions under loading at every step of an injection, providing deeper understanding on the biomechanics of vertebroplasty. Copyright © 2015 IPEM. Published by Elsevier Ltd

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

    NASA Astrophysics Data System (ADS)

    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. Quantitative evaluation of morphological changes in activated platelets in vitro using digital holographic microscopy.

    PubMed

    Kitamura, Yutaka; Isobe, Kazushige; Kawabata, Hideo; Tsujino, Tetsuhiro; Watanabe, Taisuke; Nakamura, Masayuki; Toyoda, Toshihisa; Okudera, Hajime; Okuda, Kazuhiro; Nakata, Koh; Kawase, Tomoyuki

    2018-06-18

    Platelet activation and aggregation have been conventionally evaluated using an aggregometer. However, this method is suitable for short-term but not long-term quantitative evaluation of platelet aggregation, morphological changes, and/or adhesion to specific materials. The recently developed digital holographic microscopy (DHM) has enabled the quantitative evaluation of cell size and morphology without labeling or destruction. Thus, we aim to validate its applicability in quantitatively evaluating changes in cell morphology, especially in the aggregation and spreading of activated platelets, thus modifying typical image analysis procedures to suit aggregated platelets. Freshly prepared platelet-rich plasma was washed with phosphate-buffered saline and treated with 0.1% CaCl 2 . Platelets were then fixed and subjected to DHM, scanning electron microscopy (SEM), atomic force microscopy, optical microscopy, and flow cytometry (FCM). Tightly aggregated platelets were identified as single cells. Data obtained from time-course experiments were plotted two-dimensionally according to the average optical thickness versus attachment area and divided into four regions. The majority of the control platelets, which supposedly contained small and round platelets, were distributed in the lower left region. As activation time increased, however, this population dispersed toward the upper right region. The distribution shift demonstrated by DHM was essentially consistent with data obtained from SEM and FCM. Therefore, DHM was validated as a promising device for testing platelet function given that it allows for the quantitative evaluation of activation-dependent morphological changes in platelets. DHM technology will be applicable to the quality assurance of platelet concentrates, as well as diagnosis and drug discovery related to platelet functions. Copyright © 2018 Elsevier Ltd. All rights reserved.

  18. Monitoring channel head erosion processes in response to an artificially induced abrupt base level change using time-lapse photography

    NASA Astrophysics Data System (ADS)

    Nichols, M. H.; Nearing, M.; Hernandez, M.; Polyakov, V. O.

    2016-07-01

    Gullies that terminate at a vertical-wall are ubiquitous throughout arid and semiarid regions. Multi-year assessments of gully evolution and headcut advance are typically accomplished using traditional ground surveys and aerial photographs, with much recent research focused on integrating data collected at very high spatial resolutions using new techniques such as aerial surveys with blimps or kites and ground surveys with LiDar scanners. However, knowledge of specific processes that drive headcut advance is limited due to inadequate observation and documentation of flash floods and subsequent erosion that can occur at temporal resolutions not captured through repeat surveys. This paper presents a method for using very-high temporal resolution ground-based time-lapse photography to capture short-duration flash floods and gully head evolution in response. In 2004, a base level controlling concrete weir was removed from the outlet of a 1.29 ha semiarid headwater drainage on the Walnut Gulch Experimental Watershed in southeastern Arizona, USA. During the ten year period from 2004 to 2014 the headcut migrated upchannel a total of 14.5 m reducing the contributing area at the headwall by 9.5%. Beginning in July 2012, time-lapse photography was employed to observe event scale channel evolution dynamics. The most frequent erosion processes observed during three seasons of time-lapse photography were plunge pool erosion and mass wasting through sidewall or channel headwall slumping that occurred during summer months. Geomorphic change during the ten year period was dominated by a single piping event in August 2014 that advanced the channel head 7.4 m (51% of the overall advance) and removed 11.3 m3 of sediment. High temporal resolution time-lapse photography was critical for identifying subsurface erosion processes, in the absence of time-lapse images piping would not have been identified as an erosion mechanism responsible for advancing the gully headwall at this site.

  19. Surface plasmon resonance microscopy: achieving a quantitative optical response

    PubMed Central

    Peterson, Alexander W.; Halter, Michael; Plant, Anne L.; Elliott, John T.

    2016-01-01

    Surface plasmon resonance (SPR) imaging allows real-time label-free imaging based on index of refraction, and changes in index of refraction at an interface. Optical parameter analysis is achieved by application of the Fresnel model to SPR data typically taken by an instrument in a prism based configuration. We carry out SPR imaging on a microscope by launching light into a sample, and collecting reflected light through a high numerical aperture microscope objective. The SPR microscope enables spatial resolution that approaches the diffraction limit, and has a dynamic range that allows detection of subnanometer to submicrometer changes in thickness of biological material at a surface. However, unambiguous quantitative interpretation of SPR changes using the microscope system could not be achieved using the Fresnel model because of polarization dependent attenuation and optical aberration that occurs in the high numerical aperture objective. To overcome this problem, we demonstrate a model to correct for polarization diattenuation and optical aberrations in the SPR data, and develop a procedure to calibrate reflectivity to index of refraction values. The calibration and correction strategy for quantitative analysis was validated by comparing the known indices of refraction of bulk materials with corrected SPR data interpreted with the Fresnel model. Subsequently, we applied our SPR microscopy method to evaluate the index of refraction for a series of polymer microspheres in aqueous media and validated the quality of the measurement with quantitative phase microscopy. PMID:27782542

  20. Time-lapse Sequence of Jupiter's South Pole

    NASA Image and Video Library

    2018-02-22

    This series of images captures cloud patterns near Jupiter's south pole, looking up towards the planet's equator. NASA's Juno spacecraft took the color-enhanced time-lapse sequence of images during its eleventh close flyby of the gas giant planet on Feb. 7 between 7:21 a.m. and 8:01 a.m. PST (10:21 a.m. and 11:01 a.m. EST). At the time, the spacecraft was between 85,292 to 124,856 miles (137,264 to 200,937 kilometers) from the tops of the clouds of the planet with the images centered on latitudes from 84.1 to 75.5 degrees south. At first glance, the series might appear to be the same image repeated. But closer inspection reveals slight changes, which are most easily noticed by comparing the far left image with the far right image. Directly, the images show Jupiter. But, through slight variations in the images, they indirectly capture the motion of the Juno spacecraft itself, once again swinging around a giant planet hundreds of millions of miles from Earth. https://photojournal.jpl.nasa.gov/catalog/PIA21979

  1. MATtrack: A MATLAB-Based Quantitative Image Analysis Platform for Investigating Real-Time Photo-Converted Fluorescent Signals in Live Cells

    PubMed Central

    Courtney, Jane; Woods, Elena; Scholz, Dimitri; Hall, William W.; Gautier, Virginie W.

    2015-01-01

    We introduce here MATtrack, an open source MATLAB-based computational platform developed to process multi-Tiff files produced by a photo-conversion time lapse protocol for live cell fluorescent microscopy. MATtrack automatically performs a series of steps required for image processing, including extraction and import of numerical values from Multi-Tiff files, red/green image classification using gating parameters, noise filtering, background extraction, contrast stretching and temporal smoothing. MATtrack also integrates a series of algorithms for quantitative image analysis enabling the construction of mean and standard deviation images, clustering and classification of subcellular regions and injection point approximation. In addition, MATtrack features a simple user interface, which enables monitoring of Fluorescent Signal Intensity in multiple Regions of Interest, over time. The latter encapsulates a region growing method to automatically delineate the contours of Regions of Interest selected by the user, and performs background and regional Average Fluorescence Tracking, and automatic plotting. Finally, MATtrack computes convenient visualization and exploration tools including a migration map, which provides an overview of the protein intracellular trajectories and accumulation areas. In conclusion, MATtrack is an open source MATLAB-based software package tailored to facilitate the analysis and visualization of large data files derived from real-time live cell fluorescent microscopy using photoconvertible proteins. It is flexible, user friendly, compatible with Windows, Mac, and Linux, and a wide range of data acquisition software. MATtrack is freely available for download at eleceng.dit.ie/courtney/MATtrack.zip. PMID:26485569

  2. MATtrack: A MATLAB-Based Quantitative Image Analysis Platform for Investigating Real-Time Photo-Converted Fluorescent Signals in Live Cells.

    PubMed

    Courtney, Jane; Woods, Elena; Scholz, Dimitri; Hall, William W; Gautier, Virginie W

    2015-01-01

    We introduce here MATtrack, an open source MATLAB-based computational platform developed to process multi-Tiff files produced by a photo-conversion time lapse protocol for live cell fluorescent microscopy. MATtrack automatically performs a series of steps required for image processing, including extraction and import of numerical values from Multi-Tiff files, red/green image classification using gating parameters, noise filtering, background extraction, contrast stretching and temporal smoothing. MATtrack also integrates a series of algorithms for quantitative image analysis enabling the construction of mean and standard deviation images, clustering and classification of subcellular regions and injection point approximation. In addition, MATtrack features a simple user interface, which enables monitoring of Fluorescent Signal Intensity in multiple Regions of Interest, over time. The latter encapsulates a region growing method to automatically delineate the contours of Regions of Interest selected by the user, and performs background and regional Average Fluorescence Tracking, and automatic plotting. Finally, MATtrack computes convenient visualization and exploration tools including a migration map, which provides an overview of the protein intracellular trajectories and accumulation areas. In conclusion, MATtrack is an open source MATLAB-based software package tailored to facilitate the analysis and visualization of large data files derived from real-time live cell fluorescent microscopy using photoconvertible proteins. It is flexible, user friendly, compatible with Windows, Mac, and Linux, and a wide range of data acquisition software. MATtrack is freely available for download at eleceng.dit.ie/courtney/MATtrack.zip.

  3. Total internal reflection holographic microscopy (TIRHM) for quantitative phase characterization of cell-substrate adhesion

    NASA Astrophysics Data System (ADS)

    Ash, William Mason, III

    Total Internal Reflection Holographic Microscopy (TIRHM) combines near-field microscopy with digital holography to produce a new form of near-field phase microscopy. Using a prism in TIR as a near-field imager, the presence of microscopic organisms, cell-substrate interfaces, and adhesions, causes relative refractive index (RRI) and frustrated TIR (f-TIR) to modulate the object beam's evanescent wave phase front. Quantitative phase images of test specimens such as Amoeba proteus, Dictyostelium Discoideum and cells such as SKOV-3 ovarian cancer and 3T3 fibroblasts are produced without the need to introduce stains or fluorophores. The angular spectrum method of digital holography to compensate for tilt anamorphism due to the inclined TIR plane is also discussed. The results of this work conclusively demonstrate, for the first time, the integration of near-field microscopy with digital holography. The cellular images presented show a correlation between the physical extent of the Amoeba proteus plasma membrane and the adhesions that are quantitatively profiled by phase cross-sectioning of the holographic images obtained by digital holography. With its ability to quantitatively characterise cellular adhesion and motility, it is anticipated that TIRHM can be a tool for characterizing and combating cancer metastasis, as well as improving our understanding of morphogenesis and embryogenesis itself.

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

    PubMed

    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.

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

    PubMed Central

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

    2013-01-01

    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. PMID:23676900

  6. Fracture mechanics by three-dimensional crack-tip synchrotron X-ray microscopy

    PubMed Central

    Withers, P. J.

    2015-01-01

    To better understand the relationship between the nucleation and growth of defects and the local stresses and phase changes that cause them, we need both imaging and stress mapping. Here, we explore how this can be achieved by bringing together synchrotron X-ray diffraction and tomographic imaging. Conventionally, these are undertaken on separate synchrotron beamlines; however, instruments capable of both imaging and diffraction are beginning to emerge, such as ID15 at the European Synchrotron Radiation Facility and JEEP at the Diamond Light Source. This review explores the concept of three-dimensional crack-tip X-ray microscopy, bringing them together to probe the crack-tip behaviour under realistic environmental and loading conditions and to extract quantitative fracture mechanics information about the local crack-tip environment. X-ray diffraction provides information about the crack-tip stress field, phase transformations, plastic zone and crack-face tractions and forces. Time-lapse CT, besides providing information about the three-dimensional nature of the crack and its local growth rate, can also provide information as to the activation of extrinsic toughening mechanisms such as crack deflection, crack-tip zone shielding, crack bridging and crack closure. It is shown how crack-tip microscopy allows a quantitative measure of the crack-tip driving force via the stress intensity factor or the crack-tip opening displacement. Finally, further opportunities for synchrotron X-ray microscopy are explored. PMID:25624521

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

    PubMed

    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).

  8. Subsurface lateral preferential flow network revealed by time-lapse ground-penetrating radar in a hillslope

    NASA Astrophysics Data System (ADS)

    Guo, Li; Chen, Jin; Lin, Henry

    2014-12-01

    Subsurface lateral preferential flow (LPF) has been observed to contribute substantially to hillslope and catchment runoff. However, the complex nature of LPF and the lack of an appropriate investigation method have hindered direct LPF observation in the field. Thus, the initiation, persistence, and dynamics of LPF networks remain poorly understood. This study explored the application of time-lapse ground-penetrating radar (GPR) together with an artificial infiltration to shed light on the nature of LPF and its dynamics in a hillslope. Based on our enhanced field experimental setup and carefully refined GPR data postprocessing algorithms, we developed a new protocol to reconstruct LPF networks with centimeter resolution. This is the first time that a detailed LPF network and its dynamics have been revealed noninvasively along a hillslope. Real-time soil water monitoring and field soil investigation confirmed the locations of LPF mapped by time-lapse GPR surveys. Our results indicated the following: (1) Increased spatial variations of radar signals after infiltration suggested heterogeneous soil water changes within the studied soil, which reflected the generation and dynamics of LPF; (2) Two types of LPF networks were identified, the network at the location of soil permeability contrasts and that formed via a series of connected preferential flow paths; and (3) The formation and distribution of LPF networks were influenced by antecedent soil water condition. Overall, this study demonstrates clearly that carefully designed time-lapse GPR surveys with enhanced data postprocessing offer a practical and nondestructive way of mapping LPF networks in the field, thereby providing a potentially significant enhancement in our ability to study complex subsurface flow processes across the landscape.

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

    NASA Astrophysics Data System (ADS)

    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

  10. Arctic sea-ice variations from time-lapse passive microwave imagery

    USGS Publications Warehouse

    Campbell, W.J.; Ramseier, R.O.; Zwally, H.J.; Gloersen, P.

    1980-01-01

    This paper presents: (1) a short historical review of the passive microwave research on sea ice which established the observational and theoretical base permitting the interpretation of the first passive microwave images of Earth obtained by the Nimbus-5 ESMR; (2) the construction of a time-lapse motion picture film of a 16-month set of serial ESMR images to aid in the formidable data analysis task; and (3) a few of the most significant findings resulting from an early analysis of these data, using selected ESMR images to illustrate these findings. ?? 1980 D. Reidel Publishing Co.

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

    EPA Science Inventory

    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...

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

    USGS Publications Warehouse

    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.

  13. Lapse of time effects on tax evasion in an agent-based econophysics model

    NASA Astrophysics Data System (ADS)

    Seibold, Götz; Pickhardt, Michael

    2013-05-01

    We investigate an inhomogeneous Ising model in the context of tax evasion dynamics where different types of agents are parameterized via local temperatures and magnetic fields. In particular, we analyze the impact of lapse of time effects (i.e. backauditing) and endogenously determined penalty rates on tax compliance. Both features contribute to a microfoundation of agent-based econophysics models of tax evasion.

  14. 3D fluorescence anisotropy imaging using selective plane illumination microscopy.

    PubMed

    Hedde, Per Niklas; Ranjit, Suman; Gratton, Enrico

    2015-08-24

    Fluorescence anisotropy imaging is a popular method to visualize changes in organization and conformation of biomolecules within cells and tissues. In such an experiment, depolarization effects resulting from differences in orientation, proximity and rotational mobility of fluorescently labeled molecules are probed with high spatial resolution. Fluorescence anisotropy is typically imaged using laser scanning and epifluorescence-based approaches. Unfortunately, those techniques are limited in either axial resolution, image acquisition speed, or by photobleaching. In the last decade, however, selective plane illumination microscopy has emerged as the preferred choice for three-dimensional time lapse imaging combining axial sectioning capability with fast, camera-based image acquisition, and minimal light exposure. We demonstrate how selective plane illumination microscopy can be utilized for three-dimensional fluorescence anisotropy imaging of live cells. We further examined the formation of focal adhesions by three-dimensional time lapse anisotropy imaging of CHO-K1 cells expressing an EGFP-paxillin fusion protein.

  15. Quantitative Imaging of Single Unstained Magnetotactic Bacteria by Coherent X-ray Diffraction Microscopy.

    PubMed

    Fan, Jiadong; Sun, Zhibin; Zhang, Jian; Huang, Qingjie; Yao, Shengkun; Zong, Yunbing; Kohmura, Yoshiki; Ishikawa, Tetsuya; Liu, Hong; Jiang, Huaidong

    2015-06-16

    Novel coherent diffraction microscopy provides a powerful lensless imaging method to obtain a better understanding of the microorganism at the nanoscale. Here we demonstrated quantitative imaging of intact unstained magnetotactic bacteria using coherent X-ray diffraction microscopy combined with an iterative phase retrieval algorithm. Although the signal-to-noise ratio of the X-ray diffraction pattern from single magnetotactic bacterium is weak due to low-scattering ability of biomaterials, an 18.6 nm half-period resolution of reconstructed image was achieved by using a hybrid input-output phase retrieval algorithm. On the basis of the quantitative reconstructed images, the morphology and some intracellular structures, such as nucleoid, polyβ-hydroxybutyrate granules, and magnetosomes, were identified, which were also confirmed by scanning electron microscopy and energy dispersive spectroscopy. With the benefit from the quantifiability of coherent diffraction imaging, for the first time to our knowledge, an average density of magnetotactic bacteria was calculated to be ∼1.19 g/cm(3). This technique has a wide range of applications, especially in quantitative imaging of low-scattering biomaterials and multicomponent materials at nanoscale resolution. Combined with the cryogenic technique or X-ray free electron lasers, the method could image cells in a hydrated condition, which helps to maintain their natural structure.

  16. Time-lapse processing of 2D seismic profiles with testing of static correction methods at the CO2 injection site Ketzin (Germany)

    NASA Astrophysics Data System (ADS)

    Bergmann, Peter; Yang, Can; Lüth, Stefan; Juhlin, Christopher; Cosma, Calin

    2011-09-01

    The Ketzin project provides an experimental pilot test site for the geological storage of CO2. Seismic monitoring of the Ketzin site comprises 2D and 3D time-lapse experiments with baseline experiments in 2005. The first repeat 2D survey was acquired in 2009 after 22 kt of CO2 had been injected into the Stuttgart Formation at approximately 630 m depth. Main objectives of the 2D seismic surveys were the imaging of geological structures, detection of injected CO2, and comparison with the 3D surveys. Time-lapse processing highlighted the importance of detailed static corrections to account for travel time delays, which are attributed to different near-surface velocities during the survey periods. Compensation for these delays has been performed using both pre-stack static corrections and post-stack static corrections. The pre-stack method decomposes the travel time delays of baseline and repeat datasets in a surface consistent manner, while the latter cross-aligns baseline and repeat stacked sections along a reference horizon. Application of the static corrections improves the S/N ratio of the time-lapse sections significantly. Based on our results, it is recommended to apply a combination of both corrections when time-lapse processing faces considerable near-surface velocity changes. Processing of the datasets demonstrates that the decomposed solution of the pre-stack static corrections can be used for interpretation of changes in near-surface velocities. In particular, the long-wavelength part of the solution indicates an increase in soil moisture or a shallower groundwater table in the repeat survey. Comparison with the processing results of 2D and 3D surveys shows that both image the subsurface, but with local variations which are mainly associated to differences in the acquisition geometry and source types used. Interpretation of baseline and repeat stacks shows that no CO2 related time-lapse signature is observable where the 2D lines allow monitoring of the

  17. Multilayer mounting for long-term light sheet microscopy of zebrafish.

    PubMed

    Weber, Michael; Mickoleit, Michaela; Huisken, Jan

    2014-02-27

    Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is particularly suited for long-term time-lapse imaging over many hours up to several days. However, an appropriate sample mounting strategy is needed that offers both confinement and normal development of the sample. Multilayer mounting, a new embedding technique using low-concentration agarose in optically clear tubes, now overcomes this limitation and unleashes the full potential of light sheet microscopy for real-time developmental biology.

  18. Noise-free accurate count of microbial colonies by time-lapse shadow image analysis.

    PubMed

    Ogawa, Hiroyuki; Nasu, Senshi; Takeshige, Motomu; Funabashi, Hisakage; Saito, Mikako; Matsuoka, Hideaki

    2012-12-01

    Microbial colonies in food matrices could be counted accurately by a novel noise-free method based on time-lapse shadow image analysis. An agar plate containing many clusters of microbial colonies and/or meat fragments was trans-illuminated to project their 2-dimensional (2D) shadow images on a color CCD camera. The 2D shadow images of every cluster distributed within a 3-mm thick agar layer were captured in focus simultaneously by means of a multiple focusing system, and were then converted to 3-dimensional (3D) shadow images. By time-lapse analysis of the 3D shadow images, it was determined whether each cluster comprised single or multiple colonies or a meat fragment. The analytical precision was high enough to be able to distinguish a microbial colony from a meat fragment, to recognize an oval image as two colonies contacting each other, and to detect microbial colonies hidden under a food fragment. The detection of hidden colonies is its outstanding performance in comparison with other systems. The present system attained accuracy for counting fewer than 5 colonies and is therefore of practical importance. Copyright © 2012 Elsevier B.V. All rights reserved.

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

    PubMed Central

    Burke, Russell T.; Orth, James D.

    2016-01-01

    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. PMID:27213923

  20. SSBD: a database of quantitative data of spatiotemporal dynamics of biological phenomena

    PubMed Central

    Tohsato, Yukako; Ho, Kenneth H. L.; Kyoda, Koji; Onami, Shuichi

    2016-01-01

    Motivation: Rapid advances in live-cell imaging analysis and mathematical modeling have produced a large amount of quantitative data on spatiotemporal dynamics of biological objects ranging from molecules to organisms. There is now a crucial need to bring these large amounts of quantitative biological dynamics data together centrally in a coherent and systematic manner. This will facilitate the reuse of this data for further analysis. Results: We have developed the Systems Science of Biological Dynamics database (SSBD) to store and share quantitative biological dynamics data. SSBD currently provides 311 sets of quantitative data for single molecules, nuclei and whole organisms in a wide variety of model organisms from Escherichia coli to Mus musculus. The data are provided in Biological Dynamics Markup Language format and also through a REST API. In addition, SSBD provides 188 sets of time-lapse microscopy images from which the quantitative data were obtained and software tools for data visualization and analysis. Availability and Implementation: SSBD is accessible at http://ssbd.qbic.riken.jp. Contact: sonami@riken.jp PMID:27412095

  1. SSBD: a database of quantitative data of spatiotemporal dynamics of biological phenomena.

    PubMed

    Tohsato, Yukako; Ho, Kenneth H L; Kyoda, Koji; Onami, Shuichi

    2016-11-15

    Rapid advances in live-cell imaging analysis and mathematical modeling have produced a large amount of quantitative data on spatiotemporal dynamics of biological objects ranging from molecules to organisms. There is now a crucial need to bring these large amounts of quantitative biological dynamics data together centrally in a coherent and systematic manner. This will facilitate the reuse of this data for further analysis. We have developed the Systems Science of Biological Dynamics database (SSBD) to store and share quantitative biological dynamics data. SSBD currently provides 311 sets of quantitative data for single molecules, nuclei and whole organisms in a wide variety of model organisms from Escherichia coli to Mus musculus The data are provided in Biological Dynamics Markup Language format and also through a REST API. In addition, SSBD provides 188 sets of time-lapse microscopy images from which the quantitative data were obtained and software tools for data visualization and analysis. SSBD is accessible at http://ssbd.qbic.riken.jp CONTACT: sonami@riken.jp. © The Author 2016. Published by Oxford University Press.

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

    NASA Astrophysics Data System (ADS)

    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

  3. Nuclear apoptotic volume decrease in individual cells: Confocal microscopy imaging and kinetic modeling.

    PubMed

    Khalo, Irina V; Konokhova, Anastasiya I; Orlova, Darya Y; Trusov, Konstantin V; Yurkin, Maxim A; Bartova, Eva; Kozubek, Stanislav; Maltsev, Valeri P; Chernyshev, Andrei V

    2018-05-30

    The dynamics of nuclear morphology changes during apoptosis remains poorly investigated and understood. Using 3D time-lapse confocal microscopy we performed a study of early-stage apoptotic nuclear morphological changes induced by etoposide in single living HepG2 cells. These observations provide a definitive evidence that nuclear apoptotic volume decrease (AVD) is occurring simultaneously with peripheral chromatin condensation (so called "apoptotic ring"). In order to describe quantitatively the dynamics of nuclear morphological changes in the early stage of apoptosis we suggest a general molecular kinetic model, which fits well the obtained experimental data in our study. Results of this work may clarify molecular mechanisms of nuclear morphology changes during apoptosis. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. Investigating glide snow avalanche release using seismic monitoring in combination with time-lapse photography

    NASA Astrophysics Data System (ADS)

    van Herwijnen, Alec; Failletaz, Jerome; Berhod, Nicole; Mitterer, Christoph

    2013-04-01

    Glide avalanches occur when the entire snowpack glides over the ground until an avalanche releases. These avalanches are difficult to forecast since the gliding process can take place over a few hours up to several weeks or months. The presence of liquid water at the interface between the snow cover and the ground surface is of primary importance as it reduces frictional support. Glide avalanches are often preceded by the opening of a tensile crack in the snow cover, called a glide crack. Past research has shown that glide crack opening accelerates prior to avalanche release. During the winter of 2012-2013, we monitored glide crack expansion using time-lapse photography in combination with a seismic sensor and two heat flux sensors on a slope with well documented glide avalanche activity in the Eastern Swiss Alps above Davos, Switzerland. To track changes in glide rates, the number of dark pixels in an area around the glide crack is counted in each image. Using this technique, we observed an increase in glide rates prior to avalanche release. Since the field site is located very close to the town of Davos, the seismic data was very noisy. Nevertheless, the accelerated snow gliding observed in the time-lapse images coincided with increased seismic activity. Overall, these results show that a combination of time-lapse photography with seismic monitoring could provide valuable insight into glide avalanche release. Recordings of the heat flux plates show that the energy input from the soil is fairly small and constant throughout the observed period. The results suggest that ground heat flux is a minor contributor to the water production at the snow-soil interface. Instead, the presence of water at the base of the snowpack is probably due to a strong hydraulic pressure gradient at the snow-soil interface.

  5. Impact of Twitter intensity, time, and location on message lapse of bluebird's pursuit of fleas in Madagascar.

    PubMed

    Da'ar, Omar B; Yunus, Faisel; Md Hossain, Nassif; Househ, Mowafa

    The recent outbreak of bubonic plague in Madagascar reminds us of the continuing public health challenges posed by such deadly diseases in various parts of the world years after their eradication. This study examines the role of Twitter in public health disease surveillance with special focus on how Twitter intensity, time, and location issues explain Twitter plague message delay. We retrospectively analyzed the Twitter feeds of the 2014 bubonic plague outbreak in Madagascar. The analyses are based on the plague-related data available in the public domain between November 19th and 27th 2014. The data were compiled in March 2015. We calculated the time differential between the tweets and retweets, and analyzed various characteristics of the Tweets including Twitter intensity of the users. A total of 6873 Twitter users were included in the study, of which 52% tweeted plague-related information during the morning hours (before mid-day), and 87% of the tweets came from the west of the epicenter of the plague. More importantly, while session of tweet lease and relative location had effect on message lapse, absolute location did not. Additionally, we found no evidence of differential effect of location on message lapse based on relative location i.e. tweets from west or east nor number of following. However, there is evidence that more intense Twitter use appears to have significant effect on message lapse such that as the number of tweets became more intense, time differential between the tweets and retweets increased while higher number of retweets diminished message lapse. This study affirms that Twitter can play an important role in ongoing disease surveillance and the timely dissemination of information during public health emergencies independent of the time and space restrictions. Further ways should be explored to embed social media channels in routine public health practice. Copyright © 2017 King Saud Bin Abdulaziz University for Health Sciences. Published by

  6. Determining absolute protein numbers by quantitative fluorescence microscopy.

    PubMed

    Verdaasdonk, Jolien Suzanne; Lawrimore, Josh; Bloom, Kerry

    2014-01-01

    Biological questions are increasingly being addressed using a wide range of quantitative analytical tools to examine protein complex composition. Knowledge of the absolute number of proteins present provides insights into organization, function, and maintenance and is used in mathematical modeling of complex cellular dynamics. In this chapter, we outline and describe three microscopy-based methods for determining absolute protein numbers--fluorescence correlation spectroscopy, stepwise photobleaching, and ratiometric comparison of fluorescence intensity to known standards. In addition, we discuss the various fluorescently labeled proteins that have been used as standards for both stepwise photobleaching and ratiometric comparison analysis. A detailed procedure for determining absolute protein number by ratiometric comparison is outlined in the second half of this chapter. Counting proteins by quantitative microscopy is a relatively simple yet very powerful analytical tool that will increase our understanding of protein complex composition. © 2014 Elsevier Inc. All rights reserved.

  7. Time-lapse filming of newborn infants. A new technique in monitoring of behavioral states.

    PubMed

    Wallin, A

    1984-05-01

    Knowledge of the newborn infant's behavior is of great importance for an understanding of postnatal adaptation and the influence of exogenous stimuli, such as drugs. To date, however, the methods used have been to sophisticated and complex for use on a busy ward. Moreover, procedures for monitoring physiological parameters in this age group should be non-invasive and gentle for the infant, and acceptable to parents and personnel. In this paper, a simplified technique is presented where observation of the infant can be registered with an ordinary super-8 movie camera adapted to time-lapse. Heart rate, EMG and vocalization are recorded simultaneously. The time-lapse filming technique is relatively easy to learn, it is simple and economic and can be used even during routine clinical care. Assessments were made from the films only, from the recordings only and from a combination of films and recordings. All these modes of assessment were found to be satisfactorily reproducible. Filming combined with respiratory recording gives sufficient information to classify neonatal infant behavior.

  8. Forward modeling to investigate inversion artifacts resulting from time-lapse electrical resistivity tomography during rainfall simulations

    NASA Astrophysics Data System (ADS)

    Carey, Austin M.; Paige, Ginger B.; Carr, Bradley J.; Dogan, Mine

    2017-10-01

    Time-lapse electrical resistivity tomography (ERT) is commonly used as a minimally invasive tool to study infiltration processes. In 2014, we conducted field studies coupling variable intensity rainfall simulation with high-resolution ERT to study the real-time partitioning of rainfall into surface and subsurface response. The significant contrast in resistivity in the subsurface from large changes in subsurface moisture resulted in artifacts during the inversion process of the time-lapse ERT data collected using a dipole-dipole electrode array. These artifacts, which are not representative of real subsurface moisture dynamics, have been shown to arise during time-lapse inversion of ERT data and may be subject to misinterpretation. Forward modeling of the infiltration process post field experiments using a two-layer system (saprolite overlain by a soil layer) was used to generate synthetic datasets. The synthetic data were used to investigate the influence of both changes in volumetric moisture content and electrode configuration on the development of the artifacts identified in the field datasets. For the dipole-dipole array, we found that a decrease in the resistivity of the bottom layer by 67% resulted in a 50% reduction in artifact development. Artifacts for the seven additional array configurations tested, ranged from a 19% increase in artifact development (using an extended dipole-dipole array) to as much as a 96% decrease in artifact development (using a wenner-alpha array), compared to that of the dipole-dipole array. Moreover, these arrays varied in their ability to accurately delineate the infiltration front. Model results showed that the modified pole-dipole array was able to accurately image the infiltration zone and presented fewer artifacts for our experiments. In this study, we identify an optimal array type for imaging rainfall-infiltration dynamics that reduces artifacts. The influence of moisture contrast between the infiltrating water and the

  9. Characteristics of liquid product from the pyrolysis of waste plastic mixture at low and high temperatures: influence of lapse time of reaction.

    PubMed

    Lee, Kyong-Hwan; Shin, Dae-Hyun

    2007-01-01

    Pyrolysis of a waste plastic mixture (high-density polyethylene: low-density polyethylene: polypropylene: polystyrene = 3:2:3:1) into a liquid product was carried out in a stirred semi-batch reactor at low (350 degrees C) and high (400 degrees C) temperatures. The effect of lapse time of reaction in the reactor and also degradation temperature on the characteristics of the liquid product from pyrolysis of the mixture was investigated. Liquid products were described by cumulative amount distribution, paraffin, olefin, naphthene and aromatic (PONA) distribution and molecular weight distribution. Their characteristic was quite differed with a lapse time of reaction and also at a low and high degradation temperatures, because of the different physicochemical properties of the plastic types in the mixture. With increase of lapse time of reaction, the order for the main products in PONA components obtained at 350 degrees C was firstly aromatic products and then olefin products, while at 400 degrees C the order was firstly aromatic products, then olefin products and finally paraffin products. The experiments also showed from the molecular weight distribution of liquid PONA components that the paraffin and olefin products had a wide distribution by mainly random scission of polymer, but in the case of olefin products were produced by an end-chain scission mechanism as well as random scission mechanism, as evidenced by much more light olefin products. This phenomenon was evident at a higher degradation temperature. Also, both the light olefin and naphthene products with a molecular weight of around 120, as a main product, showed a similar trend as a function of lapse time, which had a maximum fraction at 343 min (at 350 degrees C) and 83 min (at 400 degrees C). Among PONA components, the highest concentrations of aromatic products were obtained with a molecular weight of around 100 at the fastest lapse time of reaction, regardless of degradation temperature. It was

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nakanishi, Takeo; Ikenaga, Miho; Fukuda, Hajime

    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 inmore » 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.« less

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

    PubMed

    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.

  12. Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish

    PubMed Central

    Weber, Michael; Mickoleit, Michaela; Huisken, Jan

    2014-01-01

    Light sheet microscopy is the ideal imaging technique to study zebrafish embryonic development. Due to minimal photo-toxicity and bleaching, it is particularly suited for long-term time-lapse imaging over many hours up to several days. However, an appropriate sample mounting strategy is needed that offers both confinement and normal development of the sample. Multilayer mounting, a new embedding technique using low-concentration agarose in optically clear tubes, now overcomes this limitation and unleashes the full potential of light sheet microscopy for real-time developmental biology. PMID:24637614

  13. Integrated time-lapse and single-cell transcription studies highlight the variable and dynamic nature of human hematopoietic cell fate commitment

    PubMed Central

    Moussy, Alice; Cosette, Jérémie; Parmentier, Romuald; da Silva, Cindy; Corre, Guillaume; Richard, Angélique; Gandrillon, Olivier; Stockholm, Daniel

    2017-01-01

    Individual cells take lineage commitment decisions in a way that is not necessarily uniform. We address this issue by characterising transcriptional changes in cord blood-derived CD34+ cells at the single-cell level and integrating data with cell division history and morphological changes determined by time-lapse microscopy. We show that major transcriptional changes leading to a multilineage-primed gene expression state occur very rapidly during the first cell cycle. One of the 2 stable lineage-primed patterns emerges gradually in each cell with variable timing. Some cells reach a stable morphology and molecular phenotype by the end of the first cell cycle and transmit it clonally. Others fluctuate between the 2 phenotypes over several cell cycles. Our analysis highlights the dynamic nature and variable timing of cell fate commitment in hematopoietic cells, links the gene expression pattern to cell morphology, and identifies a new category of cells with fluctuating phenotypic characteristics, demonstrating the complexity of the fate decision process (which is different from a simple binary switch between 2 options, as it is usually envisioned). PMID:28749943

  14. Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography.

    PubMed

    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 < 0.0001) during the course of the observation period. The female pronucleus was significantly smaller in diameter than the male pronucleus (24.1 and 22.4 microm respectively, P = 0.008) and contained fewer nucleoli (4.2 and 7.0 respectively, P < 0.0001). After time-lapse recording, oocytes were cultured for 48 h prior to embryo transfer or cryopreservation. Embryo quality was related to fertilization events and periodicity of the cytoplasmic wave, and it was found that good quality embryos

  15. Monitoring CO 2 Storage at Cranfield, Mississippi with Time-Lapse Offset VSP – Using Integration and Modeling to Reduce Uncertainty

    DOE PAGES

    Daley, Thomas M.; Hendrickson, Joel; Queen, John H.

    2014-12-31

    A time-lapse Offset Vertical Seismic Profile (OVSP) data set was acquired as part of a subsurface monitoring program for geologic sequestration of CO 2. The storage site at Cranfield, near Natchez, Mississippi, is part of a detailed area study (DAS) site for geologic carbon sequestration operated by the U.S. Dept. of Energy’s Southeast Regional Carbon Sequestration Partnership (SECARB). The DAS site includes three boreholes, an injection well and two monitoring wells. The project team selected the DAS site to examine CO 2 sequestration multiphase fluid flow and pressure at the interwell scale in a brine reservoir. The time-lapse (TL) OVSPmore » was part of an integrated monitoring program that included well logs, crosswell seismic, electrical resistance tomography and 4D surface seismic. The goals of the OVSP were to detect the CO 2 induced change in seismic response, give information about the spatial distribution of CO 2 near the injection well and to help tie the high-resolution borehole monitoring to the 4D surface data. The VSP data were acquired in well CFU 31-F1, which is the ~3200 m deep CO 2 injection well at the DAS site. A preinjection survey was recorded in late 2009 with injection beginning in December 2009, and a post injection survey was conducted in Nov 2010 following injection of about 250 kT of CO 2. The sensor array for both surveys was a 50-level, 3-component, Sercel MaxiWave system with 15 m (49 ft) spacing between levels. The source for both surveys was an accelerated weight drop, with different source trucks used for the two surveys. Consistent time-lapse processing was applied to both data sets. Time-lapse processing generated difference corridor stacks to investigate CO 2 induced reflection amplitude changes from each source point. Corridor stacks were used for amplitude analysis to maximize the signal-to-noise ratio (S/N) for each shot point. Spatial variation in reflectivity (used to ‘map’ the plume) was similar in magnitude to

  16. Quantitative Aspects of Single Molecule Microscopy

    PubMed Central

    Ober, Raimund J.; Tahmasbi, Amir; Ram, Sripad; Lin, Zhiping; Ward, E. Sally

    2015-01-01

    Single molecule microscopy is a relatively new optical microscopy technique that allows the detection of individual molecules such as proteins in a cellular context. This technique has generated significant interest among biologists, biophysicists and biochemists, as it holds the promise to provide novel insights into subcellular processes and structures that otherwise cannot be gained through traditional experimental approaches. Single molecule experiments place stringent demands on experimental and algorithmic tools due to the low signal levels and the presence of significant extraneous noise sources. Consequently, this has necessitated the use of advanced statistical signal and image processing techniques for the design and analysis of single molecule experiments. In this tutorial paper, we provide an overview of single molecule microscopy from early works to current applications and challenges. Specific emphasis will be on the quantitative aspects of this imaging modality, in particular single molecule localization and resolvability, which will be discussed from an information theoretic perspective. We review the stochastic framework for image formation, different types of estimation techniques and expressions for the Fisher information matrix. We also discuss several open problems in the field that demand highly non-trivial signal processing algorithms. PMID:26167102

  17. Time-resolved quantitative-phase microscopy of laser-material interactions using a wavefront sensor.

    PubMed

    Gallais, Laurent; Monneret, Serge

    2016-07-15

    We report on a simple and efficient technique based on a wavefront sensor to obtain time-resolved amplitude and phase images of laser-material interactions. The main interest of the technique is to obtain quantitative self-calibrated phase measurements in one shot at the femtosecond time-scale, with high spatial resolution. The technique is used for direct observation and quantitative measurement of the Kerr effect in a fused silica substrate and free electron generation by photo-ionization processes in an optical coating.

  18. High-throughput microfluidics to control and measure signaling dynamics in single yeast cells

    PubMed Central

    Hansen, Anders S.; Hao, Nan; O'Shea, Erin K.

    2015-01-01

    Microfluidics coupled to quantitative time-lapse fluorescence microscopy is transforming our ability to control, measure, and understand signaling dynamics in single living cells. Here we describe a pipeline that incorporates multiplexed microfluidic cell culture, automated programmable fluid handling for cell perturbation, quantitative time-lapse microscopy, and computational analysis of time-lapse movies. We illustrate how this setup can be used to control the nuclear localization of the budding yeast transcription factor Msn2. Using this protocol, we generate oscillations of Msn2 localization and measure the dynamic gene expression response of individual genes in single cells. The protocol allows a single researcher to perform up to 20 different experiments in a single day, whilst collecting data for thousands of single cells. Compared to other protocols, the present protocol is relatively easy to adopt and higher-throughput. The protocol can be widely used to control and monitor single-cell signaling dynamics in other signal transduction systems in microorganisms. PMID:26158443

  19. Time-lapse culture with morphokinetic embryo selection improves pregnancy and live birth chances and reduces early pregnancy loss: a meta-analysis.

    PubMed

    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 < 0.001), significantly lower early pregnancy 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.

  20. Predictors of lapse in first week of smoking abstinence in PTSD and non-PTSD smokers.

    PubMed

    Beckham, Jean C; Calhoun, Patrick S; Dennis, Michelle F; Wilson, Sarah M; Dedert, Eric A

    2013-06-01

    Retrospective research suggests smokers with posttraumatic stress disorder (PTSD) lapse more quickly after their quit date. Ecological momentary assessment (EMA) research is needed to confirm the presence of early smoking lapse in PTSD and form conceptualizations that inform intervention. Smokers with (n = 55) and without (n = 52) PTSD completed alarm-prompted EMA of situational and psychiatric variables the week before and after a quit date, and self-initiated EMA following smoking lapses. Blood samples at baseline and on the quit date allowed assessment of dehydroepiandrosterone (DHEA) and dehydroepiandrosterone sulfate (DHEA(S)). PTSD was related to shorter time to lapse (hazard ratio [HR] = 1.677, 95% CI: 1.106-2.544). Increased smoking abstinence self-efficacy was related to longer time to lapse (HR = 0.608, 95% CI: 0.430-0.860). Analyses of participants' real-time reports revealed that smokers with PTSD were more likely to attribute first-time lapses to negative affect ( = 5.412, p = .020), and trauma reminders (Fisher's exact p = .003**). Finally, the quit date decrease in DHEA(S) was related to shorter time to lapse (HR = 1.009, 95% CI: 1.000-1.018, p < .05). Results provide evidence of shorter time to first smoking lapse in PTSD, and add to evidence that early lapse occasions are more strongly related to trauma reminders, negative affect, and cravings in smokers with PTSD.

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

    PubMed

    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.

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

    PubMed

    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.

  3. Spectral-element simulations of carbon dioxide (CO2) sequestration time-lapse monitoring

    NASA Astrophysics Data System (ADS)

    Morency, C.; Luo, Y.; Tromp, J.

    2009-12-01

    Geologic sequestration of CO2, a green house gas, represents an effort to reduce the large amount of CO2 generated as a by-product of fossil fuels combustion and emitted into the atmosphere. This process of sequestration involves CO2 storage deep underground. There are three main storage options: injection into hydrocarbon reservoirs, injection into methane-bearing coal beds, or injection into deep saline aquifers, that is, highly permeable porous media. The key issues involve accurate monitoring of the CO2, from the injection stage to the prediction & verification of CO2 movement over time for environmental considerations. A natural non-intrusive monitoring technique is referred to as ``4D seismics'', which involves 3D time-lapse seismic surveys. The success of monitoring the CO2 movement is subject to a proper description of the physics of the problem. We propose to realize time-lapse migrations comparing acoustic, elastic, and poroelastic simulations of 4D seismic imaging to characterize the storage zone. This approach highlights the influence of using different physical theories on interpreting seismic data, and, more importantly, on extracting the CO2 signature from the seismic wave field. Our simulations are performed using a spectral-element method, which allows for highly accurate results. Biot's equations are implemented to account for poroelastic effects. Attenuation associated with the anelasticity of the rock frame and frequency-dependent viscous resistance of the pore fluid are accommodated based upon a memory variable approach. The sensitivity of observables to the model parameters is quantified based upon finite-frequency sensitivity kernels calculated using an adjoint method.

  4. A study of possible ``reef effects'' caused by a long-term time-lapse camera in the deep North Pacific

    NASA Astrophysics Data System (ADS)

    Vardaro, M. F.; Parmley, D.; Smith, K. L.

    2007-08-01

    The aggregation response of fish populations following the addition of artificial structures to seafloor habitats has been well documented in shallow-water reefs and at deeper structures such as oil extraction platforms. A long-term time-lapse camera was deployed for 27 four-month deployment periods at 4100 m in the eastern North Pacific to study abyssal megafauna activity and surface-benthos connections. The unique time-series data set provided by this research presented an opportunity to examine how deep-sea benthopelagic fish and epibenthic megafauna populations were affected by an isolated artificial structure and whether animal surveys at this site were biased by aggregation behavior. Counts were taken of benthopelagic grenadiers, Coryphaenoides spp., observed per week as well as numbers of the epibenthic echinoid Echinocrepis rostrata. No significant correlation ( rs=-0.39; p=0.11) was found between the duration of deployment (in weeks) and the average number of Coryphaenoides observed at the site. There was also no evidence of associative behavior around the time-lapse camera by E. rostrata ( rs=-0.32; p=0.19). The results of our study suggest that abyssal fish and epibenthic megafauna do not aggregate around artificial structures and that long-term time-lapse camera studies should not be impacted by aggregation response behaviors.

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

    PubMed

    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.

  6. Isotropic differential phase contrast microscopy for quantitative phase bio-imaging.

    PubMed

    Chen, Hsi-Hsun; Lin, Yu-Zi; Luo, Yuan

    2018-05-16

    Quantitative phase imaging (QPI) has been investigated to retrieve optical phase information of an object and applied to biological microscopy and related medical studies. In recent examples, differential phase contrast (DPC) microscopy can recover phase image of thin sample under multi-axis intensity measurements in wide-field scheme. Unlike conventional DPC, based on theoretical approach under partially coherent condition, we propose a new method to achieve isotropic differential phase contrast (iDPC) with high accuracy and stability for phase recovery in simple and high-speed fashion. The iDPC is simply implemented with a partially coherent microscopy and a programmable thin-film transistor (TFT) shield to digitally modulate structured illumination patterns for QPI. In this article, simulation results show consistency of our theoretical approach for iDPC under partial coherence. In addition, we further demonstrate experiments of quantitative phase images of a standard micro-lens array, as well as label-free live human cell samples. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Quantitative imaging of bilirubin by photoacoustic microscopy

    NASA Astrophysics Data System (ADS)

    Zhou, Yong; Zhang, Chi; Yao, Da-Kang; Wang, Lihong V.

    2013-03-01

    Noninvasive detection of both bilirubin concentration and its distribution is important for disease diagnosis. Here we implemented photoacoustic microscopy (PAM) to detect bilirubin distribution. We first demonstrate that our PAM system can measure the absorption spectra of bilirubin and blood. We also image bilirubin distributions in tissuemimicking samples, both without and with blood mixed. Our results show that PAM has the potential to quantitatively image bilirubin in vivo for clinical applications.

  8. CONFOCAL MICROSCOPY SYSTEM PERFORMANCE: QA TESTS, QUANTITATION AND SPECTROSCOPY

    EPA Science Inventory

    Confocal Microscopy System Performance: QA tests, Quantitation and Spectroscopy.

    Robert M. Zucker 1 and Jeremy M. Lerner 2,
    1Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research Development, U.S. Environmen...

  9. The Extreme Ice Survey: Capturing and Conveying Glacial Processes Through Time-Lapse Imagery and Narration

    NASA Astrophysics Data System (ADS)

    Balog, J. D.; Box, J. E.; Pfeffer, W. T.; Hood, E. W.; Fagre, D. B.; Anker, C.; O'Neel, S.

    2010-12-01

    The Extreme Ice Survey (EIS) uses time-lapse photography, conventional photography, and video to document rapid change in the Earth's glacial ice. The EIS team currently has 38 time-lapse cameras at sites in Greenland, Iceland, Alaska, the Rocky Mountains and Nepal. EIS supplements this ongoing record with annual repeat photography in British Columbia, Iceland, the Alps, and Bolivia. EIS imagery supplies basic knowledge in glacier dynamics to the science community, as well as compelling, engaging narratives to the general public about the immediacy of the Anthropocene and climate change. Visual materials from EIS have impacted more than 150 million people, ranging from White House staff, the U. S. Congress and government agency officials to globally influential corporate officers and all age strata of the general public. Media products include a National Geographic/NOVA special, two National Geographic magazine articles, a feature in Parade magazine (circulation 71 million), and numerous presentations on CNN, NBC, BBC and National Public Radio. Columbia Glacier, Alaska, June 2006, May 2007, June 2008 terminus indicated.

  10. High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy.

    PubMed

    Guo, Baoshan; Lei, Cheng; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

    2017-05-01

    The development of reliable, sustainable, and economical sources of alternative fuels to petroleum is required to tackle the global energy crisis. One such alternative is microalgal biofuel, which is expected to play a key role in reducing the detrimental effects of global warming as microalgae absorb atmospheric CO 2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid amounts and fail to characterize a diverse population of microalgal cells with single-cell resolution in a non-invasive and interference-free manner. Here high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy was demonstrated. In particular, Euglena gracilis, an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement), within lipid droplets was investigated. The optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch quantitative phase microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase maps of every single cell at a high throughput of 10,000 cells/s, enabling accurate cell classification without the need for fluorescent staining. Specifically, the dataset was used to characterize heterogeneous populations of E. gracilis cells under two different culture conditions (nitrogen-sufficient and nitrogen-deficient) and achieve the cell classification with an error rate of only 2.15%. The method holds promise as an effective analytical tool for microalgae-based biofuel production. © 2017 International Society for Advancement of Cytometry. © 2017 International Society for Advancement of Cytometry.

  11. Time-lapse 3-D seismic imaging of shallow subsurface contaminant flow.

    PubMed

    McKenna, J; Sherlock, D; Evans, B

    2001-12-01

    This paper presents a physical modelling study outlining a technique whereby buoyant contaminant flow within water-saturated unconsolidated sand was remotely monitored utilizing the time-lapse 3-D (TL3-D) seismic response. The controlled temperature and pressure conditions, along with the high level of acquisition repeatability attainable using sandbox physical models, allow the TL3-D seismic response to pore fluid movement to be distinguished from all other effects. TL3-D seismic techniques are currently being developed to monitor hydrocarbon reserves within producing reservoirs in an endeavour to improve overall recovery. However, in many ways, sandbox models under atmospheric conditions more accurately simulate the shallow subsurface than petroleum reservoirs. For this reason, perhaps the greatest application for analogue sandbox modelling is to improve our understanding of shallow groundwater and environmental flow mechanisms. Two fluid flow simulations were conducted whereby air and kerosene were injected into separate water-saturated unconsolidated sand models. In both experiments, a base 3-D seismic volume was recorded and compared with six later monitor surveys recorded while the injection program was conducted. Normal incidence amplitude and P-wave velocity information were extracted from the TL3-D seismic data to provide visualization of contaminant migration. Reflection amplitudes displayed qualitative areal distribution of fluids when a suitable impedance contrast existed between pore fluids. TL3-D seismic reflection tomography can potentially monitor the change in areal distribution of fluid contaminants over time, indicating flow patterns. However, other research and this current work have not established a quantifiable relationship between either normal reflection amplitudes and attenuation and fluid saturation. Generally, different pore fluids will have unique seismic velocities due to differences in compressibility and density. The predictable

  12. Molecular and Cellular Quantitative Microscopy: theoretical investigations, technological developments and applications to neurobiology

    NASA Astrophysics Data System (ADS)

    Esposito, Alessandro

    2006-05-01

    This PhD project aims at the development and evaluation of microscopy techniques for the quantitative detection of molecular interactions and cellular features. The primarily investigated techniques are Fαrster Resonance Energy Transfer imaging and Fluorescence Lifetime Imaging Microscopy. These techniques have the capability to quantitatively probe the biochemical environment of fluorophores. An automated microscope capable of unsupervised operation has been developed that enables the investigation of molecular and cellular properties at high throughput levels and the analysis of cellular heterogeneity. State-of-the-art Förster Resonance Energy Transfer imaging, Fluorescence Lifetime Imaging Microscopy, Confocal Laser Scanning Microscopy and the newly developed tools have been combined with cellular and molecular biology techniques for the investigation of protein-protein interactions, oligomerization and post-translational modifications of α-Synuclein and Tau, two proteins involved in Parkinson’s and Alzheimer’s disease, respectively. The high inter-disciplinarity of this project required the merging of the expertise of both the Molecular Biophysics Group at the Debye Institute - Utrecht University and the Cell Biophysics Group at the European Neuroscience Institute - Gαttingen University. This project was conducted also with the support and the collaboration of the Center for the Molecular Physiology of the Brain (Göttingen), particularly with the groups associated with the Molecular Quantitative Microscopy and Parkinson’s Disease and Aggregopathies areas. This work demonstrates that molecular and cellular quantitative microscopy can be used in combination with high-throughput screening as a powerful tool for the investigation of the molecular mechanisms of complex biological phenomena like those occurring in neurodegenerative diseases.

  13. Dynamic phase differences based on quantitative phase imaging for the objective evaluation of cell behavior.

    PubMed

    Krizova, Aneta; Collakova, Jana; Dostal, Zbynek; Kvasnica, Lukas; Uhlirova, Hana; Zikmund, Tomas; Vesely, Pavel; Chmelik, Radim

    2015-01-01

    Quantitative phase imaging (QPI) brought innovation to noninvasive observation of live cell dynamics seen as cell behavior. Unlike the Zernike phase contrast or differential interference contrast, QPI provides quantitative information about cell dry mass distribution. We used such data for objective evaluation of live cell behavioral dynamics by the advanced method of dynamic phase differences (DPDs). The DPDs method is considered a rational instrument offered by QPI. By subtracting the antecedent from the subsequent image in a time-lapse series, only the changes in mass distribution in the cell are detected. The result is either visualized as a two dimensional color-coded projection of these two states of the cell or as a time dependence of changes quantified in picograms. Then in a series of time-lapse recordings, the chain of cell mass distribution changes that would otherwise escape attention is revealed. Consequently, new salient features of live cell behavior should emerge. Construction of the DPDs method and results exhibiting the approach are presented. Advantage of the DPDs application is demonstrated on cells exposed to an osmotic challenge. For time-lapse acquisition of quantitative phase images, the recently developed coherence-controlled holographic microscope was employed.

  14. Dynamic phase differences based on quantitative phase imaging for the objective evaluation of cell behavior

    NASA Astrophysics Data System (ADS)

    Krizova, Aneta; Collakova, Jana; Dostal, Zbynek; Kvasnica, Lukas; Uhlirova, Hana; Zikmund, Tomas; Vesely, Pavel; Chmelik, Radim

    2015-11-01

    Quantitative phase imaging (QPI) brought innovation to noninvasive observation of live cell dynamics seen as cell behavior. Unlike the Zernike phase contrast or differential interference contrast, QPI provides quantitative information about cell dry mass distribution. We used such data for objective evaluation of live cell behavioral dynamics by the advanced method of dynamic phase differences (DPDs). The DPDs method is considered a rational instrument offered by QPI. By subtracting the antecedent from the subsequent image in a time-lapse series, only the changes in mass distribution in the cell are detected. The result is either visualized as a two-dimensional color-coded projection of these two states of the cell or as a time dependence of changes quantified in picograms. Then in a series of time-lapse recordings, the chain of cell mass distribution changes that would otherwise escape attention is revealed. Consequently, new salient features of live cell behavior should emerge. Construction of the DPDs method and results exhibiting the approach are presented. Advantage of the DPDs application is demonstrated on cells exposed to an osmotic challenge. For time-lapse acquisition of quantitative phase images, the recently developed coherence-controlled holographic microscope was employed.

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

    PubMed

    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.

  16. Relations among affect, abstinence motivation and confidence, and daily smoking lapse risk.

    PubMed

    Minami, Haruka; Yeh, Vivian M; Bold, Krysten W; Chapman, Gretchen B; McCarthy, Danielle E

    2014-06-01

    This study tested the hypothesis that changes in momentary affect, abstinence motivation, and confidence would predict lapse risk over the next 12-24 hr using Ecological Momentary Assessment (EMA) data from smokers attempting to quit smoking. One hundred and three adult, daily, treatment-seeking smokers recorded their momentary affect, motivation to quit, abstinence confidence, and smoking behaviors in near real time with multiple EMA reports per day using electronic diaries postquit. Multilevel models indicated that initial levels of negative affect were associated with smoking, even after controlling for earlier smoking status, and that short-term increases in negative affect predicted lapses up to 12, but not 24, hr later. Positive affect had significant effects on subsequent abstinence confidence, but not motivation to quit. High levels of motivation appeared to reduce increases in lapse risk that occur over hours although momentary changes in confidence did not predict lapse risk over 12 hr. Negative affect had short-lived effects on lapse risk, whereas higher levels of motivation protected against the risk of lapsing that accumulates over hours. An increase in positive affect was associated with greater confidence to quit, but such changes in confidence did not reduce short-term lapse risk, contrary to expectations. Relations observed among affect, cognitions, and lapse seem to depend critically on the timing of assessments.

  17. Relations among Affect, Abstinence Motivation and Confidence, and Daily Smoking Lapse Risk

    PubMed Central

    Minami, Haruka; Yeh, Vivian M.; Bold, Krysten W.; Chapman, Gretchen B.; McCarthy, Danielle E.

    2016-01-01

    Aims This study tested the hypothesis that changes in momentary affect, abstinence motivation, and confidence would predict lapse risk over the next 12–24 hours using Ecological Momentary Assessment (EMA) data from smokers attempting to quit smoking. Method 103 adult, daily, treatment-seeking smokers recorded their momentary affect, motivation to quit, abstinence confidence, and smoking behaviors in near real time with multiple EMA reports per day using electronic diaries post-quit. Results Multilevel models indicated that initial levels of negative affect were associated with smoking, even after controlling for earlier smoking status, and that short-term increases in negative affect predicted lapses up to 12, but not 24, hours later. Positive affect had significant effects on subsequent abstinence confidence, but not motivation to quit. High levels of motivation appeared to reduce increases in lapse risk that occur over hours while momentary changes in confidence did not predict lapse risk over 12 hours. Conclusion Negative affect had short-lived effects on lapse risk, whereas higher levels of motivation protected against the risk of lapsing that accumulates over hours. An increase in positive affect was associated with greater confidence to quit, but such changes in confidence did not reduce short-term lapse risk, contrary to expectations. Relations observed among affect, cognitions, and lapse seem to depend critically on the timing of assessments. PMID:24955665

  18. A Simple Configuration for Quantitative Phase Contrast Microscopy of Transmissible Samples

    NASA Astrophysics Data System (ADS)

    Sengupta, Chandan; Dasgupta, Koustav; Bhattacharya, K.

    Phase microscopy attempts to visualize and quantify the phase distribution of samples which are otherwise invisible under microscope without the use of stains. The two principal approaches to phase microscopy are essentially those of Fourier plane modulation and interferometric techniques. Although the former, first proposed by Zernike, had been the harbinger of phase microscopy, it was the latter that allowed for quantitative evaluation of phase samples. However interferometric techniques are fraught with associated problems such as complicated setup involving mirrors and beam-splitters, the need for a matched objective in the reference arm and also the need for vibration isolation. The present work proposes a single element cube beam-splitter (CBS) interferometer combined with a microscope objective (MO) for interference microscopy. Because of the monolithic nature of the interferometer, the system is almost insensitive to vibrations and relatively simple to align. It will be shown that phase shifting properties may also be introduced by suitable and proper use of polarizing devices. Initial results showing the quantitative three dimensional phase profiles of simulated and actual biological specimens are presented.

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

    PubMed Central

    Farooq, Muhammad; Kim, Jung Ho; Song, Young Soo; Amjad Sabir, Mohammad; Umar, Muhammad; Tariq, Mohammad; Muhammad, Said

    2014-01-01

    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. PMID:24578621

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

    NASA Astrophysics Data System (ADS)

    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.

  1. Time-lapse electrical resistivity investigations for imaging the grouting injection in shallow subsurface cavities.

    PubMed

    Farooq, Muhammad; Park, Samgyu; Kim, Jung Ho; Song, Young Soo; Amjad Sabir, Mohammad; Umar, Muhammad; Tariq, Mohammad; Muhammad, Said

    2014-01-01

    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.

  2. Assessing Uncertainty and Repeatability in Time-Lapse VSP Monitoring of CO2 Injection in a Brine Aquifer, Frio Formation, Texas (A Case Study)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nazari, Siamak; Daley, Thomas M.

    2013-02-07

    This study was done to assess the repeatability and uncertainty of time-lapse VSP response to CO 2 injection in the Frio formation near Houston Texas. A work flow was built to assess the effect of time-lapse injected CO 2 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 formore » 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 of VSP for CO

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

    USGS Publications Warehouse

    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.

  4. A versatile soft X-ray transmission system for time resolved in situ microscopy with chemical contrast.

    PubMed

    Forsberg, J; Englund, C-J; Duda, L-C

    2009-08-01

    We present the design and operation of a versatile soft X-ray transmission system for time resolved in situ microscopy with chemical contrast. The utility of the setup is demonstrated by results from following a corrosion process of iron in saline environment, subjected to a controlled humid atmosphere. The system includes a transmission flow-cell reactor that allows for in situ microscopic probing with soft X-rays. We employ a full field technique by using a nearly collimated X-ray beam that produces an unmagnified projection of the transmitted soft X-rays (below 1.1 keV) which is magnified and recorded by an optical CCD camera. Time lapse series with chemical contrast allow us to follow and interpret the chemical processes in detail. The obtainable lateral resolution is a few mum, sufficient to detect filiform corrosion on iron.

  5. Time-Lapse Monitoring with 4D Seismic Coda Waves in Active, Passive and Ambient Noise Data

    NASA Astrophysics Data System (ADS)

    Lumley, D. E.; Kamei, R.; Saygin, E.; Shragge, J. C.

    2017-12-01

    The Earth's subsurface is continuously changing, due to temporal variations in fluid flow, stress, temperature, geomechanics and geochemistry, for example. These physical changes occur at broad tectonic and earthquake scales, and also at very detailed near-surface and reservoir scales. Changes in the physical states of the earth cause time-varying changes in the physical properties of rocks and fluids, which can be monitored with natural or manmade seismic waves. Time-lapse (4D) seismic monitoring is important for applications related to natural and induced seismicity, hydrocarbon and groundwater reservoir depletion, CO2 sequestration etc. An exciting new research area involves moving beyond traditional methods in order to use the full complex time-lapse scattered wavefield (4D coda waves) for both manmade active-source 3D/4D seismic data, and also to use continuous recordings of natural-source passive seismic data, especially (micro) earthquakes and ocean ambient noise. This research involves full wave-equation approaches including waveform inversion (FWI), interferometry, Large N sensor arrays, "big data" information theory, and high performance supercomputing (HPC). I will present high-level concepts and recent data results that are quite spectacular and highly encouraging.

  6. Recreational use assessment of water-based activities, using time-lapse construction cameras.

    PubMed

    Sunger, Neha; Teske, Sondra S; Nappier, Sharon; Haas, Charles N

    2012-01-01

    Recreational exposure to surface waters during periods of increased pathogen concentration may lead to a significantly higher risk of illness. However, estimates of elementary exposure factors necessary to evaluate health risk (i.e., usage distributions and exposure durations) are not available for many non-swimming water-related activities. No prior studies have assessed non-swimming water exposure with respect to factors leading to impaired water quality from increased pathogen concentration, such as weather condition (rain events produce increased runoff and sewer overflows) and type of day (heavy recreational periods). We measured usage patterns and evaluated the effect of weather and type of day at eight water sites located within Philadelphia, by using a novel "time lapse photography" technology during three peak recreational seasons (May-September) 2008-2010. Camera observations validated with simultaneous in-person surveys exhibited a strong correlation (R(2)=0.81 to 0.96) between the two survey techniques, indicating that the application of remote photography in collecting human exposure data was appropriate. Recreational activities usage varied more on a temporal basis than due to inclement weather. Only 14% (6 out of 44) of the site-specific activity combinations showed dry weather preference, whereas 41.5% (17 out of 41) of the combinations indicated greater usage on weekends as compared with weekday. In general, the log normal distribution described the playing and wading duration distribution, while the gamma distribution was the best fit for fishing durations. Remote photography provided unbiased, real-time human exposure data and was less personnel intensive compared with traditional survey methods. However, there are potential limitations associated with remote surveillance data related to its limited view. This is the first study to report that time lapse cameras can be successfully applied to assess water-based human recreational patterns and can

  7. Oil Sands Characteristics and Time-Lapse and P-SV Seismic Steam Monitoring, Athabasca, Canada

    NASA Astrophysics Data System (ADS)

    Takahashi, A.; Nakayama, T.; Kashihara, K.; Skinner, L.; Kato, A.

    2008-12-01

    -injection. The differences of the seismic responses between the time-lapse seismic volumes can be quantitatively explained by P-wave velocity decrease of the oil sands layers due to steam-injection. In addition, the data suggests that a larger area would be influenced by pressure than temperature. We calculate several seismic attributes such as RMS values of amplitude difference, maximum cross correlations, and interval velocity differences. These attributes are integrated by using self-organization maps (SOM) and K-means methods. By this analysis, we are able to distinguish areas of steam chamber growth from transitional and non-affected areas. In addition, 3D P-SV converted-wave processing and analysis are applied on the second 3D data set (recorded with three-component digital sensor). Low Vp/Vs values in the P-SV volume show areas of steam chamber development, and high Vp/Vs values indicate transitional zones. Our analysis of both time-lapse 3D seismic and 3D P-SV data along with the rock physics model can be used to monitor qualitatively and quantitatively the rock property changes of the inter-well reservoir sands in the field.

  8. Lapse-time-dependent coda-wave depth sensitivity to local velocity perturbations in 3-D heterogeneous elastic media

    NASA Astrophysics Data System (ADS)

    Obermann, Anne; Planès, Thomas; Hadziioannou, Céline; Campillo, Michel

    2016-10-01

    In the context of seismic monitoring, recent studies made successful use of seismic coda waves to locate medium changes on the horizontal plane. Locating the depth of the changes, however, remains a challenge. In this paper, we use 3-D wavefield simulations to address two problems: first, we evaluate the contribution of surface- and body-wave sensitivity to a change at depth. We introduce a thin layer with a perturbed velocity at different depths and measure the apparent relative velocity changes due to this layer at different times in the coda and for different degrees of heterogeneity of the model. We show that the depth sensitivity can be modelled as a linear combination of body- and surface-wave sensitivity. The lapse-time-dependent sensitivity ratio of body waves and surface waves can be used to build 3-D sensitivity kernels for imaging purposes. Second, we compare the lapse-time behaviour in the presence of a perturbation in horizontal and vertical slabs to address, for instance, the origin of the velocity changes detected after large earthquakes.

  9. Cytoskeleton-dependent transport of cytoplasmic particles in previtellogenic to mid-vitellogenic ovarian follicles of Drosophila: time-lapse analysis using video-enhanced contrast microscopy.

    PubMed

    Bohrmann, J; Biber, K

    1994-04-01

    In Drosophila oogenesis, several morphogenetic determinants and other developmental factors synthesized in the nurse cells have been shown to accumulate in the oocyte during pre- to mid-vitellogenic stages. However, the mechanisms of the involved intercellular transport processes that seem to be rather selective have not been revealed so far. We have investigated in vitro, by means of video-enhanced contrast time-lapse microscopy, the transport of cytoplasmic particles from the nurse cells through ring canals into the oocyte during oogenesis stages 6-10A. At stage 7, we first observed single particles moving into the previtellogenic oocyte. The particle transfer was strictly unidirectional and seemed to be selective, since only some individual particles moved whereas other particles lying in the vicinity of the ring canals were not transported. The observed transport processes were inhibitable with 2,4-dinitrophenol, cytochalasin B or N-ethylmaleimide, but not with microtubule inhibitors. At the beginning of vitellogenesis (stage 8), the selective translocation of particles through the ring canals became faster (up to 130 nm/second) and more frequent (about 1 particle/minute), whereas during mid-vitellogenesis (stages 9-10A) the velocity and the frequency of particle transport decreased again. Following their more or less rectilinear passage through the ring canals, the particles joined a circular stream of cytoplasmic particles in the oocyte. This ooplasmic particle streaming started at stage 6/7 with velocities of about 80 nm/second and some reversals of direction at the beginning. The particle stream in the oocyte was sensitive to colchicine and vinblastine, but not to cytochalasin B, and we presume that it reflects the rearrangement of ooplasmic microtubules described recently by other authors. We propose that during stages 7-10A, a selective transport of particles into the oocyte occurs through the ring canal along a polarized scaffold of cytoskeletal elements

  10. Terahertz time-lapse imaging of hydration in physiological tissues

    NASA Astrophysics Data System (ADS)

    Bennett, David B.; Taylor, Zachary D.; Bajwa, Neha; Tewari, Priyamvada; Maccabi, Ashkan; Sung, Shijun; Singh, Rahul S.; Culjat, Martin O.; Grundfest, Warren S.; Brown, Elliott R.

    2011-02-01

    This study describes terahertz (THz) imaging of hydration changes in physiological tissues with high water concentration sensitivity. A fast-scanning, pulsed THz imaging system (centered at 525 GHz; 125 GHz bandwidth) was utilized to acquire a 35 mm x 35 mm field-of-view with 0.5 mm x 0.5 mm pixels in less than two minutes. THz time-lapsed images were taken on three sample systems: (1) a simple binary system of water evaporating from a polypropylene towel, (2) the accumulation of fluid at the site of a sulfuric acid burn on ex vivo porcine skin, and (3) the evaporative dehydration of an ex vivo porcine cornea. The diffusion-regulating behavior of corneal tissue is elucidated, and the correlation of THz reflectivity with tissue hydration is measured using THz spectroscopy on four ex vivo corneas. We conclude that THz imaging can discern small differences in the distribution of water in physiological tissues and is a good candidate for burn and corneal imaging.

  11. Application of Machine Learning to Predict Dietary Lapses During Weight Loss.

    PubMed

    Goldstein, Stephanie P; Zhang, Fengqing; Thomas, John G; Butryn, Meghan L; Herbert, James D; Forman, Evan M

    2018-05-01

    Individuals who adhere to dietary guidelines provided during weight loss interventions tend to be more successful with weight control. Any deviation from dietary guidelines can be referred to as a "lapse." There is a growing body of research showing that lapses are predictable using a variety of physiological, environmental, and psychological indicators. With recent technological advancements, it may be possible to assess these triggers and predict dietary lapses in real time. The current study sought to use machine learning techniques to predict lapses and evaluate the utility of combining both group- and individual-level data to enhance lapse prediction. The current study trained and tested a machine learning algorithm capable of predicting dietary lapses from a behavioral weight loss program among adults with overweight/obesity (n = 12). Participants were asked to follow a weight control diet for 6 weeks and complete ecological momentary assessment (EMA; repeated brief surveys delivered via smartphone) regarding dietary lapses and relevant triggers. WEKA decision trees were used to predict lapses with an accuracy of 0.72 for the group of participants. However, generalization of the group algorithm to each individual was poor, and as such, group- and individual-level data were combined to improve prediction. The findings suggest that 4 weeks of individual data collection is recommended to attain optimal model performance. The predictive algorithm could be utilized to provide in-the-moment interventions to prevent dietary lapses and therefore enhance weight losses. Furthermore, methods in the current study could be translated to other types of health behavior lapses.

  12. Time-lapse resistivity investigation of salinity changes at an ex-promontory land: a case study of Carey Island, Selangor, Malaysia.

    PubMed

    Tajul Baharuddin, Mohamad Faizal; Taib, Samsudin; Hashim, Roslan; Zainal Abidin, Mohd Hazreek; Ishak, Mohd Fakhrurrazi

    2011-09-01

    Time-lapse resistivity measurements and groundwater geochemistry were used to study salinity effect on groundwater aquifer at the ex-promontory-land of Carey Island in Malaysia. Resistivity was measured by ABEM Terrameter SAS4000 and ES10-64 electrode selector. Relationship between earth resistivity and total dissolved solids (TDS) was derived, and with resistivity images, used to identify water types: fresh (ρ ( e ) > 6.5 Ω m), brackish (3 Ω m < ρ ( e ) < 6.5 Ω m), or saline (ρ ( e ) < 3 Ω m). Long-term monitoring of the studied area's groundwater quality via measurements of its time-lapse resistivity showed salinity changes in the island's groundwater aquifers not conforming to seawater-freshwater hydraulic gradient. In some aquifers far from the coast, saline water was dominant, while in some others, freshwater 30 m thick showed groundwater potential. Land transformation is believed to have changed the island's hydrogeology, which receives saltwater pressure all the time, limiting freshwater recharge to the groundwater system. The time-lapse resistivity measurements showed active salinity changes at resistivity-image bottom moving up the image for two seasons' (wet and dry) conditions. The salinity changes are believed to have been caused by incremental tide passing through highly porous material in the active-salinity-change area. The study's results were used to plan a strategy for sustainable groundwater exploration of the island.

  13. Scanning fluorescent microscopy is an alternative for quantitative fluorescent cell analysis.

    PubMed

    Varga, Viktor Sebestyén; Bocsi, József; Sipos, Ferenc; Csendes, Gábor; Tulassay, Zsolt; Molnár, Béla

    2004-07-01

    Fluorescent measurements on cells are performed today with FCM and laser scanning cytometry. The scientific community dealing with quantitative cell analysis would benefit from the development of a new digital multichannel and virtual microscopy based scanning fluorescent microscopy technology and from its evaluation on routine standardized fluorescent beads and clinical specimens. We applied a commercial motorized fluorescent microscope system. The scanning was done at 20 x (0.5 NA) magnification, on three channels (Rhodamine, FITC, Hoechst). The SFM (scanning fluorescent microscopy) software included the following features: scanning area, exposure time, and channel definition, autofocused scanning, densitometric and morphometric cellular feature determination, gating on scatterplots and frequency histograms, and preparation of galleries of the gated cells. For the calibration and standardization Immuno-Brite beads were used. With application of shading compensation, the CV of fluorescence of the beads decreased from 24.3% to 3.9%. Standard JPEG image compression until 1:150 resulted in no significant change. The change of focus influenced the CV significantly only after +/-5 microm error. SFM is a valuable method for the evaluation of fluorescently labeled cells. Copyright 2004 Wiley-Liss, Inc.

  14. DMD-based quantitative phase microscopy and optical diffraction tomography

    NASA Astrophysics Data System (ADS)

    Zhou, Renjie

    2018-02-01

    Digital micromirror devices (DMDs), which offer high speed and high degree of freedoms in steering light illuminations, have been increasingly applied to optical microscopy systems in recent years. Lately, we introduced DMDs into digital holography to enable new imaging modalities and break existing imaging limitations. In this paper, we will first present our progress in using DMDs for demonstrating laser-illumination Fourier ptychographic microscopy (FPM) with shotnoise limited detection. After that, we will present a novel common-path quantitative phase microscopy (QPM) system based on using a DMD. Building on those early developments, a DMD-based high speed optical diffraction tomography (ODT) system has been recently demonstrated, and the results will also be presented. This ODT system is able to achieve video-rate 3D refractive-index imaging, which can potentially enable observations of high-speed 3D sample structural changes.

  15. Improved cancer risk stratification and diagnosis via quantitative phase microscopy (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Liu, Yang; Uttam, Shikhar; Pham, Hoa V.; Hartman, Douglas J.

    2017-02-01

    Pathology remains the gold standard for cancer diagnosis and in some cases prognosis, in which trained pathologists examine abnormality in tissue architecture and cell morphology characteristic of cancer cells with a bright-field microscope. The limited resolution of conventional microscope can result in intra-observer variation, missed early-stage cancers, and indeterminate cases that often result in unnecessary invasive procedures in the absence of cancer. Assessment of nanoscale structural characteristics via quantitative phase represents a promising strategy for identifying pre-cancerous or cancerous cells, due to its nanoscale sensitivity to optical path length, simple sample preparation (i.e., label-free) and low cost. I will present the development of quantitative phase microscopy system in transmission and reflection configuration to detect the structural changes in nuclear architecture, not be easily identifiable by conventional pathology. Specifically, we will present the use of transmission-mode quantitative phase imaging to improve diagnostic accuracy of urine cytology and the nuclear dry mass is progressively correlate with negative, atypical, suspicious and positive cytological diagnosis. In a second application, we will present the use of reflection-mode quantitative phase microscopy for depth-resolved nanoscale nuclear architecture mapping (nanoNAM) of clinically prepared formalin-fixed, paraffin-embedded tissue sections. We demonstrated that the quantitative phase microscopy system detects a gradual increase in the density alteration of nuclear architecture during malignant transformation in animal models of colon carcinogenesis and in human patients with ulcerative colitis, even in tissue that appears histologically normal according to pathologists. We evaluated the ability of nanoNAM to predict "future" cancer progression in patients with ulcerative colitis.

  16. A Dynamic Programming Model for Optimizing Frequency of Time-Lapse Seismic Monitoring in Geological CO2 Storage

    NASA Astrophysics Data System (ADS)

    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

  17. Time-lapse contact microscopy of cell cultures based on non-coherent illumination

    NASA Astrophysics Data System (ADS)

    Gabriel, Marion; Balle, Dorothée; Bigault, Stéphanie; Pornin, Cyrille; Gétin, Stéphane; Perraut, François; Block, Marc R.; Chatelain, François; Picollet-D'Hahan, Nathalie; Gidrol, Xavier; Haguet, Vincent

    2015-10-01

    Video microscopy offers outstanding capabilities to investigate the dynamics of biological and pathological mechanisms in optimal culture conditions. Contact imaging is one of the simplest imaging architectures to digitally record images of cells due to the absence of any objective between the sample and the image sensor. However, in the framework of in-line holography, other optical components, e.g., an optical filter or a pinhole, are placed underneath the light source in order to illuminate the cells with a coherent or quasi-coherent incident light. In this study, we demonstrate that contact imaging with an incident light of both limited temporal and spatial coherences can be achieved with sufficiently high quality for most applications in cell biology, including monitoring of cell sedimentation, rolling, adhesion, spreading, proliferation, motility, death and detachment. Patterns of cells were recorded at various distances between 0 and 1000 μm from the pixel array of the image sensors. Cells in suspension, just deposited or at mitosis focalise light into photonic nanojets which can be visualised by contact imaging. Light refraction by cells significantly varies during the adhesion process, the cell cycle and among the cell population in connection with every modification in the tridimensional morphology of a cell.

  18. Calibration of Wide-Field Deconvolution Microscopy for Quantitative Fluorescence Imaging

    PubMed Central

    Lee, Ji-Sook; Wee, Tse-Luen (Erika); Brown, Claire M.

    2014-01-01

    Deconvolution enhances contrast in fluorescence microscopy images, especially in low-contrast, high-background wide-field microscope images, improving characterization of features within the sample. Deconvolution can also be combined with other imaging modalities, such as confocal microscopy, and most software programs seek to improve resolution as well as contrast. Quantitative image analyses require instrument calibration and with deconvolution, necessitate that this process itself preserves the relative quantitative relationships between fluorescence intensities. To ensure that the quantitative nature of the data remains unaltered, deconvolution algorithms need to be tested thoroughly. This study investigated whether the deconvolution algorithms in AutoQuant X3 preserve relative quantitative intensity data. InSpeck Green calibration microspheres were prepared for imaging, z-stacks were collected using a wide-field microscope, and the images were deconvolved using the iterative deconvolution algorithms with default settings. Afterwards, the mean intensities and volumes of microspheres in the original and the deconvolved images were measured. Deconvolved data sets showed higher average microsphere intensities and smaller volumes than the original wide-field data sets. In original and deconvolved data sets, intensity means showed linear relationships with the relative microsphere intensities given by the manufacturer. Importantly, upon normalization, the trend lines were found to have similar slopes. In original and deconvolved images, the volumes of the microspheres were quite uniform for all relative microsphere intensities. We were able to show that AutoQuant X3 deconvolution software data are quantitative. In general, the protocol presented can be used to calibrate any fluorescence microscope or image processing and analysis procedure. PMID:24688321

  19. In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

    PubMed Central

    Malide, Daniela; Métais, Jean-Yves; Dunbar, Cynthia E.

    2014-01-01

    We developed and validated a fluorescent marking methodology for clonal tracking of hematopoietic stem and progenitor cells (HSPCs) with high spatial and temporal resolution to study in vivo hematopoiesis using the murine bone marrow transplant experimental model. Genetic combinatorial marking using lentiviral vectors encoding fluorescent proteins (FPs) enabled cell fate mapping through advanced microscopy imaging. Vectors encoding five different FPs: Cerulean, EGFP, Venus, tdTomato, and mCherry were used to concurrently transduce HSPCs, creating a diverse palette of color marked cells. Imaging using confocal/two-photon hybrid microscopy enables simultaneous high resolution assessment of uniquely marked cells and their progeny in conjunction with structural components of the tissues. Volumetric analyses over large areas reveal that spectrally coded HSPC-derived cells can be detected non-invasively in various intact tissues, including the bone marrow (BM), for extensive periods of time following transplantation. Live studies combining video-rate multiphoton and confocal time-lapse imaging in 4D demonstrate the possibility of dynamic cellular and clonal tracking in a quantitative manner. PMID:25145579

  20. HAADF-STEM atom counting in atom probe tomography specimens: Towards quantitative correlative microscopy.

    PubMed

    Lefebvre, W; Hernandez-Maldonado, D; Moyon, F; Cuvilly, F; Vaudolon, C; Shinde, D; Vurpillot, F

    2015-12-01

    The geometry of atom probe tomography tips strongly differs from standard scanning transmission electron microscopy foils. Whereas the later are rather flat and thin (<20 nm), tips display a curved surface and a significantly larger thickness. As far as a correlative approach aims at analysing the same specimen by both techniques, it is mandatory to explore the limits and advantages imposed by the particular geometry of atom probe tomography specimens. Based on simulations (electron probe propagation and image simulations), the possibility to apply quantitative high angle annular dark field scanning transmission electron microscopy to of atom probe tomography specimens has been tested. The influence of electron probe convergence and the benefice of deconvolution of electron probe point spread function electron have been established. Atom counting in atom probe tomography specimens is for the first time reported in this present work. It is demonstrated that, based on single projections of high angle annular dark field imaging, significant quantitative information can be used as additional input for refining the data obtained by correlative analysis of the specimen in APT, therefore opening new perspectives in the field of atomic scale tomography. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. High-Precision Pinpointing of Luminescent Targets in Encoder-Assisted Scanning Microscopy Allowing High-Speed Quantitative Analysis.

    PubMed

    Zheng, Xianlin; Lu, Yiqing; Zhao, Jiangbo; Zhang, Yuhai; Ren, Wei; Liu, Deming; Lu, Jie; Piper, James A; Leif, Robert C; Liu, Xiaogang; Jin, Dayong

    2016-01-19

    Compared with routine microscopy imaging of a few analytes at a time, rapid scanning through the whole sample area of a microscope slide to locate every single target object offers many advantages in terms of simplicity, speed, throughput, and potential for robust quantitative analysis. Existing techniques that accommodate solid-phase samples incorporating individual micrometer-sized targets generally rely on digital microscopy and image analysis, with intrinsically low throughput and reliability. Here, we report an advanced on-the-fly stage scanning method to achieve high-precision target location across the whole slide. By integrating X- and Y-axis linear encoders to a motorized stage as the virtual "grids" that provide real-time positional references, we demonstrate an orthogonal scanning automated microscopy (OSAM) technique which can search a coverslip area of 50 × 24 mm(2) in just 5.3 min and locate individual 15 μm lanthanide luminescent microspheres with standard deviations of 1.38 and 1.75 μm in X and Y directions. Alongside implementation of an autofocus unit that compensates the tilt of a slide in the Z-axis in real time, we increase the luminescence detection efficiency by 35% with an improved coefficient of variation. We demonstrate the capability of advanced OSAM for robust quantification of luminescence intensities and lifetimes for a variety of micrometer-scale luminescent targets, specifically single down-shifting and upconversion microspheres, crystalline microplates, and color-barcoded microrods, as well as quantitative suspension array assays of biotinylated-DNA functionalized upconversion nanoparticles.

  2. Are seismic velocity time-lapse changes due to fluid substitution or matrix dissolution? A CO2 sequestration study at Pohokura Field, New Zealand

    NASA Astrophysics Data System (ADS)

    Adam, L.; Sim, C. Y.; Macfarlane, J.; van Wijk, K.; Shragge, J. C.; Higgs, K.

    2015-12-01

    Time-lapse seismic signatures can be used to quantify fluid saturation and pressure changes in a reservoir undergoing CO2 sequestration. However, the injection of CO2 acidifies the water, which may dissolve and/or precipitate minerals. Understanding the impact on the rock frame from field seismic time-lapse changes remains an outstanding challenge. Here, we study the effects of carbonate-CO2-water reactions on the physical and elastic properties of rock samples with variable volumes of carbonate cementation. The effects of fluid substitution alone (brine to CO2) and those due to the combination of fluid substitution and mineral dissolution on time-lapse seismic signatures are studied by combining laboratory data, geophysical well-log data and 1-D seismic modeling. Nine rocks from Pohokura Field (New Zealand) are reacted with carbonic acid. The elastic properties are measured using a high-density laser-ultrasonic setup. We observe that P-wave velocity changes up to -19% and correlate with sandstone grain size. Coarse-grained sandstones show greater changes in elastic wave velocities due to dissolution than fine-grained sandstones. To put this in perspective, this velocity change is comparable to the effect of fluid substitution from brine to CO2. This can potentially create an ambiguity in the interpretation of the physical processes responsible for time-lapse signatures in a CO2injection scenario. The laboratory information is applied onto well-log data to model changes in elastic properties of sandstones at the well-log scale. Well-logs and core petrographic analyses are used to find an elastic model that best describes the observed elastic waves velocities in the cemented reservoir sandstones. The Constant-cement rock physics model is found to predict the elastic behaviour of the cemented sandstones. A possible late-time sequestration scenario is that both mineral dissolution and fluid substitution occur in the reservoir. 1-D synthetic seismograms show that

  3. Image correlation microscopy for uniform illumination.

    PubMed

    Gaborski, T R; Sealander, M N; Ehrenberg, M; Waugh, R E; McGrath, J L

    2010-01-01

    Image cross-correlation microscopy is a technique that quantifies the motion of fluorescent features in an image by measuring the temporal autocorrelation function decay in a time-lapse image sequence. Image cross-correlation microscopy has traditionally employed laser-scanning microscopes because the technique emerged as an extension of laser-based fluorescence correlation spectroscopy. In this work, we show that image correlation can also be used to measure fluorescence dynamics in uniform illumination or wide-field imaging systems and we call our new approach uniform illumination image correlation microscopy. Wide-field microscopy is not only a simpler, less expensive imaging modality, but it offers the capability of greater temporal resolution over laser-scanning systems. In traditional laser-scanning image cross-correlation microscopy, lateral mobility is calculated from the temporal de-correlation of an image, where the characteristic length is the illuminating laser beam width. In wide-field microscopy, the diffusion length is defined by the feature size using the spatial autocorrelation function. Correlation function decay in time occurs as an object diffuses from its original position. We show that theoretical and simulated comparisons between Gaussian and uniform features indicate the temporal autocorrelation function depends strongly on particle size and not particle shape. In this report, we establish the relationships between the spatial autocorrelation function feature size, temporal autocorrelation function characteristic time and the diffusion coefficient for uniform illumination image correlation microscopy using analytical, Monte Carlo and experimental validation with particle tracking algorithms. Additionally, we demonstrate uniform illumination image correlation microscopy analysis of adhesion molecule domain aggregation and diffusion on the surface of human neutrophils.

  4. Automated quantitative cytological analysis using portable microfluidic microscopy.

    PubMed

    Jagannadh, Veerendra Kalyan; Murthy, Rashmi Sreeramachandra; Srinivasan, Rajesh; Gorthi, Sai Siva

    2016-06-01

    In this article, a portable microfluidic microscopy based approach for automated cytological investigations is presented. Inexpensive optical and electronic components have been used to construct a simple microfluidic microscopy system. In contrast to the conventional slide-based methods, the presented method employs microfluidics to enable automated sample handling and image acquisition. The approach involves the use of simple in-suspension staining and automated image acquisition to enable quantitative cytological analysis of samples. The applicability of the presented approach to research in cellular biology is shown by performing an automated cell viability assessment on a given population of yeast cells. Further, the relevance of the presented approach to clinical diagnosis and prognosis has been demonstrated by performing detection and differential assessment of malaria infection in a given sample. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Using Intensive Longitudinal Data Collected via Mobile Phone to Detect Imminent Lapse in Smokers Undergoing a Scheduled Quit Attempt.

    PubMed

    Businelle, Michael S; Ma, Ping; Kendzor, Darla E; Frank, Summer G; Wetter, David W; Vidrine, Damon J

    2016-10-17

    Mobile phone‒based real-time ecological momentary assessments (EMAs) have been used to record health risk behaviors, and antecedents to those behaviors, as they occur in near real time. The objective of this study was to determine if intensive longitudinal data, collected via mobile phone, could be used to identify imminent risk for smoking lapse among socioeconomically disadvantaged smokers seeking smoking cessation treatment. Participants were recruited into a randomized controlled smoking cessation trial at an urban safety-net hospital tobacco cessation clinic. All participants completed in-person EMAs on mobile phones provided by the study. The presence of six commonly cited lapse risk variables (ie, urge to smoke, stress, recent alcohol consumption, interaction with someone smoking, cessation motivation, and cigarette availability) collected during 2152 prompted or self-initiated postcessation EMAs was examined to determine whether the number of lapse risk factors was greater when lapse was imminent (ie, within 4 hours) than when lapse was not imminent. Various strategies were used to weight variables in efforts to improve the predictive utility of the lapse risk estimator. Participants (N=92) were mostly female (52/92, 57%), minority (65/92, 71%), 51.9 (SD 7.4) years old, and smoked 18.0 (SD 8.5) cigarettes per day. EMA data indicated significantly higher urges (P=.01), stress (P=.002), alcohol consumption (P<.001), interaction with someone smoking (P<.001), and lower cessation motivation (P=.03) within 4 hours of the first lapse compared with EMAs collected when lapse was not imminent. Further, the total number of lapse risk factors present within 4 hours of lapse (mean 2.43, SD 1.37) was significantly higher than the number of lapse risk factors present during periods when lapse was not imminent (mean 1.35, SD 1.04), P<.001. Overall, 62% (32/52) of all participants who lapsed completed at least one EMA wherein they reported ≥3 lapse risk factors within

  6. Imaging high stage river-water intrusion into a contaminated aquifer along a major river corridor using 2D time-lapse surface electrical resistivity tomography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wallin, Erin L.; Johnson, Timothy C.; Greenwood, William J.

    2013-03-29

    The Hanford 300 Area is located adjacent to the Columbia River in south-central Washington State, USA, and was a former site for nuclear fuel processing operations. Waste disposal practices resulted in persistent unsaturated zone and groundwater contamination, the primary contaminant of concern being uranium. Uranium behavior at the site is intimately linked with river stage driven groundwater-river water exchange such that understanding the nature of river water intrusion into the 300 Area is critical for predicting uranium desorption and transport. In this paper we use time-lapse electrical resistivity tomography (ERT) to image the inland intrusion of river during high stagemore » conditions. We demonstrate a modified time-lapse inversion approach, whereby the transient water table elevation is explicitly modeled by removing regularization constraints across the water table boundary. This implementation was critical for producing meaningful imaging results. We inverted approximately 1200 data sets (400 per line over 3 lines) using high performance computing resources to produce a time-lapse sequence of changes in bulk conductivity caused by river water intrusion during the 2011 spring runoff cycle over approximately 125 days. The resulting time series for each mesh element was then analyzed using common time series analysis to reveal the timing and location of river water intrusion beneath each line. The results reveal non-uniform flows characterized by preferred flow zones where river water enters and exits quickly with stage increase and decrease, and low permeability zones with broader bulk conductivity ‘break through’ curves and longer river water residence times. The time-lapse ERT inversion approach removes the deleterious effects of changing water table elevation and enables remote and spatial continuous groundwater-river water exchange monitoring using surface based ERT arrays under conditions where groundwater and river water conductivity are in

  7. Determinants of return behavior: a comparison of current and lapsed donors.

    PubMed

    Germain, Marc; Glynn, Simone A; Schreiber, George B; Gélinas, Stéphanie; King, Melissa; Jones, Mike; Bethel, James; Tu, Yongling

    2007-10-01

    There is a need to identify factors explaining why some people stop donating blood. A random mail survey of first-time (FT) and repeat (RPT) current (donating within 6 months before survey) and lapsed (donating >2 years prior) donors was conducted. The self-administered questionnaire included questions on personal, social, and behavioral characteristics. Among 1280 current and 1672 lapsed donors with valid addresses, the participation rate was 66.8 and 39.2 percent, respectively. In FT donors, the odds of lapsing increased with education (odds ratio [OR], 2.18; 95% confidence interval [CI], 1.34-3.55 for college or higher vs. Grade 12 or less education). Lapsed FT donors were more often asked to donate (OR, 1.89; 95% CI, 1.32-2.70) and had less interest in incentives (p < 0.001) than current FT donors. In RPT donors, lapsed status was associated with being younger (p < 0.001) and female (OR, 1.19; 95% CI, 1.00-1.42). Lapsed status was inversely associated with satisfaction with the last donation experience in both FT (p = 0.043) and RPT (p < 0.001) donors. Lapsed and current donors did not differ in perceived need for blood, personal transfusion experience, or mean reported altruistic behavior score. A positive donation experience appears to be a major determinant of donor return behavior. Lapsed donors do not appear, on average, to engage in fewer altruistic behaviors than currently active donors. Retention marketing strategies that appeal solely to altruistic values need to be further evaluated for their effectiveness.

  8. Simultaneous fluorescence and quantitative phase microscopy with single-pixel detectors

    NASA Astrophysics Data System (ADS)

    Liu, Yang; Suo, Jinli; Zhang, Yuanlong; Dai, Qionghai

    2018-02-01

    Multimodal microscopy offers high flexibilities for biomedical observation and diagnosis. Conventional multimodal approaches either use multiple cameras or a single camera spatially multiplexing different modes. The former needs expertise demanding alignment and the latter suffers from limited spatial resolution. Here, we report an alignment-free full-resolution simultaneous fluorescence and quantitative phase imaging approach using single-pixel detectors. By combining reference-free interferometry with single-pixel detection, we encode the phase and fluorescence of the sample in two detection arms at the same time. Then we employ structured illumination and the correlated measurements between the sample and the illuminations for reconstruction. The recovered fluorescence and phase images are inherently aligned thanks to single-pixel detection. To validate the proposed method, we built a proof-of-concept setup for first imaging the phase of etched glass with the depth of a few hundred nanometers and then imaging the fluorescence and phase of the quantum dot drop. This method holds great potential for multispectral fluorescence microscopy with additional single-pixel detectors or a spectrometer. Besides, this cost-efficient multimodal system might find broad applications in biomedical science and neuroscience.

  9. Gigavision - A weatherproof, multibillion pixel resolution time-lapse camera system for recording and tracking phenology in every plant in a landscape

    NASA Astrophysics Data System (ADS)

    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.

  10. Live Cells as Dynamic Laboratories: Time Lapse Raman Spectral Microscopy of Nanoparticles with Both IgE Targeting and pH-Sensing Functions

    DOE PAGES

    Nowak-Lovato, Kristy L.; Rector, Kirk D.

    2012-01-01

    Tmore » his review captures the use of live cells as dynamic microlaboratories through implementation of labeled nanoparticles (nanosensors) that have both sensing and targeting functions. he 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. o 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. his 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 + APase 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. he versatility of these probes shows promise for future applications relevant to intracellular trafficking and intelligent drug design.« less

  11. Time-lapse monitoring of TLR2 ligand internalization with newly developed fluorescent probes.

    PubMed

    Arai, Yohei; Yokoyama, Kouhei; Kawahara, Yuki; Feng, Qi; Ohta, Ippei; Shimoyama, Atsushi; Inuki, Shinsuke; Fukase, Koichi; Kabayama, Kazuya; Fujimoto, Yukari

    2018-05-23

    As a mammalian toll-like receptor family member protein, TLR2 recognizes lipoproteins from bacteria and modulates the immune response by inducing the expression of various cytokines. We have developed fluorescence-labeled TLR2 ligands with either hydrophilic or hydrophobic fluorescence groups. The labeled ligands maintained the inflammatory IL-6 induction activity and enabled us to observe the internalization and colocalization of the TLR2 ligands using live-cell imaging. The time-lapse monitoring in the live-cell imaging of the fluorescence-labeled TLR2 ligand showed that TLR2/CD14 expression in the host cells enhanced the internalization of TLR2 ligand molecules.

  12. Quantitative IR microscopy and spectromics open the way to 3D digital pathology.

    PubMed

    Bobroff, Vladimir; Chen, Hsiang-Hsin; Delugin, Maylis; Javerzat, Sophie; Petibois, Cyril

    2017-04-01

    Currently, only mass-spectrometry (MS) microscopy brings a quantitative analysis of chemical contents of tissue samples in 3D. Here, the reconstruction of a 3D quantitative chemical images of a biological tissue by FTIR spectro-microscopy is reported. An automated curve-fitting method is developed to extract all intense absorption bands constituting IR spectra. This innovation benefits from three critical features: (1) the correction of raw IR spectra to make them quantitatively comparable; (2) the automated and iterative data treatment allowing to transfer the IR-absorption spectrum into a IR-band spectrum; (3) the reconstruction of an 3D IR-band matrix (x, y, z for voxel position and a 4 th dimension with all IR-band parameters). Spectromics, which is a new method for exploiting spectral data for tissue metadata reconstruction, is proposed to further translate the related chemical information in 3D, as biochemical and anatomical tissue parameters. An example is given with oxidative stress distribution and the reconstruction of blood vessels in tissues. The requirements of IR microscopy instrumentation to propose 3D digital histology as a clinical routine technology is briefly discussed. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Quantitative comparison of 3D third harmonic generation and fluorescence microscopy images.

    PubMed

    Zhang, Zhiqing; Kuzmin, Nikolay V; Groot, Marie Louise; de Munck, Jan C

    2018-01-01

    Third harmonic generation (THG) microscopy is a label-free imaging technique that shows great potential for rapid pathology of brain tissue during brain tumor surgery. However, the interpretation of THG brain images should be quantitatively linked to images of more standard imaging techniques, which so far has been done qualitatively only. We establish here such a quantitative link between THG images of mouse brain tissue and all-nuclei-highlighted fluorescence images, acquired simultaneously from the same tissue area. For quantitative comparison of a substantial pair of images, we present here a segmentation workflow that is applicable for both THG and fluorescence images, with a precision of 91.3 % and 95.8 % achieved respectively. We find that the correspondence between the main features of the two imaging modalities amounts to 88.9 %, providing quantitative evidence of the interpretation of dark holes as brain cells. Moreover, 80 % bright objects in THG images overlap with nuclei highlighted in the fluorescence images, and they are 2 times smaller than the dark holes, showing that cells of different morphologies can be recognized in THG images. We expect that the described quantitative comparison is applicable to other types of brain tissue and with more specific staining experiments for cell type identification. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Time-Lapse Video Microscopy of Gliding Motility in Toxoplasma gondii Reveals a Novel, Biphasic Mechanism of Cell LocomotionV⃞

    PubMed Central

    Håkansson, Sebastian; Morisaki, Hiroshi; Heuser, John; Sibley, L. David

    1999-01-01

    Toxoplasma gondii is a member of the phylum Apicomplexa, a diverse group of intracellular parasites that share a unique form of gliding motility. Gliding is substrate dependent and occurs without apparent changes in cell shape and in the absence of traditional locomotory organelles. Here, we demonstrate that gliding is characterized by three distinct forms of motility: circular gliding, upright twirling, and helical rotation. Circular gliding commences while the crescent-shaped parasite lies on its right side, from where it moves in a counterclockwise manner at a rate of ∼1.5 μm/s. Twirling occurs when the parasite rights itself vertically, remaining attached to the substrate by its posterior end and spinning clockwise. Helical gliding is similar to twirling except that it occurs while the parasite is positioned horizontally, resulting in forward movement that follows the path of a corkscrew. The parasite begins lying on its left side (where the convex side is defined as dorsal) and initiates a clockwise revolution along the long axis of the crescent-shaped body. Time-lapse video analyses indicated that helical gliding is a biphasic process. During the first 180o of the turn, the parasite moves forward one body length at a rate of ∼1–3 μm/s. In the second phase, the parasite flips onto its left side, in the process undergoing little net forward motion. All three forms of motility were disrupted by inhibitors of actin filaments (cytochalasin D) and myosin ATPase (butanedione monoxime), indicating that they rely on an actinomyosin motor in the parasite. Gliding motility likely provides the force for active penetration of the host cell and may participate in dissemination within the host and thus is of both fundamental and practical interest. PMID:10564254

  15. A laboratory validation study of the time-lapse oscillatory pumping test for leakage detection in geological repositories

    NASA Astrophysics Data System (ADS)

    Sun, Alexander Y.; Lu, Jiemin; Islam, Akand

    2017-05-01

    Geologic repositories are extensively used for disposing byproducts in mineral and energy industries. The safety and reliability of these repositories are a primary concern to environmental regulators and the public. Time-lapse oscillatory pumping test (OPT) has been introduced recently as a pressure-based technique for detecting potential leakage in geologic repositories. By routinely conducting OPT at a number of pulsing frequencies, an operator may identify the potential repository anomalies in the frequency domain, alleviating the ambiguity caused by reservoir noise and improving the signal-to-noise ratio. Building on previous theoretical and field studies, this work performed a series of laboratory experiments to validate the concept of time-lapse OPT using a custom made, stainless steel tank under relatively high pressures. The experimental configuration simulates a miniature geologic storage repository consisting of three layers (i.e., injection zone, caprock, and above-zone aquifer). Results show that leakage in the injection zone led to deviations in the power spectrum of observed pressure data, and the amplitude of which also increases with decreasing pulsing frequencies. The experimental results are further analyzed by developing a 3D flow model, using which the model parameters are estimated through frequency domain inversion.

  16. A laboratory validation study of the time-lapse oscillatory pumping test concept for leakage detection in geological repositories

    NASA Astrophysics Data System (ADS)

    Sun, A. Y.; Islam, A.; Lu, J.

    2017-12-01

    Time-lapse oscillatory pumping test (OPT) has been introduced recently as a pressure-based monitoring technique for detecting potential leakage in geologic repositories. By routinely conducting OPT at a number of pulsing frequencies, a site operator may identify the potential anomalies in the frequency domain, alleviating the ambiguity caused by reservoir noise and improving the signal-to-noise ratio. Building on previous theoretical and field studies, this work performed a series of laboratory experiments to validate the concept of time-lapse OPT using a custom made, stainless steel tank under relatively high pressures ( 120psi). The experimental configuration simulates a miniature geologic storage repository consisting of three layers (i.e., injection zone, caprock, and above-zone aquifer). Results show that leakage in the injection zone led to deviations in the power spectrum of observed pressure data, and the amplitude of which also increases with decreasing pulsing frequencies. The experimental results were further analyzed by developing a 3D flow model, using which the model parameters were estimated through frequency domain inversion.

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

    NASA Astrophysics Data System (ADS)

    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.

  18. Things Forgotten: Simple Lapse or Serious Problem?

    MedlinePlus

    ... part of the U.S. Department of Health and Human Services Search form Search ... went there. And misplaced your keys or eyeglasses at least a few times. Many people worry about these memory lapses. They fear they’re heading toward a ...

  19. On the Resolvability of Steam Assisted Gravity Drainage Reservoirs Using Time-Lapse Gravity Gradiometry

    NASA Astrophysics Data System (ADS)

    Elliott, E. Judith; Braun, Alexander

    2017-11-01

    Unconventional heavy oil resource plays are important contributors to oil and gas production, as well as controversial for posing environmental hazards. Monitoring those reservoirs before, during, and after operations would assist both the optimization of economic benefits and the mitigation of potential environmental hazards. This study investigates how gravity gradiometry using superconducting gravimeters could resolve depletion areas in steam assisted gravity drainage (SAGD) reservoirs. This is achieved through modelling of a SAGD reservoir at 1.25 and 5 years of operation. Specifically, the density change structure identified from geological, petrological, and seismic observations is forward modelled for gravity and gradients. Three main parameters have an impact on the resolvability of bitumen depletion volumes and are varied through a suitable parameter space: well pair separation, depth to the well pairs, and survey grid sampling. The results include a resolvability matrix, which identifies reservoirs that could benefit from time-lapse gravity gradiometry monitoring. After 1.25 years of operation, during the rising phase, the resolvable maximum reservoir depth ranges between the surface and 230 m, considering a well pair separation between 80 and 200 m. After 5 years of production, during the spreading phase, the resolvability of depletion volumes around single well pairs is greatly compromised as the depletion volume is closer to the surface, which translates to a larger portion of the gravity signal. The modelled resolvability matrices were derived from visual inspection and spectral analysis of the gravity gradient signatures and can be used to assess the applicability of time-lapse gradiometry to monitor reservoir density changes.

  20. Live-cell confocal microscopy and quantitative 4D image analysis of anchor cell invasion through the basement membrane in C. elegans

    PubMed Central

    Kelley, Laura C.; Wang, Zheng; Hagedorn, Elliott J.; Wang, Lin; Shen, Wanqing; Lei, Shijun; Johnson, Sam A.; Sherwood, David R.

    2018-01-01

    Cell invasion through basement membrane (BM) barriers is crucial during development, leukocyte trafficking, and for the spread of cancer. Despite its importance in normal and diseased states, the mechanisms that direct invasion are poorly understood, in large part because of the inability to visualize dynamic cell-basement membrane interactions in vivo. This protocol describes multi-channel time-lapse confocal imaging of anchor cell invasion in live C. elegans. Methods presented include outline slide preparation and worm growth synchronization (15 min), mounting (20 min), image acquisition (20-180 min), image processing (20 min), and quantitative analysis (variable timing). Images acquired enable direct measurement of invasive dynamics including invadopodia formation, cell membrane protrusions, and BM removal. This protocol can be combined with genetic analysis, molecular activity probes, and optogenetic approaches to uncover molecular mechanisms underlying cell invasion. These methods can also be readily adapted for real-time analysis of cell migration, basement membrane turnover, and cell membrane dynamics by any worm laboratory. PMID:28880279

  1. Subsurface Hydrologic Processes Revealed by Time-lapse GPR in Two Contrasting Soils in the Shale Hills CZO

    NASA Astrophysics Data System (ADS)

    Guo, L.; Lin, H.; Nyquist, J.; Toran, L.; Mount, G.

    2017-12-01

    Linking subsurface structures to their functions in determining hydrologic processes, such as soil moisture dynamics, subsurface flow patterns, and discharge behaviours, is a key to understanding and modelling hydrological systems. Geophysical techniques provide a non-invasive approach to investigate this form-function dualism of subsurface hydrology at the field scale, because they are effective in visualizing subsurface structure and monitoring the distribution of water. In this study, we used time-lapse ground-penetrating radar (GPR) to compare the hydrologic responses of two contrasting soils in the Shale Hills Critical Zone Observatory. By integrating time-lapse GPR with artificial water injection, we observed distinct flow patterns in the two soils: 1) in the deep Rushtown soil (over 1.5 m depth to bedrock) located in a concave hillslope, a lateral preferential flow network extending as far as 2 m downslope was identified above a less permeable layer and via a series of connected macropores; whereas 2) in the shallow Weikert soil ( 0.3 m depth to saprock) located in a planar hillslope, vertical infiltration into the permeable fractured shale dominated the flow field, while the development of lateral preferential flow along the hillslope was restrained. At the Weikert soil site, the addition of brilliant blue dye to the water injection followed by in situ excavation supported GPR interpretation that only limited lateral preferential flow formed along the soil-saprock interface. Moreover, seasonally repeated GPR surveys indicated different patterns of profile moisture distribution in the two soils that in comparison with the dry season, a dense layer within the BC horizon in the deep Rushtown soil prevented vertical infiltration in the wet season, leading to the accumulation of soil moisture above this layer; whereas, in the shallow Weikert soil, water infiltrated into saprock in wet seasons, building up water storage within the fractured bedrock (i.e., the

  2. Quantitative segmentation of fluorescence microscopy images of heterogeneous tissue: Approach for tuning algorithm parameters

    NASA Astrophysics Data System (ADS)

    Mueller, Jenna L.; Harmany, Zachary T.; Mito, Jeffrey K.; Kennedy, Stephanie A.; Kim, Yongbaek; Dodd, Leslie; Geradts, Joseph; Kirsch, David G.; Willett, Rebecca M.; Brown, J. Quincy; Ramanujam, Nimmi

    2013-02-01

    The combination of fluorescent contrast agents with microscopy is a powerful technique to obtain real time images of tissue histology without the need for fixing, sectioning, and staining. The potential of this technology lies in the identification of robust methods for image segmentation and quantitation, particularly in heterogeneous tissues. Our solution is to apply sparse decomposition (SD) to monochrome images of fluorescently-stained microanatomy to segment and quantify distinct tissue types. The clinical utility of our approach is demonstrated by imaging excised margins in a cohort of mice after surgical resection of a sarcoma. Representative images of excised margins were used to optimize the formulation of SD and tune parameters associated with the algorithm. Our results demonstrate that SD is a robust solution that can advance vital fluorescence microscopy as a clinically significant technology.

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

    NASA Astrophysics Data System (ADS)

    Gosselet, A.; Singh, S. C.

    2007-12-01

    numerical simulation and can account for complex propagation effects as encountered in the Sleipner time-lapse data. This makes possible quantitative estimation of P and S-wave velocities on the meter scale. We applied the technique to 2D lines from the 1994, 1999 and 2006 vintages. The resulting post- injection models were subtracted to the pre-injection model to determine both the geometry and the velocity structure of the gas bearing areas which will be used to quantify the amount of CO2 in different forms (free versus dissolved).

  4. Predictors of smoking lapse in a human laboratory paradigm.

    PubMed

    Roche, Daniel J O; Bujarski, Spencer; Moallem, Nathasha R; Guzman, Iris; Shapiro, Jenessa R; Ray, Lara A

    2014-07-01

    During a smoking quit attempt, a single smoking lapse is highly predictive of future relapse. While several risk factors for a smoking lapse have been identified during clinical trials, a laboratory model of lapse was until recently unavailable and, therefore, it is unclear whether these characteristics also convey risk for lapse in a laboratory environment. The primary study goal was to examine whether real-world risk factors of lapse are also predictive of smoking behavior in a laboratory model of smoking lapse. After overnight abstinence, 77 smokers completed the McKee smoking lapse task, in which they were presented with the choice of smoking or delaying in exchange for monetary reinforcement. Primary outcome measures were the latency to initiate smoking behavior and the number of cigarettes smoked during the lapse. Several baseline measures of smoking behavior, mood, and individual traits were examined as predictive factors. Craving to relieve the discomfort of withdrawal, withdrawal severity, and tension level were negatively predictive of latency to smoke. In contrast, average number of cigarettes smoked per day, withdrawal severity, level of nicotine dependence, craving for the positive effects of smoking, and craving to relieve the discomfort of withdrawal were positively predictive of number of cigarettes smoked. The results suggest that real-world risk factors for smoking lapse are also predictive of smoking behavior in a laboratory model of lapse. Future studies using the McKee lapse task should account for between subject differences in the unique factors that independently predict each outcome measure.

  5. Integrating Terrestrial Time-Lapse Photography with Laser Scanning to Distinguish the Drivers of Movement at Sólheimajökull, Iceland

    NASA Astrophysics Data System (ADS)

    How, P.; James, M. R.; Wynn, P.

    2014-12-01

    Glacier movement is attributed to a sensitive configuration of driving forces. Here, we present an approach designed to evaluate the drivers of movement at Sólheimajökull, an outlet glacier from the Myrdalsjökull ice cap, Iceland, through combining terrestrial time-lapse photography and laser scanning (TLS). A time-lapse camera (a dSLR with intervalometer and solar-recharged battery power supply) collected hourly data over the summer of 2013. The data are subject to all the difficulties that are usually present in long time-lapse sequences, such as highly variable illumination and visibility conditions, evolving surfaces, and camera instabilities. Feature-tracking software [1] was used to: 1) track regions of static topography (e.g. the skyline) from which camera alignment could be continuously updated throughout the sequence; and 2) track glacial surface features for velocity estimation. Absolute georeferencing of the image sequence was carried out by registering the camera to a TLS survey acquired at the beginning of the monitoring period. A second TLS survey (July 2013) provided an additional 3D surface. By assuming glacial features moved in approximately planimetrically straight lines between the two survey dates, combining the two TLS surfaces with the monoscopic feature tracking allows 3D feature tracks to be derived. Such tracks will enable contributions from different drivers (e.g. surface melting) to be extracted, even in imagery that is acquired not perpendicular to glacier motion. At Sólheimajökull, our aim is to elucidate any volcanic contribution to the observed movement.[1] http://www.lancaster.ac.uk/staff/jamesm/software/pointcatcher.htm

  6. Active focus stabilization for upright selective plane illumination microscopy

    PubMed Central

    Hedde, Per Niklas; Gratton, Enrico

    2015-01-01

    Due to its sectioning capability, large field of view, and minimal light exposure, selective plane illumination microscopy has become the preferred choice for 3D time lapse imaging. Single cells in a dish can be conveniently imaged using an upright/inverted configuration. However, for measurements on long time scales (hours to days), mechanical drift is a problem; especially for studies of mammalian cells that typically require heating to 37°C which causes a thermal gradient across the instrument. Since the light sheet diverges towards the edges of the field of view, such a drift leads to a decrease in axial resolution over time. Or, even worse, the specimen could move out of the imaging volume. Here, we present a simple, cost-effective way to stabilize the axial position using the microscope camera to track the sample position. Thereby, sample loss is prevented and an optimal axial resolution is maintained by keeping the sample at the position where the light sheet is at its thinnest. We demonstrate the virtue of our approach by measurements of the light sheet thickness and 3D time lapse imaging of a cell monolayer at physiological conditions. PMID:26072829

  7. Active focus stabilization for upright selective plane illumination microscopy.

    PubMed

    Hedde, Per Niklas; Gratton, Enrico

    2015-06-01

    Due to its sectioning capability, large field of view, and minimal light exposure, selective plane illumination microscopy has become the preferred choice for 3D time lapse imaging. Single cells in a dish can be conveniently imaged using an upright/inverted configuration. However, for measurements on long time scales (hours to days), mechanical drift is a problem; especially for studies of mammalian cells that typically require heating to 37°C which causes a thermal gradient across the instrument. Since the light sheet diverges towards the edges of the field of view, such a drift leads to a decrease in axial resolution over time. Or, even worse, the specimen could move out of the imaging volume. Here, we present a simple, cost-effective way to stabilize the axial position using the microscope camera to track the sample position. Thereby, sample loss is prevented and an optimal axial resolution is maintained by keeping the sample at the position where the light sheet is at its thinnest. We demonstrate the virtue of our approach by measurements of the light sheet thickness and 3D time lapse imaging of a cell monolayer at physiological conditions.

  8. Flipping interferometry and its application for quantitative phase microscopy in a micro-channel.

    PubMed

    Roitshtain, Darina; Turko, Nir A; Javidi, Bahram; Shaked, Natan T

    2016-05-15

    We present a portable, off-axis interferometric module for quantitative phase microscopy of live cells, positioned at the exit port of a coherently illuminated inverted microscope. The module creates on the digital camera an interference pattern between the image of the sample and its flipped version. The proposed simplified module is based on a retro-reflector modification in an external Michelson interferometer. The module does not contain any lenses, pinholes, or gratings and its alignment is straightforward. Still, it allows full control of the off-axis angle and does not suffer from ghost images. As experimentally demonstrated, the module is useful for quantitative phase microscopy of live cells rapidly flowing in a micro-channel.

  9. The association of positive emotion and first smoking lapse: An ecological momentary assessment study.

    PubMed

    Vinci, Christine; Li, Liang; Wu, Cai; Lam, Cho Y; Guo, Lin; Correa-Fernández, Virmarie; Spears, Claire A; Hoover, Diana S; Etcheverry, Paul E; Wetter, David W

    2017-11-01

    Individuals attempting to quit smoking typically have poor success rates, and the majority fail to maintain long-term abstinence. Although a large body of evidence documents the impact of negative affect on reducing abstinence, there is a much smaller body of research on positive emotions, which could be an important mechanism that is associated with successful cessation. As such, this study examined positive emotions in real-time via ecological momentary assessment (EMA) to determine whether discrete positive emotions were uniquely related to 2 cessation milestones: quit day lapse and first lapse. Participants were 391 smokers who received tobacco cessation treatment. EMAs were completed pre- and postquit, and positive emotion was assessed with 3 items (enthusiastic, happy, and relaxed) rated on 5-point Likert scales. Analyses examined the associations of the means and slopes of each emotion on the current day with the likelihood of lapse on the following day. When controlling for relevant covariates, prequit positive emotions were not related to quit day lapse. However, postquit positive emotions were associated with first lapse. Specifically, high levels of happiness and relaxation, as well as increasing levels of enthusiasm, happiness, and relaxation were related to a lower likelihood of next day lapse. These are some of the first real-time, real-world data to demonstrate that distinct positive emotions are associated with a lower risk of lapse during the postquit period among smokers attempting to quit. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  10. Massively parallel data processing for quantitative total flow imaging with optical coherence microscopy and tomography

    NASA Astrophysics Data System (ADS)

    Sylwestrzak, Marcin; Szlag, Daniel; Marchand, Paul J.; Kumar, Ashwin S.; Lasser, Theo

    2017-08-01

    We present an application of massively parallel processing of quantitative flow measurements data acquired using spectral optical coherence microscopy (SOCM). The need for massive signal processing of these particular datasets has been a major hurdle for many applications based on SOCM. In view of this difficulty, we implemented and adapted quantitative total flow estimation algorithms on graphics processing units (GPU) and achieved a 150 fold reduction in processing time when compared to a former CPU implementation. As SOCM constitutes the microscopy counterpart to spectral optical coherence tomography (SOCT), the developed processing procedure can be applied to both imaging modalities. We present the developed DLL library integrated in MATLAB (with an example) and have included the source code for adaptations and future improvements. Catalogue identifier: AFBT_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AFBT_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: GNU GPLv3 No. of lines in distributed program, including test data, etc.: 913552 No. of bytes in distributed program, including test data, etc.: 270876249 Distribution format: tar.gz Programming language: CUDA/C, MATLAB. Computer: Intel x64 CPU, GPU supporting CUDA technology. Operating system: 64-bit Windows 7 Professional. Has the code been vectorized or parallelized?: Yes, CPU code has been vectorized in MATLAB, CUDA code has been parallelized. RAM: Dependent on users parameters, typically between several gigabytes and several tens of gigabytes Classification: 6.5, 18. Nature of problem: Speed up of data processing in optical coherence microscopy Solution method: Utilization of GPU for massively parallel data processing Additional comments: Compiled DLL library with source code and documentation, example of utilization (MATLAB script with raw data) Running time: 1,8 s for one B-scan (150 × faster in comparison to the CPU

  11. Assessment of ice-dam collapse by time-lapse photos at the Perito Moreno glacier, Argentina

    NASA Astrophysics Data System (ADS)

    Lenzano, M. G.; Lannutti, E.; Toth, C. K.; Lenzano, L. E.; Lovecchio, A.

    2014-11-01

    This research provides a feasibility study on the implementation and performance assessment of time-lapse processing of a monoscopic image sequence, acquired by a calibrated camera in the Perito Moreno Glacier in Argentina. The glacier is located at 50°28'23" S, 73°02'10" W at the Parque Nacional Los Glaciares, South Patagonia Icefield, Santa Cruz and has experienced minor fluctuations and unusual behavior since the early 1960's to present. The objective of this study was to determine the evolution and changes in the ice-dam of the Perito Moreno glacier that started on November, 23 2012 and collapsed on January 19, 2013. Two images every 24 hours were acquired since October 2012 until February 2013, a total of 135 days. Image information was supported by ground data. Image and ground data was correlated with a 2D affine transformation. This technique allows the determination of the distortions in the images and estimating the values of scale factors. This, along with an accurate time-lapse interval, has produced accurate data for the analysis. In addition, changes in the level of the Brazo Rico lake were validated with direct data in order to determine the degree of uncertainty in the estimation of changes in the glacier. Based on the calculations, advance rates of the front of the Perito Moreno glacier were estimated at 0.67 m/d ± 0.003 m, and the tunnel evolution was also recorded.

  12. Time-lapse cameras as an aid in studying grizzly bears in northwest Wyoming

    USGS Publications Warehouse

    Ball, Ronald E.

    1980-01-01

    Time-lapse cameras were effective for gathering limited distribution and population data on grizzly bears (Ursus arctos) and black bears (Ursus americanus) in northwest Wyoming. Thirty-six stations, each consisting of a camera and a lure, were monitored for 551 camera-days; 83 rolls of film were exposed. Five different lures were tested. Thirty-one bears (5 grizzly, 25 black, 1 unknown bear) were identified at 15 stations. Young:adult and young:female ratios observed (0.4 and 1.5 for black bears and 0.7 and 2.0 for grizzlies) corresponded well with those of other researchers in the region. One sighting recorded on film extended the known range of the grizzly bear in the Shoshone National Forest.

  13. Novel aspects of live intestinal epithelial cell function revealed using a custom time-lapse video microscopy apparatus.

    PubMed

    Papetti, Michael; Kozlowski, Piotr

    2018-04-01

    Many aspects of cell physiology, including migration, membrane function, and cell division, are best understood by observing live cell dynamics over time using video microscopy. To probe these phenomena in colon epithelial cells using simple components with a limited budget, we have constructed an inexpensive (<$410) self-contained apparatus, consisting of a closed-loop, feedback-controlled system regulated by a PID (proportional-integrative-derivative) controller contained within a 0.077 m 3 insulated acrylic box. Temperature, humidity, pH, and proliferative capacity of colon epithelial cells in this system mimic those in a standard tissue culture incubator for over four days. Our system offers significant advantages over existing cost-prohibitive commercially available and custom-made devices because of its very low cost, use of PID temperature control, lack of reliance on constant infusion of external humidified, heated air or carbon dioxide, ability to directly measure cell culture medium temperature, and combination of exquisite cellular detail with minimal focus drift under physiological conditions for extended periods of time. Using this apparatus, coupled with an inverted microscope equipped with phase contrast optics and a programmable digital camera, we have observed many events in colon epithelial cells not visible by static imaging, including kinetics of normal and abnormal mitoses, dynamic membrane structures, intracellular vesicle movements, and cell migration. © 2018 International Society for Advancement of Cytometry. © 2018 International Society for Advancement of Cytometry.

  14. Single embryo transfer by Day 3 time-lapse selection versus Day 5 conventional morphological selection: a randomized, open-label, non-inferiority trial.

    PubMed

    Yang, Lanlin; Cai, Sufen; Zhang, Shuoping; Kong, Xiangyi; Gu, Yifan; Lu, Changfu; Dai, Jing; Gong, Fei; Lu, Guangxiu; Lin, Ge

    2018-05-01

    Does single cleavage-stage (Day 3) embryo transfer using a time-lapse (TL) hierarchical classification model achieve comparable ongoing pregnancy rates (OPR) to single blastocyst (Day 5) transfer by conventional morphological (CM) selection? Day 3 single embryo transfer (SET) with a hierarchical classification model had a significantly lower OPR compared with Day 5 SET with CM selection. Cleavage-stage SET is an alternative to blastocyst SET. Time-lapse imaging assists better embryo selection, based on studies of pregnancy outcomes when adding time-lapse imaging to CM selection at the cleavage or blastocyst stage. This single-centre, randomized, open-label, active-controlled, non-inferiority study included 600 women between October 2015 and April 2017. Eligible patients were Chinese females, aged ≤36 years, who were undergoing their first or second fresh IVF cycle using their own oocytes, and who had FSH levels ≤12 IU/mL on Day 3 of the cycle and 10 or more oocytes retrieved. Patients who had underlying uterine conditions, oocyte donation, recurrent pregnancy loss, abnormal oocytes or <6 normally fertilized embryos (2PN) were excluded from the study participation. Patients were randomized 1:1 to either the cleavage-stage SET with a time-lapse hierarchical classification model for selection (D3 + TL) or blastocyst SET with CM selection (D5 + CM). All normally fertilized zygotes were cultured in Primo Vision. The study was conducted at a tertiary IVF centre (CITIC-Xiangya) and OPR was the primary outcome. A total of 600 patients were randomized to the two groups, among which 585 (D3 + TL = 290, D5 + CM = 295) were included in the Modified-intention-to-treat (mITT) population and 517 (D3 + TL = 261, D5 + CM = 256) were included in the PP population. In the per protocol (PP) population, OPR was significantly lower in the D3 group (59.4%, 155/261) than in the D5 group (68.4%, 175/256) (difference: -9.0%, 95% CI: -17.1%, -0.7%, P = 0.03). Analysis in mITT population

  15. Momentary factors during marijuana use as predictors of lapse during attempted abstinence in young adults.

    PubMed

    Shrier, Lydia A; Sarda, Vishnudas; Jonestrask, Cassandra; Harris, Sion Kim

    2018-08-01

    Young adults using marijuana heavily often try multiple times to quit on their own. We sought to identify momentary experiences during marijuana use that could aid in predicting lapse when young adults subsequently attempt abstinence. Young adults (N=34) age 18-25 using marijuana ≥5days/week and planning to quit completed a survey of sociodemographic characteristics, substance use, marijuana expectancies, use motives, perceived social support, and confidence to abstain. They completed ecological momentary assessment (EMA) smartphone reports several times/day for two weeks prior to, then during two weeks of attempted abstinence. Use period EMA reports assessed affect, craving, accessibility, situational permissibility, use, and motivation to abstain. Baseline survey and EMA data were examined in relation to subsequent lapse during attempted abstinence. Nearly 3 in 4 participants (73.5%) reported lapsing during attempted abstinence from marijuana. On bivariate analyses, lower baseline dependence severity score, negative effect expectancies, perceived family support, and confidence to abstain were each associated with lapse. Of the use period EMA variables, greater percent of days with marijuana use, reports of easy accessibility, and reports of situational permissibility were each associated with lapse. Modeled together, negative effect expectancies, perceived family support, confidence to abstain, and situational permissibility during use were highly accurate in predicting lapse during attempted abstinence. Momentary factors may add to conventionally-surveyed characteristics to enhance prediction of lapse during attempted abstinence among young adults with heavy marijuana use. Momentary assessment prior to a quit attempt may thus enable more effective personalized approaches to preventing lapse. Copyright © 2018 Elsevier Ltd. All rights reserved.

  16. Zebrafish Caudal Fin Angiogenesis Assay—Advanced Quantitative Assessment Including 3-Way Correlative Microscopy

    PubMed Central

    Correa Shokiche, Carlos; Schaad, Laura; Triet, Ramona; Jazwinska, Anna; Tschanz, Stefan A.; Djonov, Valentin

    2016-01-01

    Background Researchers evaluating angiomodulating compounds as a part of scientific projects or pre-clinical studies are often confronted with limitations of applied animal models. The rough and insufficient early-stage compound assessment without reliable quantification of the vascular response counts, at least partially, to the low transition rate to clinics. Objective To establish an advanced, rapid and cost-effective angiogenesis assay for the precise and sensitive assessment of angiomodulating compounds using zebrafish caudal fin regeneration. It should provide information regarding the angiogenic mechanisms involved and should include qualitative and quantitative data of drug effects in a non-biased and time-efficient way. Approach & Results Basic vascular parameters (total regenerated area, vascular projection area, contour length, vessel area density) were extracted from in vivo fluorescence microscopy images using a stereological approach. Skeletonization of the vasculature by our custom-made software Skelios provided additional parameters including “graph energy” and “distance to farthest node”. The latter gave important insights into the complexity, connectivity and maturation status of the regenerating vascular network. The employment of a reference point (vascular parameters prior amputation) is unique for the model and crucial for a proper assessment. Additionally, the assay provides exceptional possibilities for correlative microscopy by combining in vivo-imaging and morphological investigation of the area of interest. The 3-way correlative microscopy links the dynamic changes in vivo with their structural substrate at the subcellular level. Conclusions The improved zebrafish fin regeneration model with advanced quantitative analysis and optional 3-way correlative morphology is a promising in vivo angiogenesis assay, well-suitable for basic research and preclinical investigations. PMID:26950851

  17. Motion estimation of subcellular structures from fluorescence microscopy images.

    PubMed

    Vallmitjana, A; Civera-Tregon, A; Hoenicka, J; Palau, F; Benitez, R

    2017-07-01

    We present an automatic image processing framework to study moving intracellular structures from live cell fluorescence microscopy. The system includes the identification of static and dynamic structures from time-lapse images using data clustering as well as the identification of the trajectory of moving objects with a probabilistic tracking algorithm. The method has been successfully applied to study mitochondrial movement in neurons. The approach provides excellent performance under different experimental conditions and is robust to common sources of noise including experimental, molecular and biological fluctuations.

  18. Quantitative Image Restoration in Bright Field Optical Microscopy.

    PubMed

    Gutiérrez-Medina, Braulio; Sánchez Miranda, Manuel de Jesús

    2017-11-07

    Bright field (BF) optical microscopy is regarded as a poor method to observe unstained biological samples due to intrinsic low image contrast. We introduce quantitative image restoration in bright field (QRBF), a digital image processing method that restores out-of-focus BF images of unstained cells. Our procedure is based on deconvolution, using a point spread function modeled from theory. By comparing with reference images of bacteria observed in fluorescence, we show that QRBF faithfully recovers shape and enables quantify size of individual cells, even from a single input image. We applied QRBF in a high-throughput image cytometer to assess shape changes in Escherichia coli during hyperosmotic shock, finding size heterogeneity. We demonstrate that QRBF is also applicable to eukaryotic cells (yeast). Altogether, digital restoration emerges as a straightforward alternative to methods designed to generate contrast in BF imaging for quantitative analysis. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

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

    NASA Astrophysics Data System (ADS)

    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.

  20. Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform

    NASA Astrophysics Data System (ADS)

    Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

    2015-11-01

    Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (r

  1. Monitoring channel head erosion processes in response to an artificially induced abrupt base level change using time-lapse photography 2301

    USDA-ARS?s Scientific Manuscript database

    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...

  2. Noninvasive quantitative measurement of colloid transport in mesoscale porous media using time lapse fluorescence imaging.

    PubMed

    Bridge, Jonathan W; Banwart, Steven A; Heathwaite, A Louise

    2006-10-01

    We demonstrate noninvasive quantitative imaging of colloid and solute transport at millimeter to decimeter (meso-) scale. Ultraviolet (UV) excited fluorescent solute and colloid tracers were independently measured simultaneously during co-advection through saturated quartz sand. Pulse-input experiments were conducted at constant flow rates and ionic strengths 10(-3), 10(-2) and 10(-1) M NaCl. Tracers were 1.9 microm carboxylate latex microspheres and disodium fluorescein. Spatial moments analysis was used to quantify relative changes in mass distribution of the colloid and solute tracers over time. The solute advected through the sand at a constant velocity proportional to flow rate and was described well by a conservative transport model (CXTFIT). In unfavorable deposition conditions increasing ionic strength produced significant reduction in colloid center of mass transport velocity over time. Velocity trends correlated with the increasing fraction of colloid mass retained along the flowpath. Attachment efficiencies (defined by colloid filtration theory) calculated from nondestructive retained mass data were 0.013 +/- 0.03, 0.09 +/- 0.02, and 0.22 +/- 0.05 at 10(-3), 10(-2), and 10(-1) M ionic strength, respectively, which compared well with previously published data from breakthrough curves and destructive sampling. Mesoscale imaging of colloid mass dynamics can quantify key deposition and transport parameters based on noninvasive, nondestructive, spatially high-resolution data.

  3. Optofluidic time-stretch microscopy: recent advances

    NASA Astrophysics Data System (ADS)

    Lei, Cheng; Nitta, Nao; Ozeki, Yasuyuki; Goda, Keisuke

    2018-06-01

    Flow cytometry is an indispensable method for valuable applications in numerous fields such as immunology, pathology, pharmacology, molecular biology, and marine biology. Optofluidic time-stretch microscopy is superior to conventional flow cytometry methods for its capability to acquire high-quality images of single cells at a high-throughput exceeding 10,000 cells per second. This makes it possible to extract copious information from cellular images for accurate cell detection and analysis with the assistance of machine learning. Optofluidic time-stretch microscopy has proven its effectivity in various applications, including microalga-based biofuel production, evaluation of thrombotic disorders, as well as drug screening and discovery. In this review, we discuss the principles and recent advances of optofluidic time-stretch microscopy.

  4. Optofluidic time-stretch microscopy: recent advances

    NASA Astrophysics Data System (ADS)

    Lei, Cheng; Nitta, Nao; Ozeki, Yasuyuki; Goda, Keisuke

    2018-04-01

    Flow cytometry is an indispensable method for valuable applications in numerous fields such as immunology, pathology, pharmacology, molecular biology, and marine biology. Optofluidic time-stretch microscopy is superior to conventional flow cytometry methods for its capability to acquire high-quality images of single cells at a high-throughput exceeding 10,000 cells per second. This makes it possible to extract copious information from cellular images for accurate cell detection and analysis with the assistance of machine learning. Optofluidic time-stretch microscopy has proven its effectivity in various applications, including microalga-based biofuel production, evaluation of thrombotic disorders, as well as drug screening and discovery. In this review, we discuss the principles and recent advances of optofluidic time-stretch microscopy.

  5. Time-Lapse Video of SLS Engine Section Test Article Being Stacked at Michoud

    NASA Image and Video Library

    2017-04-25

    This time-lapse video shows the Space Launch System engine section structural qualification test article being stacked at NASA's Michoud Assembly Facility in New Orleans. The rocket's engine section is the bottom of the core stage and houses the four RS-25 engines. The engine section test article was moved to Michoud's Cell A in Building 110 for vertical stacking with hardware that simulates the rocket's liquid hydrogen tank, which is the fuel tank that joins to the engine section. Once stacked, the entire test article will load onto the barge Pegasus and ship to NASA's Marshall Space Flight Center in Huntsville, Alabama. There, it will be subjected to millions of pounds of force during testing to ensure the hardware can withstand the incredible stresses of launch.

  6. Advances in Light Microscopy for Neuroscience

    PubMed Central

    Wilt, Brian A.; Burns, Laurie D.; Ho, Eric Tatt Wei; Ghosh, Kunal K.; Mukamel, Eran A.

    2010-01-01

    Since the work of Golgi and Cajal, light microscopy has remained a key tool for neuroscientists to observe cellular properties. Ongoing advances have enabled new experimental capabilities using light to inspect the nervous system across multiple spatial scales, including ultrastructural scales finer than the optical diffraction limit. Other progress permits functional imaging at faster speeds, at greater depths in brain tissue, and over larger tissue volumes than previously possible. Portable, miniaturized fluorescence microscopes now allow brain imaging in freely behaving mice. Complementary progress on animal preparations has enabled imaging in head-restrained behaving animals, as well as time-lapse microscopy studies in the brains of live subjects. Mouse genetic approaches permit mosaic and inducible fluorescence-labeling strategies, whereas intrinsic contrast mechanisms allow in vivo imaging of animals and humans without use of exogenous markers. This review surveys such advances and highlights emerging capabilities of particular interest to neuroscientists. PMID:19555292

  7. The influence of lapses of attention on working memory capacity.

    PubMed

    Unsworth, Nash; Robison, Matthew K

    2016-02-01

    In three experiments, the influence of lapses of attention on working memory (WM) capacity measures was examined. Participants performed various change detection tasks while also reporting whether they were focused on the current task or whether they were unfocused and mind-wandering. Participants reported that they were mind-wandering roughly 27% of the time, and when participants reported mind-wandering, their performance was worse compared to when they reported being on-task. Low WM capacity individuals reported more mind-wandering and lapses of attention than high WM capacity individuals, and mind-wandering and filtering abilities were shown to make independent contributions to capacity estimates. These results provide direct support for the notion that the ability to focus attention on-task and prevent lapses of attention is an important contributor to performance on measures of WM capacity.

  8. Utilizing time-lapse micro-CT-correlated bisphosphonate binding kinetics and soft tissue-derived input functions to differentiate site-specific changes in bone metabolism in vivo.

    PubMed

    Tower, R J; Campbell, G M; Müller, M; Glüer, C C; Tiwari, S

    2015-05-01

    The turnover of bone is a tightly regulated process between bone formation and resorption to ensure skeletal homeostasis. This process differs between bone types, with trabecular bone often associated with higher turnover than cortical bone. Analyses of bone by micro-computed tomography (micro-CT) reveal changes in structure and mineral content, but are limited in the study of metabolic activity at a single time point, while analyses of serum markers can reveal changes in bone metabolism, but cannot delineate the origin of any aberrant findings. To obtain a site-specific assessment of bone metabolic status, bisphosphonate binding kinetics were utilized. Using a fluorescently-labeled bisphosphonate, we show that early binding kinetics monitored in vivo using fluorescent molecular tomography (FMT) can monitor changes in bone metabolism in response to bone loss, stimulated by ovariectomy (OVX), or bone gain, resulting from treatment with the anabolic bone agent parathyroid hormone (PTH), and is capable of distinguishing different, metabolically distinct skeletal sites. Using time-lapse micro-CT, longitudinal bone turnover was quantified. The spine showed a significantly greater percent resorbing volume and surface in response to OVX, while mice treated with PTH showed significantly greater resorbing volume per bone surface in the spine and significantly greater forming surfaces in the knee. Correlation studies between binding kinetics and micro-CT suggest that forming surfaces, as assessed by time-lapse micro-CT, are preferentially reflected in the rate constant values while forming and resorbing bone volumes primarily affect plateau values. Additionally, we developed a blood pool correction method which now allows for quantitative multi-compartment analyses to be conducted using FMT. These results further expand our understanding of bisphosphonate binding and the use of bisphosphonate binding kinetics as a tool to monitor site-specific changes in bone metabolism in

  9. Monitoring water flows with time-lapse Electrical Resistivity Tomography on the Super-Sauze landslide

    NASA Astrophysics Data System (ADS)

    Gance, J.; Sailhac, P.; Malet, J.-P.; Grandjean, G.; Supper, R.; Jochum, B.; Ottowitz, D.

    2012-04-01

    This work presents results of a permanent hydro-geophysical monitoring of an active landslide developed in clay-shales. Hydrology has been proved to be a major factor controlling the Super-Sauze earthflow behavior, but it knowledge still limited mainly because of the importance of spatial heterogeneities. The geometry of the bedrock creates internal crests and gullies that can guide waterflows or create a lock and engender an excess of pore water pressure; the soil surface characteristics plays also a large role in the surface hydraulic conductivity, and therefore, on the infiltration pattern. To understand in detail these processes, it is therefore important to monitor spatially at large scale (with high resolution) those phenomena and to overcome the monitoring difficulties inherent to a fast-moving clayey earthflow. The objectives of the survey are to identify and characterize spatially and temporarily the water flow circulation within the landslide body over a period of one year. The studied profile measures 114 m long and is surveyed with 93 electrodes spaced from 0.5, 1 or 2 meter according the soil surface cracking. Four resistivity datasets of 4300 measurements are acquired each day using a gradient array since May 2011. The monitoring is performed with the GEOMON4D system, developed by the Geological Survey of Austria. To facilitate the interpretation, humidity, conductivity, temperature, and piezometer sensors are placed along the profile. Two dGPS antenna placed upstream and downstream the profile allow to correlate the results with soil displacement. Lefranc tests and granulometry results realized on several samples have shown the important heterogeneities of the near surface. The objective of this work is to present the data processing strategy for the analysis of long periods time-lapse ERT survey of natural rain events taking into account changes through time of the position of the electrodes, changes in the soil surface state and important changes

  10. Quantitative single-molecule imaging by confocal laser scanning microscopy.

    PubMed

    Vukojevic, Vladana; Heidkamp, Marcus; Ming, Yu; Johansson, Björn; Terenius, Lars; Rigler, Rudolf

    2008-11-25

    A new approach to quantitative single-molecule imaging by confocal laser scanning microscopy (CLSM) is presented. It relies on fluorescence intensity distribution to analyze the molecular occurrence statistics captured by digital imaging and enables direct determination of the number of fluorescent molecules and their diffusion rates without resorting to temporal or spatial autocorrelation analyses. Digital images of fluorescent molecules were recorded by using fast scanning and avalanche photodiode detectors. In this way the signal-to-background ratio was significantly improved, enabling direct quantitative imaging by CLSM. The potential of the proposed approach is demonstrated by using standard solutions of fluorescent dyes, fluorescently labeled DNA molecules, quantum dots, and the Enhanced Green Fluorescent Protein in solution and in live cells. The method was verified by using fluorescence correlation spectroscopy. The relevance for biological applications, in particular, for live cell imaging, is discussed.

  11. Stimulant Treatment Reduces Lapses in Attention among Children with ADHD: The Effects of Methylphenidate on Intra-Individual Response Time Distributions

    ERIC Educational Resources Information Center

    Spencer, Sarah V.; Hawk, Larry W., Jr.; Richards, Jerry B.; Shiels, Keri; Pelham, William E., Jr.; Waxmonsky, James G.

    2009-01-01

    Recent research has suggested that intra-individual variability in reaction time (RT) distributions of children with ADHD is characterized by a particularly large rightward skew that may reflect lapses in attention. The purpose of the study was to provide the first randomized, placebo-controlled test of the effects of the stimulant methylphenidate…

  12. Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

    PubMed

    Shrivastava, Sajal; Sohn, Il-Yung; Son, Young-Min; Lee, Won-Il; Lee, Nae-Eung

    2015-12-14

    Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

  13. PREFACE: Time-resolved scanning tunnelling microscopy Time-resolved scanning tunnelling microscopy

    NASA Astrophysics Data System (ADS)

    Zandvliet, Harold J. W.; Lin, Nian

    2010-07-01

    Scanning tunnelling microscopy has revolutionized our ability to image, manipulate, and investigate solid surfaces on the length scale of individual atoms and molecules. The strength of this technique lies in its imaging capabilities, since for many scientists 'seeing is believing'. However, scanning tunnelling microscopy also suffers from a severe limitation, namely its poor time resolution. Recording a scanning tunnelling microscopy image typically requires a few tens of seconds for a conventional scanning tunnelling microscope to a fraction of a second for a specially designed fast scanning tunnelling microscope. Designing and building such a fast scanning tunnelling microscope is a formidable task in itself and therefore, only a limited number of these microscopes have been built [1]. There is, however, another alternative route to significantly enhance the time resolution of a scanning tunnelling microscope. In this alternative method, the tunnelling current is measured as a function of time with the feedback loop switched off. The time resolution is determined by the bandwidth of the IV converter rather than the cut-off frequency of the feedback electronics. Such an approach requires a stable microscope and goes, of course, at the expense of spatial information. In this issue, we have collected a set of papers that gives an impression of the current status of this rapidly emerging field [2]. One of the very first attempts to extract information from tunnel current fluctuations was reported by Tringides' group in the mid-1990s [3]. They showed that the collective diffusion coefficient can be extracted from the autocorrelation of the time-dependent tunnelling current fluctuations produced by atom motion in and out of the tunnelling junction. In general, current-time traces provide direct information on switching/conformation rates and distributions of residence times. In the case where these processes are thermally induced it is rather straightforward to map

  14. Predicting the Initial Lapse Using a Mobile Health Application after Alcohol Detoxification

    ERIC Educational Resources Information Center

    Chih, Ming-Yuan

    2013-01-01

    The prediction and prevention of the initial lapse--which is defined as the first lapse after a period of abstinence--is important because the initial lapse often leads to subsequent lapses (within the same lapse episode) or relapse. The prediction of the initial lapse may allow preemptive intervention to be possible. This dissertation reports on…

  15. An automated real-time microscopy system for analysis of fluorescence resonance energy transfer

    NASA Astrophysics Data System (ADS)

    Bernardini, André; Wotzlaw, Christoph; Lipinski, Hans-Gerd; Fandrey, Joachim

    2010-05-01

    Molecular imaging based on Fluorescence Resonance Energy Transfer (FRET) is widely used in cellular physiology both for protein-protein interaction analysis and detecting conformational changes of single proteins, e.g. during activation of signaling cascades. However, getting reliable results from FRET measurements is still hampered by methodological problems such as spectral bleed through, chromatic aberration, focal plane shifts and false positive FRET. Particularly false positive FRET signals caused by random interaction of the fluorescent dyes can easily lead to misinterpretation of the data. This work introduces a Nipkow Disc based FRET microscopy system, that is easy to operate without expert knowledge of FRET. The system automatically accounts for all relevant sources of errors and provides various result presentations of two, three and four dimensional FRET data. Two examples are given to demonstrate the scope of application. An interaction analysis of the two subunits of the hypoxia-inducible transcription factor 1 demonstrates the use of the system as a tool for protein-protein interaction analysis. As an example for time lapse observations, the conformational change of the fluorophore labeled heat shock protein 33 in the presence of oxidant stress is shown.

  16. Quantitative comparison between full-spectrum and filter-based imaging in hyperspectral fluorescence microscopy

    PubMed Central

    GAO, L.; HAGEN, N.; TKACZYK, T.S.

    2012-01-01

    Summary We implement a filterless illumination scheme on a hyperspectral fluorescence microscope to achieve full-range spectral imaging. The microscope employs polarisation filtering, spatial filtering and spectral unmixing filtering to replace the role of traditional filters. Quantitative comparisons between full-spectrum and filter-based microscopy are provided in the context of signal dynamic range and accuracy of measured fluorophores’ emission spectra. To show potential applications, a five-colour cell immunofluorescence imaging experiment is theoretically simulated. Simulation results indicate that the use of proposed full-spectrum imaging technique may result in three times improvement in signal dynamic range compared to that can be achieved in the filter-based imaging. PMID:22356127

  17. 3D time-lapse analysis of Rab11/FIP5 complex: spatiotemporal dynamics during apical lumen formation.

    PubMed

    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.

  18. Near-surface air temperature lapse rates in Xinjiang, northwestern China

    NASA Astrophysics Data System (ADS)

    Du, Mingxia; Zhang, Mingjun; Wang, Shengjie; Zhu, Xiaofan; Che, Yanjun

    2018-02-01

    Lapse rates of near-surface (2 m) air temperature are important parameters in hydrologic and climate simulations, especially for the mountainous areas without enough in-situ observations. In Xinjiang, northwestern China, the elevations range from higher than 7000 m to lower than sea level, but the existing long-term meteorological measurements are limited and distributed unevenly. To calculate lapse rates in Xinjiang, the daily data of near-surface air temperature ( T min, T ave, and T max) were measured by automatic weather stations from 2012 to 2014. All the in situ observation stations were gridded into a network of 1.5° (latitude) by 1.5° (longitude), and the spatial distribution and the daily, monthly, seasonal variations of lapse rates for T min, T ave, and T max in Xinjiang are analyzed. The Urumqi River Basin has been considered as a case to study the influence of elevation, aspect, and the wet and dry air conditions to the T min, T ave, and T max lapse rates. Results show that (1) the lapse rates for T min, T ave, and T max vary spatially during the observation period. The spatial diversity of T min lapse rates is larger than that of T ave, and that of T max is the smallest. For each season, T max lapse rates have more negative values than T ave lapse rates which are steeper than T min lapse rates. The weakest spatial diversity usually appears in July throughout a year. (2) The comparison for the three subregions (North, Middle, and South region) exhibits that lapse rates have similar day-to-day and month-to-month characteristics which present shallower values in winter months and steeper values in summer months. The T ave lapse rates in North region are shallower than those in Middle and South region, and the steepest T ave lapse rates of the three regions all appear in April. T min lapse rates are shallower than T max lapse rates. The maximum medians of T min and T max lapse rates for each grid in the three regions all appear in January, whereas the

  19. Presentation of a High Resolution Time Lapse 3D Groundwater Model of Metsähovi for Calculating the Gravity Effect of Groundwater in Local Scale

    NASA Astrophysics Data System (ADS)

    Hokkanen, T. M.; Hartikainen, A.; Raja-Halli, A.; Virtanen, H.; Makinen, J.

    2015-12-01

    INTRODUCTION The aim of this study is to construct a fine resolution time lapse groundwater (GW) model of Metsähovi (MH). GW, geological, and soil moisture (SM) data were collected for several years to achieve the goal. The knowledge of the behavior of the GW at local scale is essential for superconductive gravimeter (SG) investigations performing in MH. DESCRIPTION OF THE DATA Almost 50 sensors have been recorded SM data some 6 years with 1 to 5 minutes sampling frequency. The GW table has been monitored, both in bedrock and in soil, in many stages with all together 15 piezometers. Two geological sampling campaigns were conducted to get the knowledge of hydrological properties of soil in the study area of 200×200 m2 around SG station in MH. PRINCIPLE OF TIME LAPSE 3D HYDROGEOLOGICAL MODEL The model of study site consists of the surfaces of ground and bedrock gridded with 2×2 m2 resolution. The height of GW table was interpolated to 2×2×0.1 m3 grid between GW and SM monitoring points. Close to the outline of the study site and areas lacking of sensors GW table was defined by extrapolation and considering the geological information of the area. The bedrock porosity is 2% and soil porosity determined by geological information and SM recordings is from 5 to 35%. Only fully saturated media is considered in the time lapse model excluding unsaturated one. BENEFICIERS With a new model the fluctuation of GW table can be followed with ranging time lapses from 1 minute to 1 month. The gravity effect caused by the variation of GW table can be calculated more accurate than before in MH. Moreover, the new model can be validated and refined by measured gravity, i.e. hydrological model can be improved by SG recordings (Figure 1).

  20. Weak-beam scanning transmission electron microscopy for quantitative dislocation density measurement in steels.

    PubMed

    Yoshida, Kenta; Shimodaira, Masaki; Toyama, Takeshi; Shimizu, Yasuo; Inoue, Koji; Yoshiie, Toshimasa; Milan, Konstantinovic J; Gerard, Robert; Nagai, Yasuyoshi

    2017-04-01

    To evaluate dislocations induced by neutron irradiation, we developed a weak-beam scanning transmission electron microscopy (WB-STEM) system by installing a novel beam selector, an annular detector, a high-speed CCD camera and an imaging filter in the camera chamber of a spherical aberration-corrected transmission electron microscope. The capabilities of the WB-STEM with respect to wide-view imaging, real-time diffraction monitoring and multi-contrast imaging are demonstrated using typical reactor pressure vessel steel that had been used in an European nuclear reactor for 30 years as a surveillance test piece with a fluence of 1.09 × 1020 neutrons cm-2. The quantitatively measured size distribution (average loop size = 3.6 ± 2.1 nm), number density of the dislocation loops (3.6 × 1022 m-3) and dislocation density (7.8 × 1013 m m-3) were carefully compared with the values obtained via conventional weak-beam transmission electron microscopy studies. In addition, cluster analysis using atom probe tomography (APT) further demonstrated the potential of the WB-STEM for correlative electron tomography/APT experiments. © The Author 2017. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Entropy-Bayesian Inversion of Time-Lapse Tomographic GPR data for Monitoring Dielectric Permittivity and Soil Moisture Variations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hou, Z; Terry, N; Hubbard, S S

    2013-02-12

    In this study, we evaluate the possibility of monitoring soil moisture variation using tomographic ground penetrating radar travel time data through Bayesian inversion, which is integrated with entropy memory function and pilot point concepts, as well as efficient sampling approaches. It is critical to accurately estimate soil moisture content and variations in vadose zone studies. Many studies have illustrated the promise and value of GPR tomographic data for estimating soil moisture and associated changes, however, challenges still exist in the inversion of GPR tomographic data in a manner that quantifies input and predictive uncertainty, incorporates multiple data types, handles non-uniquenessmore » and nonlinearity, and honors time-lapse tomograms collected in a series. To address these challenges, we develop a minimum relative entropy (MRE)-Bayesian based inverse modeling framework that non-subjectively defines prior probabilities, incorporates information from multiple sources, and quantifies uncertainty. The framework enables us to estimate dielectric permittivity at pilot point locations distributed within the tomogram, as well as the spatial correlation range. In the inversion framework, MRE is first used to derive prior probability distribution functions (pdfs) of dielectric permittivity based on prior information obtained from a straight-ray GPR inversion. The probability distributions are then sampled using a Quasi-Monte Carlo (QMC) approach, and the sample sets provide inputs to a sequential Gaussian simulation (SGSim) algorithm that constructs a highly resolved permittivity/velocity field for evaluation with a curved-ray GPR forward model. The likelihood functions are computed as a function of misfits, and posterior pdfs are constructed using a Gaussian kernel. Inversion of subsequent time-lapse datasets combines the Bayesian estimates from the previous inversion (as a memory function) with new data. The memory function and pilot point design takes

  2. Entropy-Bayesian Inversion of Time-Lapse Tomographic GPR data for Monitoring Dielectric Permittivity and Soil Moisture Variations

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hou, Zhangshuan; Terry, Neil C.; Hubbard, Susan S.

    2013-02-22

    In this study, we evaluate the possibility of monitoring soil moisture variation using tomographic ground penetrating radar travel time data through Bayesian inversion, which is integrated with entropy memory function and pilot point concepts, as well as efficient sampling approaches. It is critical to accurately estimate soil moisture content and variations in vadose zone studies. Many studies have illustrated the promise and value of GPR tomographic data for estimating soil moisture and associated changes, however, challenges still exist in the inversion of GPR tomographic data in a manner that quantifies input and predictive uncertainty, incorporates multiple data types, handles non-uniquenessmore » and nonlinearity, and honors time-lapse tomograms collected in a series. To address these challenges, we develop a minimum relative entropy (MRE)-Bayesian based inverse modeling framework that non-subjectively defines prior probabilities, incorporates information from multiple sources, and quantifies uncertainty. The framework enables us to estimate dielectric permittivity at pilot point locations distributed within the tomogram, as well as the spatial correlation range. In the inversion framework, MRE is first used to derive prior probability density functions (pdfs) of dielectric permittivity based on prior information obtained from a straight-ray GPR inversion. The probability distributions are then sampled using a Quasi-Monte Carlo (QMC) approach, and the sample sets provide inputs to a sequential Gaussian simulation (SGSIM) algorithm that constructs a highly resolved permittivity/velocity field for evaluation with a curved-ray GPR forward model. The likelihood functions are computed as a function of misfits, and posterior pdfs are constructed using a Gaussian kernel. Inversion of subsequent time-lapse datasets combines the Bayesian estimates from the previous inversion (as a memory function) with new data. The memory function and pilot point design takes

  3. Overland flow dynamics through visual observation using time-lapse photographs

    NASA Astrophysics Data System (ADS)

    Silasari, Rasmiaditya; Blöschl, Günter

    2016-04-01

    Overland flow process on agricultural land is important to be investigated as it affects the stream discharge and water quality assessment. During rainfall events the formation of overland flow may happen through different processes (i.e. Hortonian or saturation excess overland flow) based on the governing soil hydraulic parameters (i.e. soil infiltration rate, soil water capacity). The dynamics of the soil water state and the processes will affect the surface runoff response which can be analyzed visually by observing the saturation patterns with a camera. Although visual observation was proven useful in laboratory experiments, the technique is not yet assessed for natural rainfall events. The aim of this work is to explore the use of time-lapse photographs of naturally occurring-saturation patterns in understanding the threshold processes of overland flow generation. The image processing produces orthographic projection of the saturation patterns which will be used to assess the dynamics of overland flow formation in relation with soil moisture state and rainfall magnitude. The camera observation was performed at Hydrological Open Air Laboratory (HOAL) catchment at Petzenkirchen, Lower Austria. The catchment covers an area of 66 ha dominated with agricultural land (87%). The mean annual precipitation and mean annual flow at catchment outlet are 750 mm and 4 l/s, respectively. The camera was set to observe the overland flow along a thalweg on an arable field which was drained in 1950s and has advantages of: (1) representing agricultural land as the dominant part of the catchment, (2) adjacent to the stream with clear visibility (no obstructing objects, such as trees), (3) drained area provides extra cases in understanding the response of tile drain outflow to overland flow formation and vice versa, and (4) in the vicinity of TDT soil moisture stations. The camera takes a picture with 1280 x 720 pixels resolution every minute and sends it directly in a PC via fiber

  4. Visualizing and quantifying movement from pre-recorded videos: The spectral time-lapse (STL) algorithm

    PubMed Central

    Madan, Christopher R

    2014-01-01

    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. PMID:25580219

  5. Quantitative force measurements in liquid using frequency modulation atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Uchihashi, Takayuki; Higgins, Michael J.; Yasuda, Satoshi; Jarvis, Suzanne P.; Akita, Seiji; Nakayama, Yoshikazu; Sader, John E.

    2004-10-01

    The measurement of short-range forces with the atomic force microscope (AFM) typically requires implementation of dynamic techniques to maintain sensitivity and stability. While frequency modulation atomic force microscopy (FM-AFM) is used widely for high-resolution imaging and quantitative force measurements in vacuum, quantitative force measurements using FM-AFM in liquids have proven elusive. Here we demonstrate that the formalism derived for operation in vacuum can also be used in liquids, provided certain modifications are implemented. To facilitate comparison with previous measurements taken using surface forces apparatus, we choose a model system (octamethylcyclotetrasiloxane) that is known to exhibit short-ranged structural ordering when confined between two surfaces. Force measurements obtained are found to be in excellent agreement with previously reported results. This study therefore establishes FM-AFM as a powerful tool for the quantitative measurement of forces in liquid.

  6. Time-lapse 3D electrical resistivity tomography to monitor soil-plant interactions

    NASA Astrophysics Data System (ADS)

    Boaga, Jacopo; Rossi, Matteo; Cassiani, Giorgio; Putti, Mario

    2013-04-01

    In this work we present the application of time-lapse non-invasive 3D micro- electrical tomography (ERT) to monitor soil-plant interactions in the root zone in the framework of the FP7 Project CLIMB (Climate Induced Changes on the Hydrology of Mediterranean Basins). The goal of the study is to gain a better understanding of the soil-vegetation interactions by the use of non-invasive techniques. We designed, built and installed a 3D electrical tomography apparatus for the monitoring of the root zone of a single apple tree in an orchard located in the Trentino region, Northern Italy. The micro-ERT apparatus consists of 48 buried electrodes on 4 instrumented micro boreholes plus 24 mini-electrodes on the surface spaced 0.1 m on a square grid. We collected repeated ERT and TDR soil moisture measurements for one year and performed two different controlled irrigation tests: one during a very dry Summer and one during a very wet and highly dynamic plant growing Spring period. We also ran laboratory analyses on soil specimens, in order to evaluate the electrical response at different saturation steps. The results demonstrate that 3D micro-ERT is capable of characterizing subsoil conditions and monitoring root zone activities, especially in terms of root zone suction regions. In particular, we note that in very dry conditions, 3D micro ERT can image water plumes in the shallow subsoil produced by a drip irrigation system. In the very dynamic growing season, under abundant irrigation, micro 3D ERT can detect the main suction zones caused by the tree root activity. Even though the quantitative use of this technique for moisture content balance suffers from well-known inversion difficulties, even the pure imaging of the active root zone is a valuable contribution. However the integration of the measurements in a fully coupled hydrogeophysical inversion is the way forward for a better understanding of subsoil interactions between biomass, hydrosphere and atmosphere.

  7. Distress Tolerance Treatment for Early-Lapse Smokers

    PubMed Central

    Brown, Richard A.; Palm, Kathleen M.; Strong, David R.; Lejuez, Carl W.; Kahler, Christopher W.; Zvolensky, Michael J.; Hayes, Steven C.; Wilson, Kelly G.; Gifford, Elizabeth V.

    2008-01-01

    A significant percentage of individuals attempting smoking cessation lapse within a matter of days, and very few are able to recover to achieve long-term abstinence. This observation suggests that many smokers may have quit-attempt histories characterized exclusively by early lapses to smoking following quit attempts. Recent negative-reinforcement conceptualizations of early lapse to smoking suggest that individuals' reactions to withdrawal and inability to tolerate the experience of these symptoms, rather than withdrawal severity itself, may represent an important treatment target in the development of new behavioral interventions for this subpopulation of smokers. This article presents the theoretical rationale and describes a novel, multicomponent distress-tolerance treatment for early-lapse smokers that incorporates behavioral and pharmacological elements of standard smoking-cessation treatment, whereas drawing distress-tolerance elements from exposure-based and Acceptance and Commitment Therapy–based treatment approaches. Preliminary data from a pilot study (N = 16) are presented, and clinical implications are discussed. PMID:18391050

  8. Developing an EEG-based on-line closed-loop lapse detection and mitigation system

    PubMed Central

    Wang, Yu-Te; Huang, Kuan-Chih; Wei, Chun-Shu; Huang, Teng-Yi; Ko, Li-Wei; Lin, Chin-Teng; Cheng, Chung-Kuan; Jung, Tzyy-Ping

    2014-01-01

    In America, 60% of adults reported that they have driven a motor vehicle while feeling drowsy, and at least 15–20% of fatal car accidents are fatigue-related. This study translates previous laboratory-oriented neurophysiological research to design, develop, and test an On-line Closed-loop Lapse Detection and Mitigation (OCLDM) System featuring a mobile wireless dry-sensor EEG headgear and a cell-phone based real-time EEG processing platform. Eleven subjects participated in an event-related lane-keeping task, in which they were instructed to manipulate a randomly deviated, fixed-speed cruising car on a 4-lane highway. This was simulated in a 1st person view with an 8-screen and 8-projector immersive virtual-reality environment. When the subjects experienced lapses or failed to respond to events during the experiment, auditory warning was delivered to rectify the performance decrements. However, the arousing auditory signals were not always effective. The EEG spectra exhibited statistically significant differences between effective and ineffective arousing signals, suggesting that EEG spectra could be used as a countermeasure of the efficacy of arousing signals. In this on-line pilot study, the proposed OCLDM System was able to continuously detect EEG signatures of fatigue, deliver arousing warning to subjects suffering momentary cognitive lapses, and assess the efficacy of the warning in near real-time to rectify cognitive lapses. The on-line testing results of the OCLDM System validated the efficacy of the arousing signals in improving subjects' response times to the subsequent lane-departure events. This study may lead to a practical on-line lapse detection and mitigation system in real-world environments. PMID:25352773

  9. Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy.

    PubMed

    You, Sixian; Tu, Haohua; Chaney, Eric J; Sun, Yi; Zhao, Youbo; Bower, Andrew J; Liu, Yuan-Zhi; Marjanovic, Marina; Sinha, Saurabh; Pu, Yang; Boppart, Stephen A

    2018-05-29

    Intravital microscopy (IVM) emerged and matured as a powerful tool for elucidating pathways in biological processes. Although label-free multiphoton IVM is attractive for its non-perturbative nature, its wide application has been hindered, mostly due to the limited contrast of each imaging modality and the challenge to integrate them. Here we introduce simultaneous label-free autofluorescence-multiharmonic (SLAM) microscopy, a single-excitation source nonlinear imaging platform that uses a custom-designed excitation window at 1110 nm and shaped ultrafast pulses at 10 MHz to enable fast (2-orders-of-magnitude improvement), simultaneous, and efficient acquisition of autofluorescence (FAD and NADH) and second/third harmonic generation from a wide array of cellular and extracellular components (e.g., tumor cells, immune cells, vesicles, and vessels) in living tissue using only 14 mW for extended time-lapse investigations. Our work demonstrates the versatility and efficiency of SLAM microscopy for tracking cellular events in vivo, and is a major enabling advance in label-free IVM.

  10. Visualization of conduit-matrix conductivity differences in a karst aquifer using time-lapse electrical resistivity

    NASA Astrophysics Data System (ADS)

    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.

  11. Selection of competent blastocysts for transfer by combining time-lapse monitoring and array CGH testing for patients undergoing preimplantation genetic screening: a prospective study with sibling oocytes

    PubMed Central

    2014-01-01

    Background Recent advances in time-lapse monitoring in IVF treatment have provided new morphokinetic markers for embryonic competence. However, there is still very limited information about the relationship between morphokinetic parameters, chromosomal compositions and implantation potential. Accordingly, this study aimed at investigating the effects of selecting competent blastocysts for transfer by combining time-lapse monitoring and array CGH testing on pregnancy and implantation outcomes for patients undergoing preimplantation genetic screening (PGS). Methods A total of 1163 metaphase II (MII) oocytes were retrieved from 138 PGS patients at a mean age of 36.6 ± 2.4 years. These sibling MII oocytes were then randomized into two groups after ICSI: 1) Group A, oocytes (n = 582) were cultured in the time-lapse system and 2) Group B, oocytes (n = 581) were cultured in the conventional incubator. For both groups, whole genomic amplification and array CGH testing were performed after trophectoderm biopsy on day 5. One to two euploid blastocysts within the most predictive morphokinetic parameters (Group A) or with the best morphological grade available (Group B) were selected for transfer to individual patients on day 6. Ongoing pregnancy and implantation rates were compared between the two groups. Results There were significant differences in clinical pregnancy rates between Group A and Group B (71.1% vs. 45.9%, respectively, p = 0.037). The observed implantation rate per embryo transfer significantly increased in Group A compared to Group B (66.2% vs. 42.4%, respectively, p = 0.011). Moreover, a significant increase in ongoing pregnancy rates was also observed in Group A compared to Group B (68.9% vs. 40.5%. respectively, p = 0.019). However, there was no significant difference in miscarriage rate between the time-lapse system and the conventional incubator (3.1% vs. 11.8%, respectively, p = 0.273). Conclusions This is the first prospective investigation using

  12. Highlights from the SoilCAM project: Soil Contamination, Advanced integrated characterisation and time-lapse Monitoring

    NASA Astrophysics Data System (ADS)

    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

  13. Time-lapse gravity and levelling in the sinkhole-endangered urban area of Bad Frankenhausen, Germany

    NASA Astrophysics Data System (ADS)

    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

  14. Cell segmentation in time-lapse fluorescence microscopy with temporally varying sub-cellular fusion protein patterns.

    PubMed

    Bunyak, Filiz; Palaniappan, Kannappan; Chagin, Vadim; Cardoso, M

    2009-01-01

    Fluorescently tagged proteins such as GFP-PCNA produce rich dynamically varying textural patterns of foci distributed in the nucleus. This enables the behavioral study of sub-cellular structures during different phases of the cell cycle. The varying punctuate patterns of fluorescence, drastic changes in SNR, shape and position during mitosis and abundance of touching cells, however, require more sophisticated algorithms for reliable automatic cell segmentation and lineage analysis. Since the cell nuclei are non-uniform in appearance, a distribution-based modeling of foreground classes is essential. The recently proposed graph partitioning active contours (GPAC) algorithm supports region descriptors and flexible distance metrics. We extend GPAC for fluorescence-based cell segmentation using regional density functions and dramatically improve its efficiency for segmentation from O(N(4)) to O(N(2)), for an image with N(2) pixels, making it practical and scalable for high throughput microscopy imaging studies.

  15. Lapse in Institutional Animal Care and Use Committee Continuing Reviews.

    PubMed

    Tsan, Min-Fu; Grabenbauer, Michael; Nguyen, Yen

    2016-01-01

    The United States federal animal welfare regulations and the Public Health Service Policy on Humane Care and Use of Laboratory Animals require that institutional animal care and use committees (IACUCs) conduct continuing reviews of all animal research activities. However, little is known about the lapse rate of IACUC continuing reviews, and how frequently investigators continue research activities during the lapse. It is also not clear what factors may contribute to an institution's lapse in IACUC continuing reviews. As part of the quality assurance program, the Department of Veterans Affairs (VA) has collected performance metric data for animal care and use programs since 2011. We analyzed IACUC continuing review performance data at 74-75 VA research facilities from 2011 through 2015. The IACUC continuing review lapse rates improved from 5.6% in 2011 to 2.7% in 2015. The rate of investigators continuing research activities during the lapse also decreased from 47.2% in 2012 to 7.4% in 2015. The type of IACUCs used and the size of animal research programs appeared to have no effect in facility's rates of lapse in IACUC continuing reviews. While approximately 80% of facilities reported no lapse in IACUC continuing reviews, approximately 14% of facilities had lapse rates of >10% each year. Some facilities appeared to be repeat offenders. Four facilities had IACUC lapse rates of >10% in at least 3 out of 5 years, suggesting a system problem in these facilities requiring remedial actions to improve their IACUC continuing review processes.

  16. Quantitative phase-digital holographic microscopy: a new imaging modality to identify original cellular biomarkers of diseases

    NASA Astrophysics Data System (ADS)

    Marquet, P.; Rothenfusser, K.; Rappaz, B.; Depeursinge, C.; Jourdain, P.; Magistretti, P. J.

    2016-03-01

    Quantitative phase microscopy (QPM) has recently emerged as a powerful label-free technique in the field of living cell imaging allowing to non-invasively measure with a nanometric axial sensitivity cell structure and dynamics. Since the phase retardation of a light wave when transmitted through the observed cells, namely the quantitative phase signal (QPS), is sensitive to both cellular thickness and intracellular refractive index related to the cellular content, its accurate analysis allows to derive various cell parameters and monitor specific cell processes, which are very likely to identify new cell biomarkers. Specifically, quantitative phase-digital holographic microscopy (QP-DHM), thanks to its numerical flexibility facilitating parallelization and automation processes, represents an appealing imaging modality to both identify original cellular biomarkers of diseases as well to explore the underlying pathophysiological processes.

  17. Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies.

    PubMed

    Sun, Wanxin; Chang, Shi; Tai, Dean C S; Tan, Nancy; Xiao, Guangfa; Tang, Huihuan; Yu, Hanry

    2008-01-01

    Liver fibrosis is associated with an abnormal increase in an extracellular matrix in chronic liver diseases. Quantitative characterization of fibrillar collagen in intact tissue is essential for both fibrosis studies and clinical applications. Commonly used methods, histological staining followed by either semiquantitative or computerized image analysis, have limited sensitivity, accuracy, and operator-dependent variations. The fibrillar collagen in sinusoids of normal livers could be observed through second-harmonic generation (SHG) microscopy. The two-photon excited fluorescence (TPEF) images, recorded simultaneously with SHG, clearly revealed the hepatocyte morphology. We have systematically optimized the parameters for the quantitative SHG/TPEF imaging of liver tissue and developed fully automated image analysis algorithms to extract the information of collagen changes and cell necrosis. Subtle changes in the distribution and amount of collagen and cell morphology are quantitatively characterized in SHG/TPEF images. By comparing to traditional staining, such as Masson's trichrome and Sirius red, SHG/TPEF is a sensitive quantitative tool for automated collagen characterization in liver tissue. Our system allows for enhanced detection and quantification of sinusoidal collagen fibers in fibrosis research and clinical diagnostics.

  18. Accurate virus quantitation using a Scanning Transmission Electron Microscopy (STEM) detector in a scanning electron microscope.

    PubMed

    Blancett, Candace D; Fetterer, David P; Koistinen, Keith A; Morazzani, Elaine M; Monninger, Mitchell K; Piper, Ashley E; Kuehl, Kathleen A; Kearney, Brian J; Norris, Sarah L; Rossi, Cynthia A; Glass, Pamela J; Sun, Mei G

    2017-10-01

    A method for accurate quantitation of virus particles has long been sought, but a perfect method still eludes the scientific community. Electron Microscopy (EM) quantitation is a valuable technique because it provides direct morphology information and counts of all viral particles, whether or not they are infectious. In the past, EM negative stain quantitation methods have been cited as inaccurate, non-reproducible, and with detection limits that were too high to be useful. To improve accuracy and reproducibility, we have developed a method termed Scanning Transmission Electron Microscopy - Virus Quantitation (STEM-VQ), which simplifies sample preparation and uses a high throughput STEM detector in a Scanning Electron Microscope (SEM) coupled with commercially available software. In this paper, we demonstrate STEM-VQ with an alphavirus stock preparation to present the method's accuracy and reproducibility, including a comparison of STEM-VQ to viral plaque assay and the ViroCyt Virus Counter. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  19. High speed quantitative digital microscopy

    NASA Technical Reports Server (NTRS)

    Castleman, K. R.; Price, K. H.; Eskenazi, R.; Ovadya, M. M.; Navon, M. A.

    1984-01-01

    Modern digital image processing hardware makes possible quantitative analysis of microscope images at high speed. This paper describes an application to automatic screening for cervical cancer. The system uses twelve MC6809 microprocessors arranged in a pipeline multiprocessor configuration. Each processor executes one part of the algorithm on each cell image as it passes through the pipeline. Each processor communicates with its upstream and downstream neighbors via shared two-port memory. Thus no time is devoted to input-output operations as such. This configuration is expected to be at least ten times faster than previous systems.

  20. Monitoring CO 2 sequestration into deep saline aquifer and associated salt intrusion using coupled multiphase flow modeling and time lapse electrical resistivity tomography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chuan Lu; CHI Zhang; Hai Hanag

    2014-04-01

    Successful geological storage and sequestration of carbon dioxide (CO2) require efficient monitoring of the migration of CO2 plume during and after large-scale injection in order to verify the containment of the injected CO2 within the target formation and to evaluate potential leakage risk. Field studies have shown that surface and cross-borehole electrical resistivity tomography (ERT) can be a useful tool in imaging and characterizing solute transport in heterogeneous subsurface. In this synthetic study, we have coupled a 3-D multiphase flow model with a parallel 3-D time-lapse ERT inversion code to explore the feasibility of using time-lapse ERT for simultaneously monitoringmore » the migration of CO2 plume in deep saline formation and potential brine intrusion into shallow fresh water aquifer. Direct comparisons of the inverted CO2 plumes resulting from ERT with multiphase flow simulation results indicate the ERT could be used to delineate the migration of CO2 plume. Detailed comparisons on the locations, sizes and shapes of CO2 plume and intruded brine plumes suggest that ERT inversion tends to underestimate the area review of the CO2 plume, but overestimate the thickness and total volume of the CO2 plume. The total volume of intruded brine plumes is overestimated as well. However, all discrepancies remain within reasonable ranges. Our study suggests that time-lapse ERT is a useful monitoring tool in characterizing the movement of injected CO2 into deep saline aquifer and detecting potential brine intrusion under large-scale field injection conditions.« less

  1. Improving time-lapse seismic repeatability: CO2CRC Otway site permanent geophone array field trials

    NASA Astrophysics Data System (ADS)

    Pevzner, Roman; Dupuis, Christian; Shulakova, Valeriya; Urosevic, Milovan; Lumley, David

    2013-04-01

    The proposed Stage 2C of the CO2CRC Otway project involves injection of a small amount (around 15,000 tonnes) of CO2/CH4 gas mixture into saline acquifer (Paaratte formation) at the depth of ~1.5 km. The seismic time-lapse signal will depend largely on the formation properties and the injection scenario, but is likely to be relatively weak. In order to improve time-lapse seismic monitoring capabilities by decreasing the noise level, a buried receiver arrays can be used. A small-scale trial of such an array was conducted at Otway site in June 2012. A set of 25 geophones was installed in 3 m deep boreholes in parallel to the same number of surface geophones. In addition, four geophones were placed into boreholes of 1 to 12 m depth. In order to assess the gain in the signal-to-noise ratio and repeatability, both active and passive seismic surveys were carried out. The surveys were conducted in relatively poor weather conditions, with rain, strong wind and thunderstorms increasing the noise level. We found that noise level for buried geophones is on average 20 dB lower compared to the surface ones. Furthermore, the combination of active and passive experiments has allowed us to perform a detailed classification of various noise sources. Acknowledgement The authors acknowledge the funding provided by the Australian government through its CRC program to support this CO2CRC research project. We also acknowledge the CO2CRC's corporate sponsors and the financial assistance provided through Australian National Low Emissions Coal Research and Development (ANLEC R&D). ANLEC R&D is supported by Australian Coal Association Low Emissions Technology Limited and the Australian Government through the Clean Energy Initiative.

  2. Computational method for multi-modal microscopy based on transport of intensity equation

    NASA Astrophysics Data System (ADS)

    Li, Jiaji; Chen, Qian; Sun, Jiasong; Zhang, Jialin; Zuo, Chao

    2017-02-01

    In this paper, we develop the requisite theory to describe a hybrid virtual-physical multi-modal imaging system which yields quantitative phase, Zernike phase contrast, differential interference contrast (DIC), and light field moment imaging simultaneously based on transport of intensity equation(TIE). We then give the experimental demonstration of these ideas by time-lapse imaging of live HeLa cell mitosis. Experimental results verify that a tunable lens based TIE system, combined with the appropriate post-processing algorithm, can achieve a variety of promising imaging modalities in parallel with the quantitative phase images for the dynamic study of cellular processes.

  3. Quantitative analysis of the effect of environmental-scanning electron microscopy on collagenous tissues.

    PubMed

    Lee, Woowon; Toussaint, Kimani C

    2018-05-31

    Environmental-scanning electron microscopy (ESEM) is routinely applied to various biological samples due to its ability to maintain a wet environment while imaging; moreover, the technique obviates the need for sample coating. However, there is limited research carried out on electron-beam (e-beam) induced tissue damage resulting from using the ESEM. In this paper, we use quantitative second-harmonic generation (SHG) microscopy to examine the effects of e-beam exposure from the ESEM on collagenous tissue samples prepared as either fixed, frozen, wet or dehydrated. Quantitative SHG analysis of tissues, before and after ESEM e-beam exposure in low-vacuum mode, reveals evidence of cross-linking of collagen fibers, however there are no structural differences observed in fixed tissue. Meanwhile wet-mode ESEM appears to radically alter the structure from a regular fibrous arrangement to a more random fiber orientation. We also confirm that ESEM images of collagenous tissues show higher spatial resolution compared to SHG microscopy, but the relative tradeoff with collagen specificity reduces its effectiveness in quantifying collagen fiber organization. Our work provides insight on both the limitations of the ESEM for tissue imaging, and the potential opportunity to use as a complementary technique when imaging fine features in the non-collagenous regions of tissue samples.

  4. Oufti: An integrated software package for high-accuracy, high-throughput quantitative microscopy analysis

    PubMed Central

    Paintdakhi, Ahmad; Parry, Bradley; Campos, Manuel; Irnov, Irnov; Elf, Johan; Surovtsev, Ivan; Jacobs-Wagner, Christine

    2016-01-01

    Summary With the realization that bacteria display phenotypic variability among cells and exhibit complex subcellular organization critical for cellular function and behavior, microscopy has re-emerged as a primary tool in bacterial research during the last decade. However, the bottleneck in today’s single-cell studies is quantitative image analysis of cells and fluorescent signals. Here, we address current limitations through the development of Oufti, a stand-alone, open-source software package for automated measurements of microbial cells and fluorescence signals from microscopy images. Oufti provides computational solutions for tracking touching cells in confluent samples, handles various cell morphologies, offers algorithms for quantitative analysis of both diffraction and non-diffraction-limited fluorescence signals, and is scalable for high-throughput analysis of massive datasets, all with subpixel precision. All functionalities are integrated in a single package. The graphical user interface, which includes interactive modules for segmentation, image analysis, and post-processing analysis, makes the software broadly accessible to users irrespective of their computational skills. PMID:26538279

  5. TRIIG - Time-lapse reproduction of images through interactive graphics. [digital processing of quality hard copy

    NASA Technical Reports Server (NTRS)

    Buckner, J. D.; Council, H. W.; Edwards, T. R.

    1974-01-01

    Description of the hardware and software implementing the system of time-lapse reproduction of images through interactive graphics (TRIIG). The system produces a quality hard copy of processed images in a fast and inexpensive manner. This capability allows for optimal development of processing software through the rapid viewing of many image frames in an interactive mode. Three critical optical devices are used to reproduce an image: an Optronics photo reader/writer, the Adage Graphics Terminal, and Polaroid Type 57 high speed film. Typical sources of digitized images are observation satellites, such as ERTS or Mariner, computer coupled electron microscopes for high-magnification studies, or computer coupled X-ray devices for medical research.

  6. Development of a Monitoring Method for Nonlabeled Human Pluripotent Stem Cell Growth by Time-Lapse Image Analysis.

    PubMed

    Suga, Mika; Kii, Hiroaki; Niikura, Keiichi; Kiyota, Yasujiro; Furue, Miho K

    2015-07-01

    : Cell growth is an important criterion for determining healthy cell conditions. When somatic cells or cancer cells are dissociated into single cells for passaging, the cell numbers can be counted at each passage, providing information on cell growth as an indicator of the health conditions of these cells. In the case of human pluripotent stem cells (hPSCs), because the cells are usually dissociated into cell clumps of ∼50-100 cells for passaging, cell counting is time-consuming. In the present study, using a time-lapse imaging system, we developed a method to determine the growth of hPSCs from nonlabeled live cell phase-contrast images without damaging these cells. Next, the hPSC colony areas and number of nuclei were determined and used to derive equations to calculate the cell number in hPSC colonies, which were assessed on time-lapse images acquired using a culture observation system. The relationships between the colony areas and nuclei numbers were linear, although the equation coefficients were dependent on the cell line used, colony size, colony morphology, and culture conditions. When the culture conditions became improper, the change in cell growth conditions could be detected by analysis of the phase-contrast images. This method provided real-time information on colony growth and cell growth rates without using treatments that can damage cells and could be useful for basic research on hPSCs and cell processing for hPSC-based therapy. This is the first study to use a noninvasive method using images to systemically determine the growth of human pluripotent stem cells (hPSCs) without damaging or wasting cells. This method would be useful for quality control during cell culture of clinical hPSCs. ©AlphaMed Press.

  7. [A time-lapse cinematographic analysis of ooplasmic movements during the cleavage ofPimpla turionellae L. (Hymenoptera)].

    PubMed

    Wolf, Rainer; Krause, Gerhard

    1971-09-01

    In the eggs ofPimpla turionellae, which are characterized by a long germ anlage ("long-germ egg" type), the cleavage nuclei primarily populate the anterior part and only later appear in the posterior of the egg lumen during the intravitelline cleavage. Gastrulation and segmentation also start within this anterior region. Time-lapse motion pictures served to observe and to check quantitatively even slow movements during cleavage and blastogenesis. In motion diagrams made by means of microkymographic technics the flow within the ooplasm along the longer axis of the egg has been timed.Shortly before the first cleavage in thestrictly unfertilized male eggs a short-time"unipolar flow" sets in from a primary initial region at 90% of their length. Thus a pillar of "central plasm" between both of the poles becomes shifted towards the posterior, while its outer coating layer of "marginal-plasm" is displaced forwards by the same distance. In eggs from fertilized females two successive flows of the same "unipolar" type have been observed.At the end of the third cleavage the energids, heretofore loosely grouped together, become distributed within the central plasm to form a "nuclear column". At the same time a fluently pulsatory "bipolar flow" sets in, within asecondary initial region at 80% of the egg length. Comparable to two mirror-image fountains, parts of the central plasm are carried towards the front pole and to the rear pole of the egg, respectively, while the marginal plasm, together with the oolemma, flows in opposite directions at times. With each pulsation the moving areas of the bipolar flow are shifted more and more towards the egg poles. The occurrence of bipolar flow pulsations, amounting to five, is correlated with the nuclear divisions in a still unknown way. In the rhythm of the bipolar flow, the energids become dispersed within the central plasm with a certain spatial lagging.After the bipolar flow has come to a halt, four further cleavages are indicated by

  8. Accurate Construction of Photoactivated Localization Microscopy (PALM) Images for Quantitative Measurements

    PubMed Central

    Coltharp, Carla; Kessler, Rene P.; Xiao, Jie

    2012-01-01

    Localization-based superresolution microscopy techniques such as Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) have allowed investigations of cellular structures with unprecedented optical resolutions. One major obstacle to interpreting superresolution images, however, is the overcounting of molecule numbers caused by fluorophore photoblinking. Using both experimental and simulated images, we determined the effects of photoblinking on the accurate reconstruction of superresolution images and on quantitative measurements of structural dimension and molecule density made from those images. We found that structural dimension and relative density measurements can be made reliably from images that contain photoblinking-related overcounting, but accurate absolute density measurements, and consequently faithful representations of molecule counts and positions in cellular structures, require the application of a clustering algorithm to group localizations that originate from the same molecule. We analyzed how applying a simple algorithm with different clustering thresholds (tThresh and dThresh) affects the accuracy of reconstructed images, and developed an easy method to select optimal thresholds. We also identified an empirical criterion to evaluate whether an imaging condition is appropriate for accurate superresolution image reconstruction with the clustering algorithm. Both the threshold selection method and imaging condition criterion are easy to implement within existing PALM clustering algorithms and experimental conditions. The main advantage of our method is that it generates a superresolution image and molecule position list that faithfully represents molecule counts and positions within a cellular structure, rather than only summarizing structural properties into ensemble parameters. This feature makes it particularly useful for cellular structures of heterogeneous densities and irregular geometries, and

  9. Fast, long-term, super-resolution imaging with Hessian structured illumination microscopy.

    PubMed

    Huang, Xiaoshuai; Fan, Junchao; Li, Liuju; Liu, Haosen; Wu, Runlong; Wu, Yi; Wei, Lisi; Mao, Heng; Lal, Amit; Xi, Peng; Tang, Liqiang; Zhang, Yunfeng; Liu, Yanmei; Tan, Shan; Chen, Liangyi

    2018-06-01

    To increase the temporal resolution and maximal imaging time of super-resolution (SR) microscopy, we have developed a deconvolution algorithm for structured illumination microscopy based on Hessian matrixes (Hessian-SIM). It uses the continuity of biological structures in multiple dimensions as a priori knowledge to guide image reconstruction and attains artifact-minimized SR images with less than 10% of the photon dose used by conventional SIM while substantially outperforming current algorithms at low signal intensities. Hessian-SIM enables rapid imaging of moving vesicles or loops in the endoplasmic reticulum without motion artifacts and with a spatiotemporal resolution of 88 nm and 188 Hz. Its high sensitivity allows the use of sub-millisecond excitation pulses followed by dark recovery times to reduce photobleaching of fluorescent proteins, enabling hour-long time-lapse SR imaging of actin filaments in live cells. Finally, we observed the structural dynamics of mitochondrial cristae and structures that, to our knowledge, have not been observed previously, such as enlarged fusion pores during vesicle exocytosis.

  10. Dual-modality wide-field photothermal quantitative phase microscopy and depletion of cell populations

    NASA Astrophysics Data System (ADS)

    Turko, Nir A.; Barnea, Itay; Blum, Omry; Korenstein, Rafi; Shaked, Natan T.

    2015-03-01

    We review our dual-modality technique for quantitative imaging and selective depletion of populations of cells based on wide-field photothermal (PT) quantitative phase imaging and simultaneous PT cell extermination. The cells are first labeled by plasmonic gold nanoparticles, which evoke local plasmonic resonance when illuminated by light in a wavelength corresponding to their specific plasmonic resonance peak. This reaction creates changes of temperature, resulting in changes of phase. This phase changes are recorded by a quantitative phase microscope (QPM), producing specific imaging contrast, and enabling bio-labeling in phase microscopy. Using this technique, we have shown discrimination of EGFR over-expressing (EGFR+) cancer cells from EGFR under-expressing (EGFR-) cancer cells. Then, we have increased the excitation power in order to evoke greater temperatures, which caused specific cell death, all under real-time phase acquisition using QPM. Close to 100% of all EGFR+ cells were immediately exterminated when illuminated with the strong excitation beam, while all EGFR- cells survived. For the second experiment, in order to simulate a condition where circulating tumor cells (CTCs) are present in blood, we have mixed the EGFR+ cancer cells with white blood cells (WBCs) from a healthy donor. Here too, we have used QPM to observe and record the phase of the cells as they were excited for selective visualization and then exterminated. The WBCs survival rate was over 95%, while the EGFR+ survival rate was under 5%. The technique may be the basis for real-time detection and controlled treatment of CTCs.

  11. Momentary changes in craving predict smoking lapse behavior: a laboratory study.

    PubMed

    Motschman, Courtney A; Germeroth, Lisa J; Tiffany, Stephen T

    2018-04-27

    Current research on factors that predict smoking lapse behavior is limited in its ability to fully characterize the critical moments leading up to decisions to smoke. We used a validated and widely used experimental analogue for smoking lapse to assess how moment-to-moment dynamics of craving relate to decisions to smoke. Heavy smokers (N = 128, M age = 35.9) participated in a 50-min laboratory delay to smoking task on 2 consecutive days, earning money for each 5 min they remained abstinent or ending the task by choosing to smoke. Participants rated craving and negative affect levels immediately prior to each choice. Participants were randomized to smoking as usual (n = 50) or overnight abstinence (n = 50 successfully abstained, n = 22 failed abstaining) prior to session 2. Discrete-time hazard models were used to examine craving and negative affect as time-varying predictors of smoking. Higher craving levels prior to smoking opportunities predicted increased risk of smoking. When controlling for craving levels, incremental increases in craving predicted increased smoking risk. Increases in negative affect incrementally predicted increased smoking risk at session 2 only. Smokers who failed to abstain were at a higher risk of smoking than those who successfully abstained, whereas abstinent and non-abstinent smokers did not differ in smoking risk. Findings demonstrate an extension of the smoking lapse paradigm that can be utilized to capture momentary changes in craving that predict smoking behavior. Evaluations of nuanced craving experiences may inform clinical and pharmacological research on preventing smoking lapse and relapse.

  12. Second harmonic generation quantitative measurements on collagen fibrils through correlation to electron microscopy

    NASA Astrophysics Data System (ADS)

    Bancelin, S.; Aimé, C.; Gusachenko, I.; Kowalczuk, L.; Latour, G.; Coradin, T.; Schanne-Klein, M.-C.

    2015-03-01

    Type I collagen is a major structural protein in mammals that shows highly structured macromolecular organizations specific to each tissue. This biopolymer is synthesized as triple helices, which self-assemble into fibrils (Ø =10-300 nm) and further form various 3D organization. In recent years, Second Harmonic Generation (SHG) microscopy has emerged as a powerful technique to probe in situ the fibrillar collagenous network within tissues. However, this optical technique cannot resolve most of the fibrils and is a coherent process, which has impeded quantitative measurements of the fibril diameter so far. In this study, we correlated SHG microscopy with Transmission Electron Microscopy to determine the sensitivity of SHG microscopy and to calibrate SHG signals as a function of the fibril diameter in reconstructed collagen gels. To that end, we synthetized isolated fibrils with various diameters and successfully imaged the very same fibrils with both techniques, down to 30 nm diameter. We observed that SHG signals scaled as the fourth power of the fibril diameter, as expected from analytical and numerical calculations. This calibration was then applied to diabetic rat cornea in which we successfully recovered the diameter of hyperglycemia-induced fibrils in the Descemet's membrane without having to resolve them. Finally we derived the first hyperpolarizability from a single collagen triple helix which validates the bottom-up approach used to calculate the non-linear response at the fibrillar scale and denotes a parallel alignment of triple helices within the fibrils. These results represent a major step towards quantitative SHG imaging of nm-sized collagen fibrils.

  13. Quantitative investigation of red blood cell three-dimensional geometric and chemical changes in the storage lesion using digital holographic microscopy.

    PubMed

    Jaferzadeh, Keyvan; Moon, Inkyu

    2015-11-01

    Quantitative phase information obtained by digital holographic microscopy (DHM) can provide new insight into the functions and morphology of single red blood cells (RBCs). Since the functionality of a RBC is related to its three-dimensional (3-D) shape, quantitative 3-D geometric changes induced by storage time can help hematologists realize its optimal functionality period. We quantitatively investigate RBC 3-D geometric changes in the storage lesion using DHM. Our experimental results show that the substantial geometric transformation of the biconcave-shaped RBCs to the spherocyte occurs due to RBC storage lesion. This transformation leads to progressive loss of cell surface area, surface-to-volume ratio, and functionality of RBCs. Furthermore, our quantitative analysis shows that there are significant correlations between chemical and morphological properties of RBCs.

  14. Optical Flow Applied to Time-Lapse Image Series to Estimate Glacier Motion in the Southern Patagonia Ice Field

    NASA Astrophysics Data System (ADS)

    Lannutti, E.; Lenzano, M. G.; Toth, C.; Lenzano, L.; Rivera, A.

    2016-06-01

    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.

  15. Quantitative intact specimen magnetic resonance microscopy at 3.0 T.

    PubMed

    Bath, Kevin G; Voss, Henning U; Jing, Deqiang; Anderson, Stewart; Hempstead, Barbara; Lee, Francis S; Dyke, Jonathan P; Ballon, Douglas J

    2009-06-01

    In this report, we discuss the application of a methodology for high-contrast, high-resolution magnetic resonance microscopy (MRM) of murine tissue using a 3.0-T imaging system. We employed a threefold strategy that included customized specimen preparation to maximize image contrast, three-dimensional data acquisition to minimize scan time and custom radiofrequency resonator design to maximize signal sensitivity. Images had a resolution of 100 x 78 x 78 microm(3) with a signal-to-noise ratio per voxel greater than 25:1 and excellent contrast-to-noise ratios over a 30-min acquisition. We quantitatively validated the methods through comparisons of neuroanatomy across two lines of genetically engineered mice. Specifically, we were able to detect volumetric differences of as little as 9% between genetically engineered mouse strains in multiple brain regions that were predictive of underlying impairments in brain development. The overall methodology was straightforward to implement and provides ready access to basic MRM at field strengths that are widely available in both the laboratory and the clinic.

  16. Quantitative evaluation of software packages for single-molecule localization microscopy.

    PubMed

    Sage, Daniel; Kirshner, Hagai; Pengo, Thomas; Stuurman, Nico; Min, Junhong; Manley, Suliana; Unser, Michael

    2015-08-01

    The quality of super-resolution images obtained by single-molecule localization microscopy (SMLM) depends largely on the software used to detect and accurately localize point sources. In this work, we focus on the computational aspects of super-resolution microscopy and present a comprehensive evaluation of localization software packages. Our philosophy is to evaluate each package as a whole, thus maintaining the integrity of the software. We prepared synthetic data that represent three-dimensional structures modeled after biological components, taking excitation parameters, noise sources, point-spread functions and pixelation into account. We then asked developers to run their software on our data; most responded favorably, allowing us to present a broad picture of the methods available. We evaluated their results using quantitative and user-interpretable criteria: detection rate, accuracy, quality of image reconstruction, resolution, software usability and computational resources. These metrics reflect the various tradeoffs of SMLM software packages and help users to choose the software that fits their needs.

  17. Automated analysis of time-lapse fluorescence microscopy images: from live cell images to intracellular foci.

    PubMed

    Dzyubachyk, Oleh; Essers, Jeroen; van Cappellen, Wiggert A; Baldeyron, Céline; Inagaki, Akiko; Niessen, Wiro J; Meijering, Erik

    2010-10-01

    Complete, accurate and reproducible analysis of intracellular foci from fluorescence microscopy image sequences of live cells requires full automation of all processing steps involved: cell segmentation and tracking followed by foci segmentation and pattern analysis. Integrated systems for this purpose are lacking. Extending our previous work in cell segmentation and tracking, we developed a new system for performing fully automated analysis of fluorescent foci in single cells. The system was validated by applying it to two common tasks: intracellular foci counting (in DNA damage repair experiments) and cell-phase identification based on foci pattern analysis (in DNA replication experiments). Experimental results show that the system performs comparably to expert human observers. Thus, it may replace tedious manual analyses for the considered tasks, and enables high-content screening. The described system was implemented in MATLAB (The MathWorks, Inc., USA) and compiled to run within the MATLAB environment. The routines together with four sample datasets are available at http://celmia.bigr.nl/. The software is planned for public release, free of charge for non-commercial use, after publication of this article.

  18. Real-time intravital imaging of pH variation associated with osteoclast activity.

    PubMed

    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.

  19. A low-cost landslide displacement activity assessment from time-lapse photogrammetry and rainfall data: Application to the Tessina landslide site

    NASA Astrophysics Data System (ADS)

    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

  20. Ptychography: use of quantitative phase information for high-contrast label free time-lapse imaging of living cells

    NASA Astrophysics Data System (ADS)

    Suman, Rakesh; O'Toole, Peter

    2014-03-01

    Here we report a novel label free, high contrast and quantitative method for imaging live cells. The technique reconstructs an image from overlapping diffraction patterns using a ptychographical algorithm. The algorithm utilises both amplitude and phase data from the sample to report on quantitative changes related to the refractive index (RI) and thickness of the specimen. We report the ability of this technique to generate high contrast images, to visualise neurite elongation in neuronal cells, and to provide measure of cell proliferation.

  1. Quantifying receptor trafficking and colocalization with confocal microscopy.

    PubMed

    Pike, Jeremy A; Styles, Iain B; Rappoport, Joshua Z; Heath, John K

    2017-02-15

    Confocal microscopy is a powerful tool for the study of cellular receptor trafficking and endocytosis. Unbiased and robust image analysis workflows are required for the identification, and study, of aberrant trafficking. After a brief review of related strategies, identifying both good and bad practice, custom workflows for the analysis of live cell 3D time-lapse data are presented. Strategies for data pre-processing, including denoising and background subtraction are considered. We use a 3D level set protocol to accurately segment cells using only the signal from fluorescently labelled receptor. A protocol for the quantification of changes to subcellular receptor distribution over time is then presented. As an example, ligand stimulated trafficking of epidermal growth factor receptor (EGFR) is shown to be significantly reduced in both AG1478 and Dynasore treated cells. Protocols for the quantitative analysis of colocalization between receptor and endosomes are also introduced, including strategies for signal isolation and statistical testing. By calculating the Manders and Pearson coefficients, both co-occurrence and correlation can be assessed. A statistically significant decrease in the level of ligand induced co-occurrence between EGFR and rab5 positive endosomes is demonstrated for both the AG1478 and Dynasore treated cells relative to a control. Finally, a strategy for the visualisation of co-occurrence is presented, which provides an unbiased alternative to colour overlays. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  2. Diagnosis of aerobic vaginitis by quantitative real-time PCR.

    PubMed

    Rumyantseva, T A; Bellen, G; Savochkina, Y A; Guschin, A E; Donders, G G G

    2016-07-01

    To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. Some level of AV was diagnosed in 23 (23.7 %) cases. Various concentrations of Enterobacteriacea, Staphylococcus spp., Streptococcus spp. were detected an all patients. Enterococcus spp. were detected in 76 (78.3 %) cases. Summarized concentrations of aerobes were tenfold higher in AV-positive compared to AV-negative cases [7.30lg vs 6.06lg (p = 0.02)]. Concentrations of aerobes in severe, moderate and light AV cases did not vary significantly (p = 0.14). Concentration of lactobacilli was 1000-fold lower in AV-positive cases compared to normal cases (5.3lg vs 8.3lg, p < 0.0001). Streptococcus spp. dominated in the majority of AV-positive cases [19/22 (86.4 %) samples]. The relation of high loads of aerobes to the low numbers of Lactobacilli are a reliable marker for the presence of AV and could substitute microscopy as a test. PCR may be a good standardized substitution for AV diagnosis in settings where well-trained microscopists are lacking.

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

    NASA Astrophysics Data System (ADS)

    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

  4. Cross-sectional and Longitudinal Analyses of Everyday Memory Lapses in Older Adults

    PubMed Central

    McAlister, Courtney; Schmitter-Edgecombe, Maureen

    2016-01-01

    Everyday memory lapses experienced by older adults (OA) were examined using a daily-diary checklist and retrospective questionnaire. In Experiment 1, 138 younger and 138 OAs indicated the frequency of forgetting of 16 memory lapses, and whether each occurred daily during the course of a week. OAs reported more memory lapses on the questionnaire, but not the daily diary. OAs reported more frequently forgetting names and words, while younger adults had more difficulty with appointments and personal dates. Fewer memory lapses on the daily diary were related to better performance on a laboratory-memory measure for OAs. In Experiment 2, 62 OAs returned for a five-year follow-up and endorsed experiencing more memory lapses on the daily diary compared to baseline, specifically forgetting more names and words, but not the retrospective questionnaire. Daily checklist memory lapses again correlated with the laboratory-memory measure. A daily checklist may be a viable way to assess everyday memory lapses. PMID:26810777

  5. Prevalence of human immunodeficiency virus RNA and antibody in first-time, lapsed, and repeat blood donations across five international regions and relative efficacy of alternative screening scenarios.

    PubMed

    Bruhn, Roberta; Lelie, Nico; Custer, Brian; Busch, Michael; Kleinman, Steven

    2013-10-01

    Twenty-one blood organizations from five geographical regions provided HIV individual donation (ID)-NAT and serology data on 11,787,610 donations. Infections were classified as anti-HIV-/RNA+ window period (WP), anti-HIV+/RNA+ concordant positive (CP) or anti-HIV+/RNA- elite controller (EC). Residual risk and efficacy of several screening scenarios were estimated for first time, lapsed and repeat donations. WP residual risk estimates assumed a 50% infectious dose of 3.16 virions and a 50% detection limit of 2.7 HIV RNA copies/mL for ID-NAT and 10,000 copies/mL for p24Ag. Infectivity for CP (100%) and EC (2.2%) donations was estimated based on viral load distributions and 100-fold reduced infectivity by antibody neutralization as reported elsewhere. Efficacy was calculated as proportion of transmission risk removed from baseline (i.e. in absence of any screening). There was no significant difference in transmission risk between lapsed and repeat donations in any region. Risk was 3.8-fold higher in first time than combined lapsed/repeat donations in South Africa but not in other regions. Screening strategies were most efficacious at interdicting infectious transfusions in first time (98.7-99.8%) followed by lapsed (97.6-99.7%) and repeat (86.8-97.7%) donations in all regions combined. In each donor category the efficacy of ID-NAT alone (97.7-99.8%) was superior to that of minipool (MP)-NAT/anti-HIV (95.0-99.6%) and p24 Ag/anti-HIV (89.8-99.1%). Efficacy patterns were similar by donor/donation status in each region despite large differences in HIV prevalence and transmission risk. As similar data become available for HBV and HCV, this modeling may be useful in cost effectiveness analyses of alternative testing scenarios. © 2013 American Association of Blood Banks.

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

    USGS Publications Warehouse

    Karacan, C. Özgen; Olea, Ricardo A.

    2013-01-01

    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. Furthermore, performing filter simulations using point-wise data and TIs could be used to predict methane quantity in coal seams subjected to degasification. During the course of the study, it was shown that the material balance of gas produced by wellbores and the GIP reductions in coal seams predicted using filter simulations compared very well, showing the success of filter simulations for continuous variables in this case study. Quantitative results from filter simulations of GIP within the studied area briefly showed that GIP was reduced from an initial ∼73 Bcf (median) to ∼46 Bcf (2011), representing a 37 % decrease and varying spatially through degasification. It is forecasted that there will be an additional ∼2 Bcf reduction in methane quantity between 2011 and 2015. This study and presented results showed that the applied methodology and utilized techniques can be used to map GIP and its change within coal seams after degasification, which can further be used for ventilation design for methane control in coal mines.

  7. Near-surface temperature lapse rates in a mountainous catchment in the Chilean Andes

    NASA Astrophysics Data System (ADS)

    Ayala; Schauwecker, S.; Pellicciotti, F.; McPhee, J. P.

    2011-12-01

    In mountainous areas, and in the Chilean Andes in particular, the irregular and sparse distribution of recording stations resolves insufficiently the variability of climatic factors such as precipitation, temperature and relative humidity. Assumptions about air temperature variability in space and time have a strong effect on the performance of hydrologic models that represent snow processes such as accumulation and ablation. These processes have large diurnal variations, and assumptions that average over longer time periods (days, weeks or months) may reduce the predictive capacity of these models under different climatic conditions from those for which they were calibrated. They also introduce large uncertainties when such models are used to predict processes with strong subdiurnal variability such as snowmelt dynamics. In many applications and modeling exercises, temperature is assumed to decrease linearly with elevation, using the free-air moist adiabatic lapse rate (MALR: 0.0065°C/m). Little evidence is provided for this assumption, however, and recent studies have shown that use of lapse rates that are uniform in space and constant in time is not appropriate. To explore the validity of this approach, near-surface (2 m) lapse rates were calculated and analyzed at different temporal resolution, based on a new data set of spatially distributed temperature sensors setup in a high elevation catchment of the dry Andes of Central Chile (approx. 33°S). Five minutes temperature data were collected between January 2011 and April 2011 in the Ojos de Agua catchment, using two Automatic Weather Stations (AWSs) and 13 T-loggers (Hobo H8 Pro Temp with external data logger), ranging in altitude from 2230 to 3590 m.s.l.. The entire catchment was snow free during our experiment. We use this unique data set to understand the main controls over temperature variability in time and space, and test whether lapse rates can be used to describe the spatial variations of air

  8. Silver nanoparticle-induced degranulation observed with quantitative phase microscopy

    NASA Astrophysics Data System (ADS)

    Yang, Wenzhong; Lee, Seungrag; Lee, Jiyong; Bae, Yoonsung; Kim, Dugyoung

    2010-07-01

    Monitoring a degranulation process in a live mast cell is a quite important issue in immunology and pharmacology. Because the size of a granule is normally much smaller than the resolution limit of an optical microscope system, there is no direct real-time live cell imaging technique for observing degranulation processes except for fluorescence imaging techniques. In this research, we propose optical quantitative phase microscopy (QPM) as a new observation tool to study degranulation processes in a live mast cell without any fluorescence labeling. We measure the cell volumes and the cross sectional profiles (x-z plane) of an RBL-2H3 cell and a HeLa cell, before and after they are exposed to calcium ionophore A23187 and silver nanoparticles (AgNPs). We verify that the volume and the cross sectional line profile of the RBL-2H3 cell were changed significantly when it was exposed to A23187. When 50 μg/mL of AgNP is used instead of A23187, the measurements of cell volume and cross sectional profiles indicate that RBL-2H3 cells also follow degranulation processes. Degranulation processes for these cells are verified by monitoring the increase of intracellular calcium ([Ca2+]i) and histamine with fluorescent methods.

  9. Development of Nomarski microscopy for quantitative determination of surface topography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hartman, J. S.; Gordon, R. L.; Lessor, D. L.

    1979-01-01

    The use of Nomarski differential interference contrast (DIC) microscopy has been extended to provide nondestructive, quantitative analysis of a sample's surface topography. Theoretical modeling has determined the dependence of the image intensity on the microscope's optical components, the sample's optical properties, and the sample's surface orientation relative to the microscope. Results include expressions to allow the inversion of image intensity data to determine sample surface slopes. A commercial Nomarski system has been modified and characterized to allow the evaluation of the optical model. Data have been recorded with smooth, planar samples that verify the theoretical predictions.

  10. Multiplex real-time quantitative PCR, microscopy and rapid diagnostic immuno-chromatographic tests for the detection of Plasmodium spp: performance, limit of detection analysis and quality assurance.

    PubMed

    Khairnar, Krishna; Martin, Donald; Lau, Rachel; Ralevski, Filip; Pillai, Dylan R

    2009-12-09

    Accurate laboratory diagnosis of malaria species in returning travelers is paramount in the treatment of this potentially fatal infectious disease. A total of 466 blood specimens from returning travelers to Africa, Asia, and South/Central America with suspected malaria infection were collected between 2007 and 2009 at the reference public health laboratory. These specimens were assessed by reference microscopy, multipex real-time quantitative polymerase chain reaction (QPCR), and two rapid diagnostic immuno-chromatographic tests (ICT) in a blinded manner. Key clinical laboratory parameters such as limit of detection (LOD) analysis on clinical specimens by parasite stage, inter-reader variability of ICTs, staffing implications, quality assurance and cost analysis were evaluated. QPCR is the most analytically sensitive method (sensitivity 99.41%), followed by CARESTART (sensitivity 88.24%), and BINAXNOW (sensitivity 86.47%) for the diagnosis of malaria in returning travelers when compared to reference microscopy. However, microscopy was unable to specifically identify Plasmodia spp. in 18 out of 170 positive samples by QPCR. Moreover, the 17 samples that were negative by microscopy and positive by QPCR were also positive by ICTs. Quality assurance was achieved for QPCR by exchanging a blinded proficiency panel with another reference laboratory. The Kappa value of inter-reader variability among three readers for BINAXNOW and CARESTART was calculated to be 0.872 and 0.898 respectively. Serial dilution studies demonstrated that the QPCR cycle threshold correlates linearly with parasitemia (R(2) = 0.9746) in a clinically relevant dynamic range and retains a LOD of 11 rDNA copies/microl for P. falciparum, which was several log lower than reference microscopy and ICTs. LOD for QPCR is affected not only by parasitemia but the parasite stage distribution of each clinical specimen. QPCR was approximately 6-fold more costly than reference microscopy. These data suggest that

  11. Automated Quantitative Rare Earth Elements Mineralogy by Scanning Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Sindern, Sven; Meyer, F. Michael

    2016-09-01

    Increasing industrial demand of rare earth elements (REEs) stems from the central role they play for advanced technologies and the accelerating move away from carbon-based fuels. However, REE production is often hampered by the chemical, mineralogical as well as textural complexity of the ores with a need for better understanding of their salient properties. This is not only essential for in-depth genetic interpretations but also for a robust assessment of ore quality and economic viability. The design of energy and cost-efficient processing of REE ores depends heavily on information about REE element deportment that can be made available employing automated quantitative process mineralogy. Quantitative mineralogy assigns numeric values to compositional and textural properties of mineral matter. Scanning electron microscopy (SEM) combined with a suitable software package for acquisition of backscatter electron and X-ray signals, phase assignment and image analysis is one of the most efficient tools for quantitative mineralogy. The four different SEM-based automated quantitative mineralogy systems, i.e. FEI QEMSCAN and MLA, Tescan TIMA and Zeiss Mineralogic Mining, which are commercially available, are briefly characterized. Using examples of quantitative REE mineralogy, this chapter illustrates capabilities and limitations of automated SEM-based systems. Chemical variability of REE minerals and analytical uncertainty can reduce performance of phase assignment. This is shown for the REE phases parisite and synchysite. In another example from a monazite REE deposit, the quantitative mineralogical parameters surface roughness and mineral association derived from image analysis are applied for automated discrimination of apatite formed in a breakdown reaction of monazite and apatite formed by metamorphism prior to monazite breakdown. SEM-based automated mineralogy fulfils all requirements for characterization of complex unconventional REE ores that will become

  12. Imaging cell biology in live animals: ready for prime time.

    PubMed

    Weigert, Roberto; Porat-Shliom, Natalie; Amornphimoltham, Panomwat

    2013-06-24

    Time-lapse fluorescence microscopy is one of the main tools used to image subcellular structures in living cells. Yet for decades it has been applied primarily to in vitro model systems. Thanks to the most recent advancements in intravital microscopy, this approach has finally been extended to live rodents. This represents a major breakthrough that will provide unprecedented new opportunities to study mammalian cell biology in vivo and has already provided new insight in the fields of neurobiology, immunology, and cancer biology.

  13. Mathematical analysis of endothelial sibling pair cell-cell interactions using time-lapse cinematography data.

    PubMed

    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. Quantitative assessment of neural outgrowth using spatial light interference microscopy

    NASA Astrophysics Data System (ADS)

    Lee, Young Jae; Cintora, Pati; Arikkath, Jyothi; Akinsola, Olaoluwa; Kandel, Mikhail; Popescu, Gabriel; Best-Popescu, Catherine

    2017-06-01

    Optimal growth as well as branching of axons and dendrites is critical for the nervous system function. Neuritic length, arborization, and growth rate determine the innervation properties of neurons and define each cell's computational capability. Thus, to investigate the nervous system function, we need to develop methods and instrumentation techniques capable of quantifying various aspects of neural network formation: neuron process extension, retraction, stability, and branching. During the last three decades, fluorescence microscopy has yielded enormous advances in our understanding of neurobiology. While fluorescent markers provide valuable specificity to imaging, photobleaching, and photoxicity often limit the duration of the investigation. Here, we used spatial light interference microscopy (SLIM) to measure quantitatively neurite outgrowth as a function of cell confluence. Because it is label-free and nondestructive, SLIM allows for long-term investigation over many hours. We found that neurons exhibit a higher growth rate of neurite length in low-confluence versus medium- and high-confluence conditions. We believe this methodology will aid investigators in performing unbiased, nondestructive analysis of morphometric neuronal parameters.

  15. Time-Lapse Micro-Tomography Measurements and Determination of Effective Transport Properties of Snow Metamorphism Under Advective Conditions

    NASA Astrophysics Data System (ADS)

    Ebner, P. P.; Grimm, S.; Steen-Larsen, H. C.; Schneebeli, M.; Steinfeld, A.

    2014-12-01

    The metamorphism of snow under advective air flow, with and without temperature gradient, was never experimentally investigated. We developed a new sample holder where metamorphism under advective conditions can be observed and measured using time-lapse micro-tomography [1]. Long-term experiments were performed and direct pore-level simulation (DPLS) [2,3] was directly applied on the extracted 3D digital geometry of the snow to calculate the effective transport properties by solving the governing fluid flow equations. The results showed no effect of isothermal advection, compared to rates typical for isothermal metamorphism. Appling a temperature gradient, the results showed increased snow metamorphism compared to rates typical for temperature gradient metamorphism. However, for both cases a change in the isotopic composition in the air as well as in the snow sample could be observed. These measurements could be influential to better understand snow-air exchange processes relevant for atmospheric chemistry and isotopic composition. REFERENCES[1] Ebner P. P., Grimm S., Schneebeli M., and Steinfeld A.: An instrumented sample holder for time-lapse micro-tomography measurements of snow under advective airflow. Geoscientific Instrumentation, Methods and Data Systems 4(2014), 353-373. [2] Zermatten E., Haussener S., Schneebeli M., and Steinfeld A.: Tomography-based determination of permeability and Dupuit-Forchheimer coefficient of characteristic snow samples. Journal of Glaciology 57(2011), 811-816. [3] Zermatten E., Schneebeli M., Arakawa H., and Steinfeld A.: Tomography-based determination of porosity, specific area and permeability of snow and comparison with measurements. Cold Regions Science and Technology 97 (2014), 33-40. Fig. 1: 3-D surface rendering of a refrozen wet snow sample with fluid flow streamline.

  16. Global dynamics of selective attention and its lapses in primary auditory cortex.

    PubMed

    Lakatos, Peter; Barczak, Annamaria; Neymotin, Samuel A; McGinnis, Tammy; Ross, Deborah; Javitt, Daniel C; O'Connell, Monica Noelle

    2016-12-01

    Previous research demonstrated that while selectively attending to relevant aspects of the external world, the brain extracts pertinent information by aligning its neuronal oscillations to key time points of stimuli or their sampling by sensory organs. This alignment mechanism is termed oscillatory entrainment. We investigated the global, long-timescale dynamics of this mechanism in the primary auditory cortex of nonhuman primates, and hypothesized that lapses of entrainment would correspond to lapses of attention. By examining electrophysiological and behavioral measures, we observed that besides the lack of entrainment by external stimuli, attentional lapses were also characterized by high-amplitude alpha oscillations, with alpha frequency structuring of neuronal ensemble and single-unit operations. Entrainment and alpha-oscillation-dominated periods were strongly anticorrelated and fluctuated rhythmically at an ultra-slow rate. Our results indicate that these two distinct brain states represent externally versus internally oriented computational resources engaged by large-scale task-positive and task-negative functional networks.

  17. Reconsidering the Device in the Drawer: Lapses as a Design Opportunity in Personal Informatics

    PubMed Central

    Epstein, Daniel A.; Kang, Jennifer H.; Pina, Laura R.; Fogarty, James; Munson, Sean A.

    2017-01-01

    People stop using personal tracking tools over time, referred to as the lapsing stage of their tool use. We explore how designs can support people when they lapse in tracking, considering how to design data representations for a person who lapses in Fitbit use. Through a survey of 141 people who had lapsed in using Fitbit, we identified three use patterns and four perspectives on tracking. Participants then viewed seven visual representations of their Fitbit data and seven approaches to framing this data. Participant Fitbit use and perspective on tracking influenced their preference, which we surface in a series of contrasts. Specifically, our findings guide selecting appropriate aggregations from Fitbit use (e.g., aggregate more when someone has less data), choosing an appropriate framing technique from tracking perspective (e.g., ensure framing aligns with how the person feels about tracking), and creating appropriate social comparisons (e.g., portray the person positively compared to peers). We conclude by discussing how these contrasts suggest new designs and opportunities in other tracking domains. PMID:28516173

  18. Time-lapse changes of P- and S-wave velocities and shear wave splitting in the first year after the 2011 Tohoku earthquake, Japan: shallow subsurface

    NASA Astrophysics Data System (ADS)

    Sawazaki, Kaoru; Snieder, Roel

    2013-04-01

    We detect time-lapse changes in P- and S-wave velocities (hereafter, VP and VS, respectively) and shear wave splitting parameters associated with the 2011 Tohoku earthquake, Japan, at depths between 0 and 504 m. We estimate not only medium parameters but also the 95 per cent confidence interval of the estimated velocity change by applying a new least squares inversion scheme to the deconvolution analysis of KiK-net vertical array records. Up to 6 per cent VS reduction is observed at more than half of the analysed KiK-net stations in northeastern Japan with over 95 per cent confidence in the first month after the main shock. There is a considerable correlation between the S-wave traveltime delay and the maximum horizontal dynamic strain (MDS) by the main shock motion when the strain exceeds 5 × 10- 4 on the ground surface. This correlation is not clearly observed for MDS at the borehole bottom. On the contrary, VP and shear wave splitting parameters do not show systematic changes after the Tohoku earthquake. These results indicate that the time-lapse change is concentrated near the ground surface, especially in loosely packed soil layers. We conclude that the behaviour of VP, VS and shear wave splitting parameters are explained by the generation of omnidirectional cracks near the ground surface and by the diffusion of water in the porous subsurface. Recovery of VS should be related to healing of the crack which is proportional to the logarithm of the lapse time after the main shock and/or to decompaction after shaking.

  19. Soundscape and Noise Exposure Monitoring in a Marine Protected Area Using Shipping Data and Time-Lapse Footage.

    PubMed

    Merchant, Nathan D; Pirotta, Enrico; Barton, Tim R; Thompson, Paul M

    2016-01-01

    We review recent work that developed new techniques for underwater noise assessment that integrate acoustic monitoring with automatic identification system (AIS) shipping data and time-lapse video, meteorological, and tidal data. Two sites were studied within the Moray Firth Special Area of Conservation (SAC) for bottlenose dolphins, where increased shipping traffic is expected from construction of offshore wind farms outside the SAC. Noise exposure varied markedly between the sites, and natural and anthropogenic contributions were characterized using multiple data sources. At one site, AIS-operating vessels accounted for total cumulative sound exposure (0.1-10 kHz), suggesting that noise modeling using the AIS would be feasible.

  20. High-resolution quantitative determination of dielectric function by using scattering scanning near-field optical microscopy

    PubMed Central

    Tranca, D. E.; Stanciu, S. G.; Hristu, R.; Stoichita, C.; Tofail, S. A. M.; Stanciu, G. A.

    2015-01-01

    A new method for high-resolution quantitative measurement of the dielectric function by using scattering scanning near-field optical microscopy (s-SNOM) is presented. The method is based on a calibration procedure that uses the s-SNOM oscillating dipole model of the probe-sample interaction and quantitative s-SNOM measurements. The nanoscale capabilities of the method have the potential to enable novel applications in various fields such as nano-electronics, nano-photonics, biology or medicine. PMID:26138665

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

    PubMed

    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.

  2. Can arousing feedback rectify lapses in driving? Prediction from EEG power spectra.

    PubMed

    Lin, Chin-Teng; Huang, Kuan-Chih; Chuang, Chun-Hsiang; Ko, Li-Wei; Jung, Tzyy-Ping

    2013-10-01

    This study explores the neurophysiological changes, measured using an electroencephalogram (EEG), in response to an arousing warning signal delivered to drowsy drivers, and predicts the efficacy of the feedback based on changes in the EEG. Eleven healthy subjects participated in sustained-attention driving experiments. The driving task required participants to maintain their cruising position and compensate for randomly induced lane deviations using the steering wheel, while their EEG and driving performance were continuously monitored. The arousing warning signal was delivered to participants who experienced momentary behavioral lapses, failing to respond rapidly to lane-departure events (specifically the reaction time exceeded three times the alert reaction time). The results of our previous studies revealed that arousing feedback immediately reversed deteriorating driving performance, which was accompanied by concurrent EEG theta- and alpha-power suppression in the bilateral occipital areas. This study further proposes a feedback efficacy assessment system to accurately estimate the efficacy of arousing warning signals delivered to drowsy participants by monitoring the changes in their EEG power spectra immediately thereafter. The classification accuracy was up 77.8% for determining the need for triggering additional warning signals. The findings of this study, in conjunction with previous studies on EEG correlates of behavioral lapses, might lead to a practical closed-loop system to predict, monitor and rectify behavioral lapses of human operators in attention-critical settings.

  3. Quantitative analysis on collagen of dermatofibrosarcoma protuberans skin by second harmonic generation microscopy.

    PubMed

    Wu, Shulian; Huang, Yudian; Li, Hui; Wang, Yunxia; Zhang, Xiaoman

    2015-01-01

    Dermatofibrosarcoma protuberans (DFSP) is a skin cancer usually mistaken as other benign tumors. Abnormal DFSP resection results in tumor recurrence. Quantitative characterization of collagen alteration on the skin tumor is essential for developing a diagnostic technique. In this study, second harmonic generation (SHG) microscopy was performed to obtain images of the human DFSP skin and normal skin. Subsequently, structure and texture analysis methods were applied to determine the differences in skin texture characteristics between the two skin types, and the link between collagen alteration and tumor was established. Results suggest that combining SHG microscopy and texture analysis methods is a feasible and effective method to describe the characteristics of skin tumor like DFSP. © Wiley Periodicals, Inc.

  4. Real-time fluorescence microscopy monitoring of porphyrin biodistribution

    NASA Astrophysics Data System (ADS)

    Kimel, Sol; Gottfried, Varda; Kunzi-Rapp, Karin; Akguen, Nermin; Schneckenburger, Herbert

    1996-01-01

    In vivo uptake of the natural porphyrins, uroporphyrin III (UP), coproporphyrin III (CP) and protoporphyrin IX (PP), was monitored by fluorescence microscopy. Experiments were performed using the chick chorioallantoic membrane (CAM) model, which allowed video documentation of fluorescence both in real time and after integration over a chosen time interval (usually 2 s). Sensitizers at a concentration of 50 (mu) M (100 (mu) L) were injected into a medium-sized vein (diameter approximately 40 micrometer) using an ultra-fine 10 micrometer diameter needle. Fluorescence images were quantitated by subtracting the fluorescence intensity of surrounding CAM tissue (Fmatrix) from the intravascular fluorescence intensity (Fintravascular), after transformation of the video frames into digital form. The differential fluorescence intensity, Fintravascular - Fmatrix, is a measure of the biodistribution. Real time measurements clearly showed that CP and UP fluorescence is associated with moving erythrocytes and not with endothelial cells of the vessel wall. Fluorescence intensity was monitored, up to 60 minutes after injection, by averaging the fluorescence over time intervals of 2 s and recording the integrated images. The fluorescence intensity reached its maximum in about 20 - 30 min after injection, presumably after monomerization inside erythrocyte membranes. The results are interpreted in terms of physical-chemical characteristics (e.g. hydrophilicity) and correlated with the photodynamically induced hemostasis in CAM blood vessels.

  5. Time-lapse imaging of human heart motion with switched array UWB radar.

    PubMed

    Brovoll, Sverre; Berger, Tor; Paichard, Yoann; Aardal, Øyvind; Lande, Tor Sverre; Hamran, Svein-Erik

    2014-10-01

    Radar systems for detection of human heartbeats have mostly been single-channel systems with limited spatial resolution. In this paper, a radar system for ultra-wideband (UWB) imaging of the human heart is presented. To make the radar waves penetrate the human tissue the antenna is placed very close to the body. The antenna is an array with eight elements, and an antenna switch system connects the radar to the individual elements in sequence to form an image. Successive images are used to build up time-lapse movies of the beating heart. Measurements on a human test subject are presented and the heart motion is estimated at different locations inside the body. The movies show rhythmic motion consistent with the beating heart, and the location and shape of the reflections correspond well with the expected response form the heart wall. The spatial dependent heart motion is compared to ECG recordings, and it is confirmed that heartbeat modulations are seen in the radar data. This work shows that radar imaging of the human heart may provide valuable information on the mechanical movement of the heart.

  6. Real-time digital signal processing in multiphoton and time-resolved microscopy

    NASA Astrophysics Data System (ADS)

    Wilson, Jesse W.; Warren, Warren S.; Fischer, Martin C.

    2016-03-01

    The use of multiphoton interactions in biological tissue for imaging contrast requires highly sensitive optical measurements. These often involve signal processing and filtering steps between the photodetector and the data acquisition device, such as photon counting and lock-in amplification. These steps can be implemented as real-time digital signal processing (DSP) elements on field-programmable gate array (FPGA) devices, an approach that affords much greater flexibility than commercial photon counting or lock-in devices. We will present progress toward developing two new FPGA-based DSP devices for multiphoton and time-resolved microscopy applications. The first is a high-speed multiharmonic lock-in amplifier for transient absorption microscopy, which is being developed for real-time analysis of the intensity-dependence of melanin, with applications in vivo and ex vivo (noninvasive histopathology of melanoma and pigmented lesions). The second device is a kHz lock-in amplifier running on a low cost (50-200) development platform. It is our hope that these FPGA-based DSP devices will enable new, high-speed, low-cost applications in multiphoton and time-resolved microscopy.

  7. Nanoscale nuclear architecture for cancer diagnosis beyond pathology via spatial-domain low-coherence quantitative phase microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Pin; Bista, Rajan K.; Khalbuss, Walid E.; Qiu, Wei; Uttam, Shikhar; Staton, Kevin; Zhang, Lin; Brentnall, Teresa A.; Brand, Randall E.; Liu, Yang

    2010-11-01

    Definitive diagnosis of malignancy is often challenging due to limited availability of human cell or tissue samples and morphological similarity with certain benign conditions. Our recently developed novel technology-spatial-domain low-coherence quantitative phase microscopy (SL-QPM)-overcomes the technical difficulties and enables us to obtain quantitative information about cell nuclear architectural characteristics with nanoscale sensitivity. We explore its ability to improve the identification of malignancy, especially in cytopathologically non-cancerous-appearing cells. We perform proof-of-concept experiments with an animal model of colorectal carcinogenesis-APCMin mouse model and human cytology specimens of colorectal cancer. We show the ability of in situ nanoscale nuclear architectural characteristics in identifying cancerous cells, especially in those labeled as ``indeterminate or normal'' by expert cytopathologists. Our approach is based on the quantitative analysis of the cell nucleus on the original cytology slides without additional processing, which can be readily applied in a conventional clinical setting. Our simple and practical optical microscopy technique may lead to the development of novel methods for early detection of cancer.

  8. Exploring the interactome: microfluidic isolation of proteins and interacting partners for quantitative analysis by electron microscopy.

    PubMed

    Giss, Dominic; Kemmerling, Simon; Dandey, Venkata; Stahlberg, Henning; Braun, Thomas

    2014-05-20

    Multimolecular protein complexes are important for many cellular processes. However, the stochastic nature of the cellular interactome makes the experimental detection of complex protein assemblies difficult and quantitative analysis at the single molecule level essential. Here, we present a fast and simple microfluidic method for (i) the quantitative isolation of endogenous levels of untagged protein complexes from minute volumes of cell lysates under close to physiological conditions and (ii) the labeling of specific components constituting these complexes. The method presented uses specific antibodies that are conjugated via a photocleavable linker to magnetic beads that are trapped in microcapillaries to immobilize the target proteins. Proteins are released by photocleavage, eluted, and subsequently analyzed by quantitative transmission electron microscopy at the single molecule level. Additionally, before photocleavage, immunogold can be employed to label proteins that interact with the primary target protein. Thus, the presented method provides a new way to study the interactome and, in combination with single molecule transmission electron microscopy, to structurally characterize the large, dynamic, heterogeneous multimolecular protein complexes formed.

  9. 4D inversion of time-lapse magnetotelluric data sets for monitoring geothermal reservoir

    NASA Astrophysics Data System (ADS)

    Nam, Myung Jin; Song, Yoonho; Jang, Hannuree; Kim, Bitnarae

    2017-06-01

    The productivity of a geothermal reservoir, which is a function of the pore-space and fluid-flow path of the reservoir, varies since the properties of the reservoir changes with geothermal reservoir production. Because the variation in the reservoir properties causes changes in electrical resistivity, time-lapse (TL) three-dimensional (3D) magnetotelluric (MT) methods can be applied to monitor the productivity variation of a geothermal reservoir thanks to not only its sensitivity to the electrical resistivity but also its deep depth of survey penetration. For an accurate interpretation of TL MT-data sets, a four-dimensional (4D) MT inversion algorithm has been developed to simultaneously invert all vintage data considering time-coupling between vintages. However, the changes in electrical resistivity of deep geothermal reservoirs are usually small generating minimum variation in TL MT responses. Maximizing the sensitivity of inversion to the changes in resistivity is critical in the success of 4D MT inversion. Thus, we further developed a focused 4D MT inversion method by considering not only the location of a reservoir but also the distribution of newly-generated fractures during the production. For the evaluation of the 4D MT algorithm, we tested our 4D inversion algorithms using synthetic TL MT-data sets.

  10. Respiratory problems and anxiety sensitivity in smoking lapse among treatment seeking smokers.

    PubMed

    Zvolensky, Michael J; Rodríguez-Cano, Rubén; Paulus, Daniel J; Kotov, Roman; Bromet, Evelyn; Gonzalez, Adam; Manning, Kara; Luft, Benjamin J

    2017-12-01

    The current study examined whether the interaction of lower respiratory symptoms and anxiety sensitivity is related to smoking lapse in the context of smoking cessation. Participants were adult daily smokers (N=60) exposed to the World Trade Center (WTC) disaster who were in a smoking cessation treatment program (75.0% male, 50.6years old [SD=9.2], and current smoking rate was 17.6 cigarettes per day (SD=10.6). Results indicated that the interaction between lower respiratory symptoms and anxiety sensitivity was a significant predictor of greater risk for lapse (i.e., lower survival time; B=0.005, OR=1.01, p=0.039). Follow-up analysis showed that greater respiratory symptoms were a significant predictor of lapse risk among those with high (B=0.116, OR=1.12, p=0.025), but not those with low (B=-0.048, OR=0.95, p=0.322), levels of anxiety sensitivity. The findings from the current study suggest that smokers with greater respiratory symptoms and higher levels of anxiety sensitivity may be associated with early lapse to smoking following smoking cessation treatment. Future work has the potential to inform the development of tailored cessation interventions for smokers who experience varying levels of lower respiratory symptoms and anxiety sensitivity. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Quantitative Near-field Microscopy of Heterogeneous and Correlated Electron Oxides

    NASA Astrophysics Data System (ADS)

    McLeod, Alexander Swinton

    Scanning near-field optical microscopy (SNOM) is a novel scanning probe microscopy technique capable of circumventing the conventional diffraction limit of light, affording unparalleled optical resolution (down to 10 nanometers) even for radiation in the infrared and terahertz energy regimes, with light wavelengths exceeding 10 micrometers. However, although this technique has been developed and employed for more than a decade to a qualitatively impressive effect, researchers have lacked a practically quantitative grasp of its capabilities, and its application scope has so far remained restricted by implementations limited to ambient atmospheric conditions. The two-fold objective of this dissertation work has been to address both these shortcomings. The first half of the dissertation presents a realistic, semi-analytic, and benchmarked theoretical description of probe-sample near-field interactions that form the basis of SNOM. Owing its name to the efficient nano-focusing of light at a sharp metallic apex, the "lightning rod model" of probe-sample near-field interactions is mathematically developed from a flexible and realistic scattering formalism. Powerful and practical applications are demonstrated through the accurate prediction of spectroscopic near-field optical contrasts, as well as the "inversion" of these spectroscopic contrasts into a quantitative description of material optical properties. Thus enabled, this thesis work proceeds to present quantitative applications of infrared near-field spectroscopy to investigate nano-resolved chemical compositions in a diverse host of samples, including technologically relevant lithium ion battery materials, astrophysical planetary materials, and invaluable returned extraterrestrial samples. The second half of the dissertation presents the design, construction, and demonstration of a sophisticated low-temperature scanning near-field infrared microscope. This instrument operates in an ultra-high vacuum environment

  12. Intracellular subsurface imaging using a hybrid shear-force feedback/scanning quantitative phase microscopy technique

    NASA Astrophysics Data System (ADS)

    Edward, Kert

    Quantitative phase microscopy (QPM) allows for the imaging of translucent or transparent biological specimens without the need for exogenous contrast agents. This technique is usually applied towards the investigation of simple cells such as red blood cells which are typically enucleated and can be considered to be homogenous. However, most biological cells are nucleated and contain other interesting intracellular organelles. It has been established that the physical characteristics of certain subsurface structures such as the shape and roughness of the nucleus is well correlated with onset and progress of pathological conditions such as cancer. Although the acquired quantitative phase information of biological cells contains surface information as well as coupled subsurface information, the latter has been ignored up until now. A novel scanning quantitative phase imaging system unencumbered by 2pi ambiguities is hereby presented. This system is incorporated into a shear-force feedback scheme which allows for simultaneous phase and topography determination. It will be shown how subsequent image processing of these two data sets allows for the extraction of the subsurface component in the phase data and in vivo cell refractometry studies. Both fabricated samples and biological cells ranging from rat fibroblast cells to malaria infected human erythrocytes were investigated as part of this research. The results correlate quite well with that obtained via other microscopy techniques.

  13. Toward quantitative estimation of material properties with dynamic mode atomic force microscopy: a comparative study.

    PubMed

    Ghosal, Sayan; Gannepalli, Anil; Salapaka, Murti

    2017-08-11

    In this article, we explore methods that enable estimation of material properties with the dynamic mode atomic force microscopy suitable for soft matter investigation. The article presents the viewpoint of casting the system, comprising of a flexure probe interacting with the sample, as an equivalent cantilever system and compares a steady-state analysis based method with a recursive estimation technique for determining the parameters of the equivalent cantilever system in real time. The steady-state analysis of the equivalent cantilever model, which has been implicitly assumed in studies on material property determination, is validated analytically and experimentally. We show that the steady-state based technique yields results that quantitatively agree with the recursive method in the domain of its validity. The steady-state technique is considerably simpler to implement, however, slower compared to the recursive technique. The parameters of the equivalent system are utilized to interpret storage and dissipative properties of the sample. Finally, the article identifies key pitfalls that need to be avoided toward the quantitative estimation of material properties.

  14. Systematic characterization of maturation time of fluorescent proteins in living cells

    PubMed Central

    Balleza, Enrique; Kim, J. Mark; Cluzel, Philippe

    2017-01-01

    Slow maturation time of fluorescent proteins limits accurate measurement of rapid gene expression dynamics and effectively reduces fluorescence signal in growing cells. We used high-precision time-lapse microscopy to characterize, at two different temperatures in E. coli, the maturation kinetics of 50 FPs that span the visible spectrum. We identified fast-maturing FPs that yield the highest signal-to-noise ratio and temporal resolution in individual growing cells. PMID:29320486

  15. Quantitative analyses of the 3D nuclear landscape recorded with super-resolved fluorescence microscopy.

    PubMed

    Schmid, Volker J; Cremer, Marion; Cremer, Thomas

    2017-07-01

    Recent advancements of super-resolved fluorescence microscopy have revolutionized microscopic studies of cells, including the exceedingly complex structural organization of cell nuclei in space and time. In this paper we describe and discuss tools for (semi-) automated, quantitative 3D analyses of the spatial nuclear organization. These tools allow the quantitative assessment of highly resolved different chromatin compaction levels in individual cell nuclei, which reflect functionally different regions or sub-compartments of the 3D nuclear landscape, and measurements of absolute distances between sites of different chromatin compaction. In addition, these tools allow 3D mapping of specific DNA/RNA sequences and nuclear proteins relative to the 3D chromatin compaction maps and comparisons of multiple cell nuclei. The tools are available in the free and open source R packages nucim and bioimagetools. We discuss the use of masks for the segmentation of nuclei and the use of DNA stains, such as DAPI, as a proxy for local differences in chromatin compaction. We further discuss the limitations of 3D maps of the nuclear landscape as well as problems of the biological interpretation of such data. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders.

    PubMed

    Marquet, Pierre; Depeursinge, Christian; Magistretti, Pierre J

    2014-10-01

    Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

  17. Review of quantitative phase-digital holographic microscopy: promising novel imaging technique to resolve neuronal network activity and identify cellular biomarkers of psychiatric disorders

    PubMed Central

    Marquet, Pierre; Depeursinge, Christian; Magistretti, Pierre J.

    2014-01-01

    Abstract. Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed. PMID:26157976

  18. Imaging and quantitative data acquisition of biological cell walls with Atomic Force Microscopy and Scanning Acoustic Microscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tittmann, B. R.; Xi, X.

    This chapter demonstrates the feasibility of Atomic Force Microscopy (AFM) and High Frequency Scanning Acoustic Microscopy (HF-SAM) as tools to characterize biological tissues. Both the AFM and the SAM have shown to provide imaging (with different resolution) and quantitative elasticity measuring abilities. Plant cell walls with minimal disturbance and under conditions of their native state have been examined with these two kinds of microscopy. After descriptions of both the SAM and AFM, their special features and the typical sample preparation is discussed. The sample preparation is focused here on epidermal peels of onion scales and celery epidermis cells which weremore » sectioned for the AFM to visualize the inner surface (closest to the plasma membrane) of the outer epidermal wall. The nm-wide cellulose microfibrils orientation and multilayer structure were clearly observed. The microfibril orientation and alignment tend to be more organized in older scales compared with younger scales. The onion epidermis cell wall was also used as a test analog to study cell wall elasticity by the AFM nanoindentation and the SAM V(z) feature. The novelty in this work was to demonstrate the capability of these two techniques to analyze isolated, single layered plant cell walls in their natural state. AFM nanoindentation was also used to probe the effects of Ethylenediaminetetraacetic acid (EDTA), and calcium ion treatment to modify pectin networks in cell walls. The results suggest a significant modulus increase in the calcium ion treatment and a slight decrease in EDTA treatment. To complement the AFM measurements, the HF-SAM was used to obtain the V(z) signatures of the onion epidermis. These measurements were focused on documenting the effect of pectinase enzyme treatment. The results indicate a significant change in the V(z) signature curves with time into the enzyme treatment. Thus AFM and HF-SAM open the door to a systematic nondestructive structure and mechanical

  19. Time-lapse Mise-á-la-Masse measurements and modeling for tracer test monitoring in a shallow aquifer

    NASA Astrophysics Data System (ADS)

    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.

  20. Dispositional Mindfulness Predicts Enhanced Smoking Cessation and Smoking Lapse Recovery

    PubMed Central

    Heppner, Whitney L.; Spears, Claire Adams; Correa-Fernández, Virmarie; Castro, Yessenia; Li, Yisheng; Guo, Beibei; Reitzel, Lorraine R.; Vidrine, Jennifer Irvin; Mazas, Carlos A.; Cofta-Woerpel, Ludmila; Cinciripini, Paul M.; Ahluwalia, Jasjit S.; Wetter, David W.

    2016-01-01

    Background Although mindfulness has been hypothesized to promote health behaviors, no research has examined how dispositional mindfulness might influence the process of smoking cessation. Purpose The current study investigated dispositional mindfulness, smoking abstinence, and recovery from a lapse among African American smokers. Methods Participants were 399 African Americans seeking smoking cessation treatment (treatments did not include any components related to mindfulness). Dispositional mindfulness and other psychosocial measures were obtained pre-quit; smoking abstinence was assessed 3 days, 31 days, and 26 weeks post-quit. Results Individuals higher in dispositional mindfulness were more likely to quit smoking both initially and over time. Moreover, among individuals who had lapsed at day 3, those higher in mindfulness were more likely to recover abstinence by the later time points. The mindfulness-early abstinence association was mediated by lower negative affect, lower expectancies to regulate affect via smoking, and higher perceived social support. Conclusions Results suggest that mindfulness might enhance smoking cessation among African American smokers by operating on mechanisms posited by prominent models of addiction. PMID:26743533

  1. Live imaging of muscles in Drosophila metamorphosis: Towards high-throughput gene identification and function analysis.

    PubMed

    Puah, Wee Choo; Wasser, Martin

    2016-03-01

    Time-lapse microscopy in developmental biology is an emerging tool for functional genomics. Phenotypic effects of gene perturbations can be studied non-invasively at multiple time points in chronological order. During metamorphosis of Drosophila melanogaster, time-lapse microscopy using fluorescent reporters allows visualization of alternative fates of larval muscles, which are a model for the study of genes related to muscle wasting. While doomed muscles enter hormone-induced programmed cell death, a smaller population of persistent muscles survives to adulthood and undergoes morphological remodeling that involves atrophy in early, and hypertrophy in late pupation. We developed a method that combines in vivo imaging, targeted gene perturbation and image analysis to identify and characterize genes involved in muscle development. Macrozoom microscopy helps to screen for interesting muscle phenotypes, while confocal microscopy in multiple locations over 4-5 days produces time-lapse images that are used to quantify changes in cell morphology. Performing a similar investigation using fixed pupal tissues would be too time-consuming and therefore impractical. We describe three applications of our pipeline. First, we show how quantitative microscopy can track and measure morphological changes of muscle throughout metamorphosis and analyze genes involved in atrophy. Second, our assay can help to identify genes that either promote or prevent histolysis of abdominal muscles. Third, we apply our approach to test new fluorescent proteins as live markers for muscle development. We describe mKO2 tagged Cysteine proteinase 1 (Cp1) and Troponin-I (TnI) as examples of proteins showing developmental changes in subcellular localization. Finally, we discuss strategies to improve throughput of our pipeline to permit genome-wide screens in the future. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  2. Quantitative measurements of nanoscale permittivity and conductivity using tuning-fork-based microwave impedance microscopy

    NASA Astrophysics Data System (ADS)

    Wu, Xiaoyu; Hao, Zhenqi; Wu, Di; Zheng, Lu; Jiang, Zhanzhi; Ganesan, Vishal; Wang, Yayu; Lai, Keji

    2018-04-01

    We report quantitative measurements of nanoscale permittivity and conductivity using tuning-fork (TF) based microwave impedance microscopy (MIM). The system is operated under the driving amplitude modulation mode, which ensures satisfactory feedback stability on samples with rough surfaces. The demodulated MIM signals on a series of bulk dielectrics are in good agreement with results simulated by finite-element analysis. Using the TF-MIM, we have visualized the evolution of nanoscale conductance on back-gated MoS2 field effect transistors, and the results are consistent with the transport data. Our work suggests that quantitative analysis of mesoscopic electrical properties can be achieved by near-field microwave imaging with small distance modulation.

  3. Time-Lapse Cinemicrographic Studies of X-Irradiated HeLa S3 Cells

    PubMed Central

    Hurwitz, Camilla; Tolmach, L. J.

    1969-01-01

    Analysis of time-lapse cinemicrographs of X-irradiated HeLa S3 cells has shown that the incidence of cell fusion was increased from 0.9% (following 1267 divisions) in control cells to an average of 22% (following 655 divisions) in cells irradiated with 500 rad doses of 220 kv X-rays. The incidence depended on the stage of the generation cycle at which the parent cells were irradiated. It was nearly constant in the first three postirradiation generations. Fusion occurred at all stages of the generation cycle, but preferentially during the first 20%. Cells undergoing fusion progressed more slowly through the generation cycle and had a higher probability of disintegrating than did irradiated cells that did not fuse. The occurrence of fusion was clonally distributed in the population. It took place only between sister (or closely related) cells. Protoplasmic bridges were often visible between sister cells prior to fusion. Giant cells arose only as a result of fusion. The incidence of multipolar divisions, though higher than in unirradiated cells, was only 5.5% in cultures irradiated with 500 rads. Fusion occurred following 85% of the multipolar divisions and was often followed by a multipolar division. ImagesFigure 1 PMID:5807221

  4. Difficulties with Emotion Regulation and Psychopathology Interact to Predict Early Smoking Cessation Lapse

    PubMed Central

    Zvolensky, Michael J.; Schmidt, Norman B.

    2015-01-01

    There is little knowledge about how emotion regulation difficulties interplay with psychopathology in terms of smoking cessation. Participants (n = 250; 53.2 % female, Mage = 39.5, SD = 13.85) were community-recruited daily smokers (≥8 cigarettes per day) who self-reported motivation to quit smoking; 38.8 % of the sample met criteria for a current (past 12-month) psychological disorder. Emotion regulation deficits were assessed pre-quit using the Difficulties with Emotion Regulation Scale (DERS; Gratz and Roemer in J Psychopathol Behav Assess 26(1):41–54, 2004) and smoking behavior in the 28 days post-quit was assessed using the Timeline Follow-Back (TLFB; Sobell and Sobell in Measuring alcohol consumption: psychosocial and biochemical methods. Humana Press, Totowa, 1992). A Cox proportional-hazard regression analysis was used to model the effects of past-year psychopathology, DERS (total score), and their interaction, in terms of time to lapse post-quit day. After adjusting for the effects of gender, age, pre-quit level of nicotine dependence, and treatment condition, the model revealed a non-significant effect of past-year psychopathology (OR = 1.14, CI95 % = 0.82–1.61) and difficulties with emotion regulation (OR = 1.01, CI95 % = 1.00–1.01) on likelihood of lapse rate. However, the interactive effect of psychopathology status and difficulties with emotion regulation was significant (OR = 0.98, CI95 % = 0.97–0.99). Specifically, there was a significant conditional effect of psychopathology status on lapse rate likelihood at low, but not high, levels of emotion regulation difficulties. Plots of the cumulative survival functions indicated that for smokers without a past-year psychological disorder, those with lower DERS scores relative to elevated DERS scores had significantly lower likelihood of early smoking lapse, whereas for smokers with past-year psychopathology, DERS scores did not differentially impact lapse rate likelihood. Smokers with emotion

  5. Difficulties with Emotion Regulation and Psychopathology Interact to Predict Early Smoking Cessation Lapse.

    PubMed

    Farris, Samantha G; Zvolensky, Michael J; Schmidt, Norman B

    2016-06-01

    There is little knowledge about how emotion regulation difficulties interplay with psychopathology in terms of smoking cessation. Participants ( n = 250; 53.2 % female, M age = 39.5, SD = 13.85) were community-recruited daily smokers (≥8 cigarettes per day) who self-reported motivation to quit smoking; 38.8 % of the sample met criteria for a current (past 12-month) psychological disorder. Emotion regulation deficits were assessed pre-quit using the Difficulties with Emotion Regulation Scale (DERS; Gratz and Roemer in J Psychopathol Behav Assess 26(1):41-54, 2004) and smoking behavior in the 28 days post-quit was assessed using the Timeline Follow-Back (TLFB; Sobell and Sobell in Measuring alcohol consumption: psychosocial and biochemical methods. Humana Press, Totowa, 1992). A Cox proportional-hazard regression analysis was used to model the effects of past-year psychopathology, DERS (total score), and their interaction, in terms of time to lapse post-quit day. After adjusting for the effects of gender, age, pre-quit level of nicotine dependence, and treatment condition, the model revealed a non-significant effect of past-year psychopathology ( OR = 1.14, CI 95 % = 0.82-1.61) and difficulties with emotion regulation ( OR = 1.01, CI 95 % = 1.00-1.01) on likelihood of lapse rate. However, the interactive effect of psychopathology status and difficulties with emotion regulation was significant ( OR = 0.98, CI 95 % = 0.97-0.99). Specifically, there was a significant conditional effect of psychopathology status on lapse rate likelihood at low, but not high, levels of emotion regulation difficulties. Plots of the cumulative survival functions indicated that for smokers without a past-year psychological disorder, those with lower DERS scores relative to elevated DERS scores had significantly lower likelihood of early smoking lapse, whereas for smokers with past-year psychopathology, DERS scores did not differentially impact lapse rate likelihood. Smokers with emotion

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

    PubMed

    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

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

    PubMed Central

    2014-01-01

    Background 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. Methods 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. Results 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

  8. Cell Motility Dynamics: A Novel Segmentation Algorithm to Quantify Multi-Cellular Bright Field Microscopy Images

    PubMed Central

    Zaritsky, Assaf; Natan, Sari; Horev, Judith; Hecht, Inbal; Wolf, Lior; Ben-Jacob, Eshel; Tsarfaty, Ilan

    2011-01-01

    Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs) is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF) on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC) images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional fluorescence single

  9. Cell motility dynamics: a novel segmentation algorithm to quantify multi-cellular bright field microscopy images.

    PubMed

    Zaritsky, Assaf; Natan, Sari; Horev, Judith; Hecht, Inbal; Wolf, Lior; Ben-Jacob, Eshel; Tsarfaty, Ilan

    2011-01-01

    Confocal microscopy analysis of fluorescence and morphology is becoming the standard tool in cell biology and molecular imaging. Accurate quantification algorithms are required to enhance the understanding of different biological phenomena. We present a novel approach based on image-segmentation of multi-cellular regions in bright field images demonstrating enhanced quantitative analyses and better understanding of cell motility. We present MultiCellSeg, a segmentation algorithm to separate between multi-cellular and background regions for bright field images, which is based on classification of local patches within an image: a cascade of Support Vector Machines (SVMs) is applied using basic image features. Post processing includes additional classification and graph-cut segmentation to reclassify erroneous regions and refine the segmentation. This approach leads to a parameter-free and robust algorithm. Comparison to an alternative algorithm on wound healing assay images demonstrates its superiority. The proposed approach was used to evaluate common cell migration models such as wound healing and scatter assay. It was applied to quantify the acceleration effect of Hepatocyte growth factor/scatter factor (HGF/SF) on healing rate in a time lapse confocal microscopy wound healing assay and demonstrated that the healing rate is linear in both treated and untreated cells, and that HGF/SF accelerates the healing rate by approximately two-fold. A novel fully automated, accurate, zero-parameters method to classify and score scatter-assay images was developed and demonstrated that multi-cellular texture is an excellent descriptor to measure HGF/SF-induced cell scattering. We show that exploitation of textural information from differential interference contrast (DIC) images on the multi-cellular level can prove beneficial for the analyses of wound healing and scatter assays. The proposed approach is generic and can be used alone or alongside traditional fluorescence single

  10. Application of RVA and Time-Lapse Photography to Explore Effects of Extent of Chlorination, Milling Extraction Rate, and Particle-Size Reduction of Flour on Cake-Baking Functionality

    USDA-ARS?s Scientific Manuscript database

    Three factors (extent of chlorination, milling extraction rate and particle-size reduction) in the cake-bakeing functionality of Croplan 594W flour were explored by Rapid Visco-Analyzer (RVA) and time-lapse photography. The extent of chlorination and milling extraction rate showed dramatic effects,...

  11. New Tools for Comparing Microscopy Images: Quantitative Analysis of Cell Types in Bacillus subtilis

    PubMed Central

    van Gestel, Jordi; Vlamakis, Hera

    2014-01-01

    Fluorescence microscopy is a method commonly used to examine individual differences between bacterial cells, yet many studies still lack a quantitative analysis of fluorescence microscopy data. Here we introduce some simple tools that microbiologists can use to analyze and compare their microscopy images. We show how image data can be converted to distribution data. These data can be subjected to a cluster analysis that makes it possible to objectively compare microscopy images. The distribution data can further be analyzed using distribution fitting. We illustrate our methods by scrutinizing two independently acquired data sets, each containing microscopy images of a doubly labeled Bacillus subtilis strain. For the first data set, we examined the expression of srfA and tapA, two genes which are expressed in surfactin-producing and matrix-producing cells, respectively. For the second data set, we examined the expression of eps and tapA; these genes are expressed in matrix-producing cells. We show that srfA is expressed by all cells in the population, a finding which contrasts with a previously reported bimodal distribution of srfA expression. In addition, we show that eps and tapA do not always have the same expression profiles, despite being expressed in the same cell type: both operons are expressed in cell chains, while single cells mainly express eps. These findings exemplify that the quantification and comparison of microscopy data can yield insights that otherwise would go unnoticed. PMID:25448819

  12. Quantitative study of mammalian cells by scanning transmission soft X-ray microscopy

    NASA Astrophysics Data System (ADS)

    Shinohara, K.; Ohigashi, T.; Toné, S.; Kado, M.; Ito, A.

    2017-06-01

    Molecular distribution in mammalian cells was studied by soft X-ray scanning transmission microscopy with respect to the quantitative aspect of analysis. NEXAFS profiles at the C, N and O K-absorption edges were combined and used for the analysis. For the estimation of quantity for nucleic acids and proteins, NEXAFS profiles of DNA and bovine serum albumin (BSA) at the N K-absorption edge were applied assuming that those were their representatives. The method has a potential to explore the other molecular components than nucleic acids and proteins.

  13. Monitoring of In-Situ Remediation By Time Lapse 3D Geo-Electric Measurements

    NASA Astrophysics Data System (ADS)

    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.

  14. Real-time high dynamic range laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

    2016-04-01

    In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging.

  15. Creation of a bovine herpes virus 1 (BoHV-1) quantitative particle standard by transmission electron microscopy and comparison with established standards for use in real-time PCR.

    PubMed

    Hoferer, Marc; Braun, Anne; Sting, Reinhard

    2017-07-01

    Standards are pivotal for pathogen quantification by real-time PCR (qPCR); however, the creation of a complete and universally applicable virus particle standard is challenging. In the present study a procedure based on purification of bovine herpes virus type 1 (BoHV-1) and subsequent quantification by transmission electron microscopy (TEM) is described. Accompanying quantitative quality controls of the TEM preparation procedure using qPCR yielded recovery rates of more than 95% of the BoHV-1 virus particles on the grid used for virus counting, which was attributed to pre-treatment of the grid with 5% bovine albumin. To compare the value of the new virus particle standard for use in qPCR, virus counter based quantification and established pure DNA standards represented by a plasmid and an oligonucleotide were included. It could be shown that the numbers of virus particles, plasmid and oligonucleotide equivalents were within one log10 range determined on the basis of standard curves indicating that different approaches provide comparable quantitative values. However, only virus particles represent a complete, universally applicable quantitative virus standard that meets the high requirements of an RNA and DNA virus gold standard. In contrast, standards based on pure DNA have to be considered as sub-standard due to limited applications. Copyright © 2017 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  16. Tracking neutrophil intraluminal crawling, transendothelial migration and chemotaxis in tissue by intravital video microscopy.

    PubMed

    Xu, Najia; Lei, Xi; Liu, Lixin

    2011-09-24

    The recruitment of circulating leukocytes from blood stream to the inflamed tissue is a crucial and complex process of inflammation(1,2). In the postcapillary venules of inflamed tissue, leukocytes initially tether and roll on the luminal surface of venular wall. Rolling leukocytes arrest on endothelium and undergo firm adhesion in response to chemokine or other chemoattractants on the venular surface. Many adherent leukocytes relocate from the initial site of adhesion to the junctional extravasation site in endothelium, a process termed intraluminal crawling(3). Following crawling, leukocytes move across endothelium (transmigration) and migrate in extravascular tissue toward the source of chemoattractant (chemotaxis)(4). Intravital microscopy is a powerful tool for visualizing leukocyte-endothelial cell interactions in vivo and revealing cellular and molecular mechanisms of leukocyte recruitment(2,5). In this report, we provide a comprehensive description of using brightfield intravital microscopy to visualize and determine the detailed processes of neutrophil recruitment in mouse cremaster muscle in response to the gradient of a neutrophil chemoattractant. To induce neutrophil recruitment, a small piece of agarose gel (~1-mm(3) size) containing neutrophil chemoattractant MIP-2 (CXCL2, a CXC chemokine) or WKYMVm (Trp-Lys-Tyr-Val-D-Met, a synthetic analog of bacterial peptide) is placed on the muscle tissue adjacent to the observed postcapillary venule. With time-lapsed video photography and computer software ImageJ, neutrophil intraluminal crawling on endothelium, neutrophil transendothelial migration and the migration and chemotaxis in tissue are visualized and tracked. This protocol allows reliable and quantitative analysis of many neutrophil recruitment parameters such as intraluminal crawling velocity, transmigration time, detachment time, migration velocity, chemotaxis velocity and chemotaxis index in tissue. We demonstrate that using this protocol, these

  17. Mapping surface charge density of lipid bilayers by quantitative surface conductivity microscopy

    PubMed Central

    Klausen, Lasse Hyldgaard; Fuhs, Thomas; Dong, Mingdong

    2016-01-01

    Local surface charge density of lipid membranes influences membrane–protein interactions leading to distinct functions in all living cells, and it is a vital parameter in understanding membrane-binding mechanisms, liposome design and drug delivery. Despite the significance, no method has so far been capable of mapping surface charge densities under physiologically relevant conditions. Here, we use a scanning nanopipette setup (scanning ion-conductance microscope) combined with a novel algorithm to investigate the surface conductivity near supported lipid bilayers, and we present a new approach, quantitative surface conductivity microscopy (QSCM), capable of mapping surface charge density with high-quantitative precision and nanoscale resolution. The method is validated through an extensive theoretical analysis of the ionic current at the nanopipette tip, and we demonstrate the capacity of QSCM by mapping the surface charge density of model cationic, anionic and zwitterionic lipids with results accurately matching theoretical values. PMID:27561322

  18. Mapping surface charge density of lipid bilayers by quantitative surface conductivity microscopy

    NASA Astrophysics Data System (ADS)

    Klausen, Lasse Hyldgaard; Fuhs, Thomas; Dong, Mingdong

    2016-08-01

    Local surface charge density of lipid membranes influences membrane-protein interactions leading to distinct functions in all living cells, and it is a vital parameter in understanding membrane-binding mechanisms, liposome design and drug delivery. Despite the significance, no method has so far been capable of mapping surface charge densities under physiologically relevant conditions. Here, we use a scanning nanopipette setup (scanning ion-conductance microscope) combined with a novel algorithm to investigate the surface conductivity near supported lipid bilayers, and we present a new approach, quantitative surface conductivity microscopy (QSCM), capable of mapping surface charge density with high-quantitative precision and nanoscale resolution. The method is validated through an extensive theoretical analysis of the ionic current at the nanopipette tip, and we demonstrate the capacity of QSCM by mapping the surface charge density of model cationic, anionic and zwitterionic lipids with results accurately matching theoretical values.

  19. Mapping surface charge density of lipid bilayers by quantitative surface conductivity microscopy.

    PubMed

    Klausen, Lasse Hyldgaard; Fuhs, Thomas; Dong, Mingdong

    2016-08-26

    Local surface charge density of lipid membranes influences membrane-protein interactions leading to distinct functions in all living cells, and it is a vital parameter in understanding membrane-binding mechanisms, liposome design and drug delivery. Despite the significance, no method has so far been capable of mapping surface charge densities under physiologically relevant conditions. Here, we use a scanning nanopipette setup (scanning ion-conductance microscope) combined with a novel algorithm to investigate the surface conductivity near supported lipid bilayers, and we present a new approach, quantitative surface conductivity microscopy (QSCM), capable of mapping surface charge density with high-quantitative precision and nanoscale resolution. The method is validated through an extensive theoretical analysis of the ionic current at the nanopipette tip, and we demonstrate the capacity of QSCM by mapping the surface charge density of model cationic, anionic and zwitterionic lipids with results accurately matching theoretical values.

  20. 40 CFR 60.2655 - How do I renew my lapsed operator qualification?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Emissions Guidelines and Compliance Times for Commercial and Industrial Solid Waste Incineration Units Model Rule-Operator Training...) For a lapse of less than 3 years, you must complete a standard annual refresher course described in...

  1. 40 CFR 60.2655 - How do I renew my lapsed operator qualification?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Emissions Guidelines and Compliance Times for Commercial and Industrial Solid Waste Incineration Units Model Rule-Operator Training...) For a lapse of less than 3 years, you must complete a standard annual refresher course described in...

  2. Are lapsed donors willing to resume blood donation, and what determines their motivation to do so?

    PubMed

    van Dongen, Anne; Abraham, Charles; Ruiter, Robert A C; Schaalma, Herman P; de Kort, Wim L A M; Dijkstra, J Anneke; Veldhuizen, Ingrid J T

    2012-06-01

    This study investigated the possibility of rerecruiting lapsed blood donors. Reasons for donation cessation, motivation to restart donation, and modifiable components of donation motivation were examined. We distinguished between lapsed donors who had passively withdrawn by merely not responding to donation invitations and donors who had contacted the blood bank to actively withdraw. A cross-sectional survey was sent to 400 actively lapsed donors and to 400 passively lapsed donors, measuring intention to restart donation and psychological correlates of restart intention. The data were analyzed using multiple regression analyses. The response rate among actively lapsed donors was higher than among passively lapsed donors (37% vs. 25%). Actively lapsed donors typically ceased donating because of physical reactions, while passively lapsed donors quit because of a busy lifestyle. Nonetheless, 51% of actively lapsed responders and 80% of passively lapsed responders were willing to restart donations. Multiple regression analysis showed that, for passively lapsed donors, cognitive attitude was the strongest correlate of intention to donate in the future (β=0.605, p<0.001), with affective attitude (β=0.239, p<0.05) and self-efficacy (β=0.266, p<0.001) explaining useful proportions of the variance as well. For actively lapsed donors, cognitive attitude was also the strongest correlate of intention (β=0.601, p<0.001), with affective attitude (β=0.345, p<0.001) and moral norm (β=-0.118, p<0.05) explaining smaller proportions of the variance. The majority of lapsed donors indicated a moderate to high intention to restart donations. Interventions focusing on boosting cognitive and affective attitudes and self-efficacy could further raise such intentions. © 2011 American Association of Blood Banks.

  3. Quantitative analysis of eosinophil chemotaxis tracked using a novel optical device -- TAXIScan.

    PubMed

    Nitta, Nao; Tsuchiya, Tomoko; Yamauchi, Akira; Tamatani, Takuya; Kanegasaki, Shiro

    2007-03-30

    We have reported previously the development of an optically accessible, horizontal chemotaxis apparatus, in which migration of cells in the channel from a start line can be traced with time-lapse intervals using a CCD camera (JIM 282, 1-11, 2003). To obtain statistical data of migrating cells, we have developed quantitative methods to calculate various parameters in the process of chemotaxis, employing human eosinophil and CXCL12 as a model cell and a model chemoattractant, respectively. Median values of velocity and directionality of each cell within an experimental period could be calculated from the migratory pathway data obtained from time-lapse images and the data were expressed as Velocity-Directionality (VD) plot. This plot is useful for quantitatively analyzing multiple migrating cells exposed to a certain chemoattractant, and can distinguish chemotaxis from random migration. Moreover precise observation of cell migration revealed that each cell had a different lag period before starting chemotaxis, indicating variation in cell sensitivity to the chemoattractant. Thus lag time of each cell before migration, and time course of increment of the migrating cell ratio at the early stages could be calculated. We also graphed decrement of still moving cell ratio at the later stages by calculating the duration time of cell migration of each cell. These graphs could distinguish different motion patterns of chemotaxis of eosinophils, in response to a range of chemoattractants; PGD(2), fMLP, CCL3, CCL5 and CXCL12. Finally, we compared parameters of eosinophils from normal volunteers, allergy patients and asthma patients and found significant difference in response to PGD(2). The quantitative methods described here could be applicable to image data obtained with any combination of cells and chemoattractants and useful not only for basic studies of chemotaxis but also for diagnosis and for drug screening.

  4. Individual and combined effects of multiple high-risk triggers on postcessation smoking urge and lapse.

    PubMed

    Lam, Cho Y; Businelle, Michael S; Aigner, Carrie J; McClure, Jennifer B; Cofta-Woerpel, Ludmila; Cinciripini, Paul M; Wetter, David W

    2014-05-01

    Negative affect, alcohol consumption, and presence of others smoking have consistently been implicated as risk factors for smoking lapse and relapse. What is not known, however, is how these factors work together to affect smoking outcomes. This paper uses ecological momentary assessment (EMA) collected during the first 7 days of a smoking cessation attempt to test the individual and combined effects of high-risk triggers on smoking urge and lapse. Participants were 300 female smokers who enrolled in a study that tested an individually tailored smoking cessation treatment. Participants completed EMA, which recorded negative affect, alcohol consumption, presence of others smoking, smoking urge, and smoking lapse, for 7 days starting on their quit date. Alcohol consumption, presence of others smoking, and negative affect were, independently and in combination, associated with increase in smoking urge and lapse. The results also found that the relationship between presence of others smoking and lapse and the relationship between negative affect and lapse were moderated by smoking urge. The current study found significant individual effects of alcohol consumption, presence of other smoking, and negative affect on smoking urge and lapse. Combing the triggers increased smoking urge and the risk for lapse to varying degrees, and the presence of all 3 triggers resulted in the highest urge and lapse risk.

  5. 77 FR 22069 - Proposed Information Collection (Notice of Lapse-Government Life Insurance); Comment Request

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-04-12

    ... of Lapse--Government Life Insurance); Comment Request AGENCY: Veterans Benefits Administration... determine claimants' eligibility to reinstate lapsed Government Life Insurance policy. DATES: Written.... Notice of Lapse--Government Life Insurance, VA Form 29-389. b. Application for Reinstatement, VA Form 29...

  6. Single-shot quantitative phase microscopy with color-multiplexed differential phase contrast (cDPC)

    PubMed Central

    2017-01-01

    We present a new technique for quantitative phase and amplitude microscopy from a single color image with coded illumination. Our system consists of a commercial brightfield microscope with one hardware modification—an inexpensive 3D printed condenser insert. The method, color-multiplexed Differential Phase Contrast (cDPC), is a single-shot variant of Differential Phase Contrast (DPC), which recovers the phase of a sample from images with asymmetric illumination. We employ partially coherent illumination to achieve resolution corresponding to 2× the objective NA. Quantitative phase can then be used to synthesize DIC and phase contrast images or extract shape and density. We demonstrate amplitude and phase recovery at camera-limited frame rates (50 fps) for various in vitro cell samples and c. elegans in a micro-fluidic channel. PMID:28152023

  7. Single-shot quantitative phase microscopy with color-multiplexed differential phase contrast (cDPC).

    PubMed

    Phillips, Zachary F; Chen, Michael; Waller, Laura

    2017-01-01

    We present a new technique for quantitative phase and amplitude microscopy from a single color image with coded illumination. Our system consists of a commercial brightfield microscope with one hardware modification-an inexpensive 3D printed condenser insert. The method, color-multiplexed Differential Phase Contrast (cDPC), is a single-shot variant of Differential Phase Contrast (DPC), which recovers the phase of a sample from images with asymmetric illumination. We employ partially coherent illumination to achieve resolution corresponding to 2× the objective NA. Quantitative phase can then be used to synthesize DIC and phase contrast images or extract shape and density. We demonstrate amplitude and phase recovery at camera-limited frame rates (50 fps) for various in vitro cell samples and c. elegans in a micro-fluidic channel.

  8. Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy.

    PubMed

    Doblas, Ana; Sánchez-Ortiga, Emilio; Martínez-Corral, Manuel; Saavedra, Genaro; Garcia-Sucerquia, Jorge

    2014-04-01

    The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more accurate QPI measurements with less number of computational operations.

  9. Quantitative measurements of nanoscale permittivity and conductivity using tuning-fork-based microwave impedance microscopy

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wu, Xiaoyu; Hao, Zhenqi; Wu, Di

    Here, we report quantitative measurements of nanoscale permittivity and conductivity using tuning-fork (TF) based microwave impedance microscopy (MIM). The system is operated under the driving amplitude modulation mode, which ensures satisfactory feedback stability on samples with rough surfaces. The demodulated MIM signals on a series of bulk dielectrics are in good agreement with results simulated by finite-element analysis. Using the TF-MIM, we have visualized the evolution of nanoscale conductance on back-gated MoS 2 field effect transistors, and the results are consistent with the transport data. Our work suggests that quantitative analysis of mesoscopic electrical properties can be achieved by near-fieldmore » microwave imaging with small distance modulation.« less

  10. Quantitative measurements of nanoscale permittivity and conductivity using tuning-fork-based microwave impedance microscopy

    DOE PAGES

    Wu, Xiaoyu; Hao, Zhenqi; Wu, Di; ...

    2018-04-01

    Here, we report quantitative measurements of nanoscale permittivity and conductivity using tuning-fork (TF) based microwave impedance microscopy (MIM). The system is operated under the driving amplitude modulation mode, which ensures satisfactory feedback stability on samples with rough surfaces. The demodulated MIM signals on a series of bulk dielectrics are in good agreement with results simulated by finite-element analysis. Using the TF-MIM, we have visualized the evolution of nanoscale conductance on back-gated MoS 2 field effect transistors, and the results are consistent with the transport data. Our work suggests that quantitative analysis of mesoscopic electrical properties can be achieved by near-fieldmore » microwave imaging with small distance modulation.« less

  11. Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography.

    PubMed

    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.

  12. Lab-On-Chip Clinorotation System for Live-Cell Microscopy Under Simulated Microgravity

    NASA Technical Reports Server (NTRS)

    Yew, Alvin G.; Atencia, Javier; Chinn, Ben; Hsieh, Adam H.

    2013-01-01

    Cells in microgravity are subject to mechanical unloading and changes to the surrounding chemical environment. How these factors jointly influence cellular function is not well understood. We can investigate their role using ground-based analogues to spaceflight, where mechanical unloading is simulated through the time-averaged nullification of gravity. The prevailing method for cellular microgravity simulation is to use fluid-filled containers called clinostats. However, conventional clinostats are not designed for temporally tracking cell response, nor are they able to establish dynamic fluid environments. To address these needs, we developed a Clinorotation Time-lapse Microscopy (CTM) system that accommodates lab-on- chip cell culture devices for visualizing time-dependent alterations to cellular behavior. For the purpose of demonstrating CTM, we present preliminary results showing time-dependent differences in cell area between human mesenchymal stem cells (hMSCs) under modeled microgravity and normal gravity.

  13. Lab-On-Chip Clinorotation System for Live-Cell Microscopy Under Simulated Microgravity

    NASA Technical Reports Server (NTRS)

    Yew, Alvin G.; Atencia, Javier; Chinn, Ben; Hsieh, Adam H.

    1980-01-01

    Cells in microgravity are subject to mechanical unloading and changes to the surrounding chemical environment. How these factors jointly influence cellular function is not well understood. We can investigate their role using ground-based analogues to spaceflight, where mechanical unloading is simulated through the time-averaged nullification of gravity. The prevailing method for cellular microgravity simulation is to use fluid-filled containers called clinostats. However, conventional clinostats are not designed for temporally tracking cell response, nor are they able to establish dynamic fluid environments. To address these needs, we developed a Clinorotation Time-lapse Microscopy (CTM) system that accommodates lab-on- chip cell culture devices for visualizing time-dependent alterations to cellular behavior. For the purpose of demonstrating CTM, we present preliminary results showing time-dependent differences in cell area between human mesenchymal stem cells (hMSCs) under modeled microgravity and normal gravity.

  14. Dual-view plane illumination microscopy for rapid and spatially isotropic imaging

    PubMed Central

    Kumar, Abhishek; Wu, Yicong; Christensen, Ryan; Chandris, Panagiotis; Gandler, William; McCreedy, Evan; Bokinsky, Alexandra; Colón-Ramos, Daniel A; Bao, Zhirong; McAuliffe, Matthew; Rondeau, Gary; Shroff, Hari

    2015-01-01

    We describe the construction and use of a compact dual-view inverted selective plane illumination microscope (diSPIM) for time-lapse volumetric (4D) imaging of living samples at subcellular resolution. Our protocol enables a biologist with some prior microscopy experience to assemble a diSPIM from commercially available parts, to align optics and test system performance, to prepare samples, and to control hardware and data processing with our software. Unlike existing light sheet microscopy protocols, our method does not require the sample to be embedded in agarose; instead, samples are prepared conventionally on glass coverslips. Tissue culture cells and Caenorhabditis elegans embryos are used as examples in this protocol; successful implementation of the protocol results in isotropic resolution and acquisition speeds up to several volumes per s on these samples. Assembling and verifying diSPIM performance takes ~6 d, sample preparation and data acquisition take up to 5 d and postprocessing takes 3–8 h, depending on the size of the data. PMID:25299154

  15. Sit still and pay attention: Using the Wii Balance-Board to detect lapses in concentration in children during psychophysical testing.

    PubMed

    Jones, Pete R

    2018-05-16

    During psychophysical testing, a loss of concentration can cause observers to answer incorrectly, even when the stimulus is clearly perceptible. Such lapses limit the accuracy and speed of many psychophysical measurements. This study evaluates an automated technique for detecting lapses based on body movement (postural instability). Thirty-five children (8-11 years of age) and 34 adults performed a typical psychophysical task (orientation discrimination) while seated on a Wii Fit Balance Board: a gaming device that measures center of pressure (CoP). Incorrect responses on suprathreshold catch trials provided the "reference standard" measure of when lapses in concentration occurred. Children exhibited significantly greater variability in CoP on lapse trials, indicating that postural instability provides a feasible, real-time index of concentration. Limitations and potential applications of this method are discussed.

  16. A time-lapse gravity survey of the Coso geothermal field, China Lake Naval Air Weapons Station, California

    USGS Publications Warehouse

    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.

  17. Ambient Seismic Noise Monitoring of Time-lapse Velocity Changes During CO2 Injection at Otway, South Australia

    NASA Astrophysics Data System (ADS)

    Saygin, E.; Lumley, D. E.

    2017-12-01

    We use continuous seismic data recorded with an array of 909 buried geophones at Otway, South Australia, to investigate the potential of using ambient seismic noise for time-lapse monitoring of the subsurface. The array was installed prior to a 15,000 ton CO2 injection in 2016-17, in order to detect and monitor the evolution of the injected CO2 plume, and any associated microseismic activity. Continuously recorded data from the vertical components of the geophone array were cross-correlated to retrieve the inter-station Green's functions. The dense collection of Green's functions contains diving body waves and surface Rayleigh waves. Green's Functions were then compared with each other at different time frames including the pre-injection period to track subtle changes in the travel times due to the CO2 injection. Our results show a clear change in the velocities of Green's functions at the start of injection for both body waves and surface waves for wave paths traversing the injection area, whereas the observed changes are much smaller for areas which are far from the injection well.

  18. Real-time high dynamic range laser scanning microscopy

    PubMed Central

    Vinegoni, C.; Leon Swisher, C.; Fumene Feruglio, P.; Giedt, R. J.; Rousso, D. L.; Stapleton, S.; Weissleder, R.

    2016-01-01

    In conventional confocal/multiphoton fluorescence microscopy, images are typically acquired under ideal settings and after extensive optimization of parameters for a given structure or feature, often resulting in information loss from other image attributes. To overcome the problem of selective data display, we developed a new method that extends the imaging dynamic range in optical microscopy and improves the signal-to-noise ratio. Here we demonstrate how real-time and sequential high dynamic range microscopy facilitates automated three-dimensional neural segmentation. We address reconstruction and segmentation performance on samples with different size, anatomy and complexity. Finally, in vivo real-time high dynamic range imaging is also demonstrated, making the technique particularly relevant for longitudinal imaging in the presence of physiological motion and/or for quantification of in vivo fast tracer kinetics during functional imaging. PMID:27032979

  19. Analysis of Snow Line and Albedo Conditions By Means of Time-Lapse Photography on Tapado Glacier, Chile

    NASA Astrophysics Data System (ADS)

    Vivero, S.; MacDonell, S.; McPhee, J. P.

    2014-12-01

    In the semiarid Coquimbo Region of Chile, high-altitude glaciers and seasonal snow are important sources of freshwater for irrigated agriculture and urban consumption. Due to the aridity of the environment, losses due to sublimation are large which means that accurate melt modelling is essential in order to reliably estimate streamflow. Since 2008, the CEAZA glaciology group has been studying the energy and mass balance of the largest glacier in the catchment, the Tapado Glacier using field and remote sensing measurements, and numerical modelling. The Tapado glacier system (30°08' S, 69°55' W) is a complex assemblage of uncovered and debris-covered ice located at the head of the Elqui basin between 4500 and 5536 m a.s.l. Energy balance modelling studies at the site have been limited in scope due to the development of ice pinnacles or penitentes on snow and ice surfaces. These features complicate energy distribution across the surface, due to modifications of parameters such as albedo. In this paper, we use time-lapse photography and automatic weather station (AWS) measurements to investigate how the development of penitentes impacts the spatial and temporal variability of albedo across the glacier surface and whether terrestrial photography is appropriate for use at such locations. Oblique photographs obtained from a high vantage point were georeferenced using a high resolution digital elevation model available for the entire glacier and its environs. By comparing the photographic data with point albedo measurements made at an AWS, distributed albedo maps were produced. Preliminary results suggest that distributed albedo values may be underestimated by the formation and development of penitentes during the ablation season. Moreover, it was observed that the evolution of the snow line during summer was not only topographically controlled but also modified by occasional convective snowfalls. Time-lapse photography provided to be a cost-effective tool for monitoring

  20. Time-lapse ERT and DTS for seasonal and short-term monitoring of an alpine river hyporheic zone

    NASA Astrophysics Data System (ADS)

    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

  1. Quantitative pathology in virtual microscopy: history, applications, perspectives.

    PubMed

    Kayser, Gian; Kayser, Klaus

    2013-07-01

    With the emerging success of commercially available personal computers and the rapid progress in the development of information technologies, morphometric analyses of static histological images have been introduced to improve our understanding of the biology of diseases such as cancer. First applications have been quantifications of immunohistochemical expression patterns. In addition to object counting and feature extraction, laws of thermodynamics have been applied in morphometric calculations termed syntactic structure analysis. Here, one has to consider that the information of an image can be calculated for separate hierarchical layers such as single pixels, cluster of pixels, segmented small objects, clusters of small objects, objects of higher order composed of several small objects. Using syntactic structure analysis in histological images, functional states can be extracted and efficiency of labor in tissues can be quantified. Image standardization procedures, such as shading correction and color normalization, can overcome artifacts blurring clear thresholds. Morphometric techniques are not only useful to learn more about biological features of growth patterns, they can also be helpful in routine diagnostic pathology. In such cases, entropy calculations are applied in analogy to theoretical considerations concerning information content. Thus, regions with high information content can automatically be highlighted. Analysis of the "regions of high diagnostic value" can deliver in the context of clinical information, site of involvement and patient data (e.g. age, sex), support in histopathological differential diagnoses. It can be expected that quantitative virtual microscopy will open new possibilities for automated histological support. Automated integrated quantification of histological slides also serves for quality assurance. The development and theoretical background of morphometric analyses in histopathology are reviewed, as well as their application

  2. Time-stretch microscopy based on time-wavelength sequence reconstruction from wideband incoherent source

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zhang, Chi, E-mail: chizheung@gmail.com; Xu, Yiqing; Wei, Xiaoming

    2014-07-28

    Time-stretch microscopy has emerged as an ultrafast optical imaging concept offering the unprecedented combination of the imaging speed and sensitivity. However, dedicated wideband and coherence optical pulse source with high shot-to-shot stability has been mandated for time-wavelength mapping—the enabling process for ultrahigh speed wavelength-encoded image retrieval. From the practical point of view, exploiting methods to relax the stringent requirements (e.g., temporal stability and coherence) for the source of time-stretch microscopy is thus of great value. In this paper, we demonstrated time-stretch microscopy by reconstructing the time-wavelength mapping sequence from a wideband incoherent source. Utilizing the time-lens focusing mechanism mediated bymore » a narrow-band pulse source, this approach allows generation of a wideband incoherent source, with the spectral efficiency enhanced by a factor of 18. As a proof-of-principle demonstration, time-stretch imaging with the scan rate as high as MHz and diffraction-limited resolution is achieved based on the wideband incoherent source. We note that the concept of time-wavelength sequence reconstruction from wideband incoherent source can also be generalized to any high-speed optical real-time measurements, where wavelength is acted as the information carrier.« less

  3. Combined use of X-ray fluorescence microscopy, phase contrast imaging for high resolution quantitative iron mapping in inflamed cells

    NASA Astrophysics Data System (ADS)

    Gramaccioni, C.; Procopio, A.; Farruggia, G.; Malucelli, E.; Iotti, S.; Notargiacomo, A.; Fratini, M.; Yang, Y.; Pacureanu, A.; Cloetens, P.; Bohic, S.; Massimi, L.; Cutone, A.; Valenti, P.; Rosa, L.; Berlutti, F.; Lagomarsino, S.

    2017-06-01

    X-ray fluorescence microscopy (XRFM) is a powerful technique to detect and localize elements in cells. To derive information useful for biology and medicine, it is essential not only to localize, but also to map quantitatively the element concentration. Here we applied quantitative XRFM to iron in phagocytic cells. Iron, a primary component of living cells, can become toxic when present in excess. In human fluids, free iron is maintained at 10-18 M concentration thanks to iron binding proteins as lactoferrin (Lf). The iron homeostasis, involving the physiological ratio of iron between tissues/secretions and blood, is strictly regulated by ferroportin, the sole protein able to export iron from cells to blood. Inflammatory processes induced by lipopolysaccharide (LPS) or bacterial pathoge inhibit ferroportin synthesis in epithelial and phagocytic cells thus hindering iron export, increasing intracellular iron and bacterial multiplication. In this respect, Lf is emerging as an important regulator of both iron and inflammatory homeostasis. Here we studied phagocytic cells inflamed by bacterial LPS and untreated or treated with milk derived bovine Lf. Quantitative mapping of iron concentration and mass fraction at high spatial resolution is obtained combining X-ray fluorescence microscopy, atomic force microscopy and synchrotron phase contrast imaging.

  4. New tools for comparing microscopy images: quantitative analysis of cell types in Bacillus subtilis.

    PubMed

    van Gestel, Jordi; Vlamakis, Hera; Kolter, Roberto

    2015-02-15

    Fluorescence microscopy is a method commonly used to examine individual differences between bacterial cells, yet many studies still lack a quantitative analysis of fluorescence microscopy data. Here we introduce some simple tools that microbiologists can use to analyze and compare their microscopy images. We show how image data can be converted to distribution data. These data can be subjected to a cluster analysis that makes it possible to objectively compare microscopy images. The distribution data can further be analyzed using distribution fitting. We illustrate our methods by scrutinizing two independently acquired data sets, each containing microscopy images of a doubly labeled Bacillus subtilis strain. For the first data set, we examined the expression of srfA and tapA, two genes which are expressed in surfactin-producing and matrix-producing cells, respectively. For the second data set, we examined the expression of eps and tapA; these genes are expressed in matrix-producing cells. We show that srfA is expressed by all cells in the population, a finding which contrasts with a previously reported bimodal distribution of srfA expression. In addition, we show that eps and tapA do not always have the same expression profiles, despite being expressed in the same cell type: both operons are expressed in cell chains, while single cells mainly express eps. These findings exemplify that the quantification and comparison of microscopy data can yield insights that otherwise would go unnoticed. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  5. In-focal-plane characterization of excitation distribution for quantitative fluorescence microscopy applications

    NASA Astrophysics Data System (ADS)

    Dietrich, Klaus; Brülisauer, Martina; ćaǧin, Emine; Bertsch, Dietmar; Lüthi, Stefan; Heeb, Peter; Stärker, Ulrich; Bernard, André

    2017-06-01

    The applications of fluorescence microscopy span medical diagnostics, bioengineering and biomaterial analytics. Full exploitation of fluorescent microscopy is hampered by imperfections in illumination, detection and filtering. Mainly, errors stem from deviations induced by real-world components inducing spatial or angular variations of propagation properties along the optical path, and they can be addressed through consistent and accurate calibration. For many applications, uniform signal to noise ratio (SNR) over the imaging area is required. Homogeneous SNR can be achieved by quantifying and compensating for the signal bias. We present a method to quantitatively characterize novel reference materials as a calibration reference for biomaterials analytics. The reference materials under investigation comprise thin layers of fluorophores embedded in polymer matrices. These layers are highly homogeneous in their fluorescence response, where cumulative variations do not exceed 1% over the field of view (1.5 x 1.1 mm). An automated and reproducible measurement methodology, enabling sufficient correction for measurement artefacts, is reported. The measurement setup is equipped with an autofocus system, ensuring that the measured film quality is not artificially increased by out-of-focus reduction of the system modulation transfer function. The quantitative characterization method is suitable for analysis of modified bio-materials, especially through patterned protein decoration. The imaging method presented here can be used to statistically analyze protein patterns, thereby increasing both precision and throughput. Further, the method can be developed to include a reference emitter and detector pair on the image surface of the reference object, in order to provide traceable measurements.

  6. Transformation and motility of human platelets: details of the shape change and release reaction observed by optical and electron microscopy

    PubMed Central

    1979-01-01

    Blood platelets from 10 normal human subjects have been examined with a sensitive differential interference contrast (DIC) microscope. The entire transformation process during adhesion to glass is clearly visible and has been recorded cinematographically, including the disk to sphere change of shape, the formation of sessile protuberances, the extension and retraction of pseudopodia, and the spreading, ruffling, and occasional regression of the hyalomere. The exocytosis of intact dense bodies can be observed either by DIC microscopy, or by epifluorescence microscopy in platelets stained with mepacrine. Details of fluorescent flashes indicate that the dense bodies usually release their contents extracellularly, may do so intracytoplasmically under the influence of strong, short wavelength light on some preparations of mepacrine-stained platelets. The release of one or more dense bodies leaves a crater of variable size on the upper surface of the granulomere. Such craters represent the surface component of the open canalicular system and their formation and disappearance can be directly observed. Because these techniques permit quantitation of several parameters of motility which are not readily observable by other techniques, it is suggested that high extinction DIC microscope examination may become a rapid and useful method of studying congenital and acquired platelet disorders. Many features of platelet transformation have been confirmed and extended by scanning electron micrographs. These can in turn be interpreted by reference to time- lapse films of living platelets. PMID:511936

  7. Predictive modeling of addiction lapses in a mobile health application.

    PubMed

    Chih, Ming-Yuan; Patton, Timothy; McTavish, Fiona M; Isham, Andrew J; Judkins-Fisher, Chris L; Atwood, Amy K; Gustafson, David H

    2014-01-01

    The chronically relapsing nature of alcoholism leads to substantial personal, family, and societal costs. Addiction-comprehensive health enhancement support system (A-CHESS) is a smartphone application that aims to reduce relapse. To offer targeted support to patients who are at risk of lapses within the coming week, a Bayesian network model to predict such events was constructed using responses on 2,934 weekly surveys (called the Weekly Check-in) from 152 alcohol-dependent individuals who recently completed residential treatment. The Weekly Check-in is a self-monitoring service, provided in A-CHESS, to track patients' recovery progress. The model showed good predictability, with the area under receiver operating characteristic curve of 0.829 in the 10-fold cross-validation and 0.912 in the external validation. The sensitivity/specificity table assists the tradeoff decisions necessary to apply the model in practice. This study moves us closer to the goal of providing lapse prediction so that patients might receive more targeted and timely support. © 2013.

  8. Predictive Modeling of Addiction Lapses in a Mobile Health Application

    PubMed Central

    Chih, Ming-Yuan; Patton, Timothy; McTavish, Fiona M.; Isham, Andrew; Judkins-Fisher, Chris L.; Atwood, Amy K.; Gustafson, David H.

    2013-01-01

    The chronically relapsing nature of alcoholism leads to substantial personal, family, and societal costs. Addiction-Comprehensive Health Enhancement Support System (A-CHESS) is a smartphone application that aims to reduce relapse. To offer targeted support to patients who are at risk of lapses within the coming week, a Bayesian network model to predict such events was constructed using responses on 2,934 weekly surveys (called the Weekly Check-in) from 152 alcohol-dependent individuals who recently completed residential treatment. The Weekly Check-in is a self-monitoring service, provided in A-CHESS, to track patients’ recovery progress. The model showed good predictability, with the area under receiver operating characteristic curve of 0.829 in the 10-fold cross-validation and 0.912 in the external validation. The sensitivity/specificity table assists the tradeoff decisions necessary to apply the model in practice. This study moves us closer to the goal of providing lapse prediction so that patients might receive more targeted and timely support. PMID:24035143

  9. Analysis of interphase node proteins in fission yeast by quantitative and superresolution fluorescence microscopy

    PubMed Central

    Akamatsu, Matthew; Lin, Yu; Bewersdorf, Joerg; Pollard, Thomas D.

    2017-01-01

    We used quantitative confocal microscopy and FPALM superresolution microscopy of live fission yeast to investigate the structures and assembly of two types of interphase nodes—multiprotein complexes associated with the plasma membrane that merge together and mature into the precursors of the cytokinetic contractile ring. During the long G2 phase of the cell cycle, seven different interphase node proteins maintain constant concentrations as they accumulate in proportion to cell volume. During mitosis, the total numbers of type 1 node proteins (cell cycle kinases Cdr1p, Cdr2p, Wee1p, and anillin Mid1p) are constant even when the nodes disassemble. Quantitative measurements provide strong evidence that both types of nodes have defined sizes and numbers of constituent proteins, as observed for cytokinesis nodes. Type 1 nodes assemble in two phases—a burst at the end of mitosis, followed by steady increase during interphase to double the initial number. Type 2 nodes containing Blt1p, Rho-GEF Gef2p, and kinesin Klp8p remain intact throughout the cell cycle and are constituents of the contractile ring. They are released from the contractile ring as it disassembles and then associate with type 1 nodes around the equator of the cell during interphase. PMID:28539404

  10. Quantitative Analysis of Subcellular Distribution of the SUMO Conjugation System by Confocal Microscopy Imaging.

    PubMed

    Mas, Abraham; Amenós, Montse; Lois, L Maria

    2016-01-01

    Different studies point to an enrichment in SUMO conjugation in the cell nucleus, although non-nuclear SUMO targets also exist. In general, the study of subcellular localization of proteins is essential for understanding their function within a cell. Fluorescence microscopy is a powerful tool for studying subcellular protein partitioning in living cells, since fluorescent proteins can be fused to proteins of interest to determine their localization. Subcellular distribution of proteins can be influenced by binding to other biomolecules and by posttranslational modifications. Sometimes these changes affect only a portion of the protein pool or have a partial effect, and a quantitative evaluation of fluorescence images is required to identify protein redistribution among subcellular compartments. In order to obtain accurate data about the relative subcellular distribution of SUMO conjugation machinery members, and to identify the molecular determinants involved in their localization, we have applied quantitative confocal microscopy imaging. In this chapter, we will describe the fluorescent protein fusions used in these experiments, and how to measure, evaluate, and compare average fluorescence intensities in cellular compartments by image-based analysis. We show the distribution of some components of the Arabidopsis SUMOylation machinery in epidermal onion cells and how they change their distribution in the presence of interacting partners or even when its activity is affected.

  11. Bessel light sheet structured illumination microscopy

    NASA Astrophysics Data System (ADS)

    Noshirvani Allahabadi, Golchehr

    Biomedical study researchers using animals to model disease and treatment need fast, deep, noninvasive, and inexpensive multi-channel imaging methods. Traditional fluorescence microscopy meets those criteria to an extent. Specifically, two-photon and confocal microscopy, the two most commonly used methods, are limited in penetration depth, cost, resolution, and field of view. In addition, two-photon microscopy has limited ability in multi-channel imaging. Light sheet microscopy, a fast developing 3D fluorescence imaging method, offers attractive advantages over traditional two-photon and confocal microscopy. Light sheet microscopy is much more applicable for in vivo 3D time-lapsed imaging, owing to its selective illumination of tissue layer, superior speed, low light exposure, high penetration depth, and low levels of photobleaching. However, standard light sheet microscopy using Gaussian beam excitation has two main disadvantages: 1) the field of view (FOV) of light sheet microscopy is limited by the depth of focus of the Gaussian beam. 2) Light-sheet images can be degraded by scattering, which limits the penetration of the excitation beam and blurs emission images in deep tissue layers. While two-sided sheet illumination, which doubles the field of view by illuminating the sample from opposite sides, offers a potential solution, the technique adds complexity and cost to the imaging system. We investigate a new technique to address these limitations: Bessel light sheet microscopy in combination with incoherent nonlinear Structured Illumination Microscopy (SIM). Results demonstrate that, at visible wavelengths, Bessel excitation penetrates up to 250 microns deep in the scattering media with single-side illumination. Bessel light sheet microscope achieves confocal level resolution at a lateral resolution of 0.3 micron and an axial resolution of 1 micron. Incoherent nonlinear SIM further reduces the diffused background in Bessel light sheet images, resulting in

  12. Fast time-resolved electrostatic force microscopy: Achieving sub-cycle time resolution

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Karatay, Durmus U.; Harrison, Jeffrey S.; Glaz, Micah S.

    The ability to measure microsecond- and nanosecond-scale local dynamics below the diffraction limit with widely available atomic force microscopy hardware would enable new scientific studies in fields ranging from biology to semiconductor physics. However, commercially available scanning-probe instruments typically offer the ability to measure dynamics only on time scales of milliseconds to seconds. Here, we describe in detail the implementation of fast time-resolved electrostatic force microscopy using an oscillating cantilever as a means to measure fast local dynamics following a perturbation to a sample. We show how the phase of the oscillating cantilever relative to the perturbation event is criticalmore » to achieving reliable sub-cycle time resolution. We explore how noise affects the achievable time resolution and present empirical guidelines for reducing noise and optimizing experimental parameters. Specifically, we show that reducing the noise on the cantilever by using photothermal excitation instead of piezoacoustic excitation further improves time resolution. We demonstrate the discrimination of signal rise times with time constants as fast as 10 ns, and simultaneous data acquisition and analysis for dramatically improved image acquisition times.« less

  13. Development of fluorescent glucose bioprobes and their application on real-time and quantitative monitoring of glucose uptake in living cells.

    PubMed

    Lee, Hyang Yeon; Lee, Jae Jeong; Park, Jongmin; Park, Seung Bum

    2011-01-03

    We developed a novel fluorescent glucose bioprobe, GB2-Cy3, for the real-time and quantitative monitoring of glucose uptake in living cells. We synthesized a series of fluorescent glucose analogues by adding Cy3 fluorophores to the α-anomeric position of D-glucose through various linkers. Systematic and quantitative analysis of these Cy3-labeled glucose analogues revealed that GB2-Cy3 was the ideal fluorescent glucose bioprobe. The cellular uptake of this probe competed with the cellular uptake of D-glucose in the media and was mediated by a glucose-specific transport system, and not by passive diffusion. Flow cytometry and fluorescence microscopy analyses revealed that GB2-Cy3 is ten times more sensitive than 2-NBDG, a leading fluorescent glucose bioprobe. GB2-Cy3 can also be utilized for the quantitative flow cytometry monitoring of glucose uptake in metabolically active C2C12 myocytes under various treatment conditions. As opposed to a glucose uptake assay performed by using radioisotope-labeled deoxy-D-glucose and a scintillation counter, GB2-Cy3 allows the real-time monitoring of glucose uptake in living cells under various experimental conditions by using fluorescence microscopy or confocal laser scanning microscopy (CLSM). Therefore, we believe that GB2-Cy3 can be utilized in high-content screening (HCS) for the discovery of novel therapeutic agents and for making significant advances in biomedical studies and diagnosis of various diseases, especially metabolic diseases. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Tracing cell lineages in videos of lens-free microscopy.

    PubMed

    Rempfler, Markus; Stierle, Valentin; Ditzel, Konstantin; Kumar, Sanjeev; Paulitschke, Philipp; Andres, Bjoern; Menze, Bjoern H

    2018-06-05

    In vitro experiments with cultured cells are essential for studying their growth and migration pattern and thus, for gaining a better understanding of cancer progression and its treatment. Recent progress in lens-free microscopy (LFM) has rendered it an inexpensive tool for label-free, continuous live cell imaging, yet there is only little work on analysing such time-lapse image sequences. We propose (1) a cell detector for LFM images based on fully convolutional networks and residual learning, and (2) a probabilistic model based on moral lineage tracing that explicitly handles multiple detections and temporal successor hypotheses by clustering and tracking simultaneously. (3) We benchmark our method in terms of detection and tracking scores on a dataset of three annotated sequences of several hours of LFM, where we demonstrate our method to produce high quality lineages. (4) We evaluate its performance on a somewhat more challenging problem: estimating cell lineages from the LFM sequence as would be possible from a corresponding fluorescence microscopy sequence. We present experiments on 16 LFM sequences for which we acquired fluorescence microscopy in parallel and generated annotations from them. Finally, (5) we showcase our methods effectiveness for quantifying cell dynamics in an experiment with skin cancer cells. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Using digital inline holographic microscopy and quantitative phase contrast imaging to assess viability of cultured mammalian cells

    NASA Astrophysics Data System (ADS)

    Missan, Sergey; Hrytsenko, Olga

    2015-03-01

    Digital inline holographic microscopy was used to record holograms of mammalian cells (HEK293, B16, and E0771) in culture. The holograms have been reconstructed using Octopus software (4Deep inwater imaging) and phase shift maps were unwrapped using the FFT-based phase unwrapping algorithm. The unwrapped phase shifts were used to determine the maximum phase shifts in individual cells. Addition of 0.5 mM H2O2 to cell media produced rapid rounding of cultured cells, followed by cell membrane rupture. The cell morphology changes and cell membrane ruptures were detected in real time and were apparent in the unwrapped phase shift images. The results indicate that quantitative phase contrast imaging produced by the digital inline holographic microscope can be used for the label-free real time automated determination of cell viability and confluence in mammalian cell cultures.

  16. Dynamic Association between Negative Affect and Alcohol Lapses following Alcohol Treatment

    ERIC Educational Resources Information Center

    Witkiewitz, Katie; Villarroel, Nadia Aracelliz

    2009-01-01

    Clinical research has found a strong association between negative affect and returning to alcohol use after a period of abstinence. Yet little is known about the probability of a lapse given a particular level of negative affect or whether there is a reciprocal relationship between negative affect and alcohol use across time. The goal of the…

  17. Mathematical imaging methods for mitosis analysis in live-cell phase contrast microscopy.

    PubMed

    Grah, Joana Sarah; Harrington, Jennifer Alison; Koh, Siang Boon; Pike, Jeremy Andrew; Schreiner, Alexander; Burger, Martin; Schönlieb, Carola-Bibiane; Reichelt, Stefanie

    2017-02-15

    In this paper we propose a workflow to detect and track mitotic cells in time-lapse microscopy image sequences. In order to avoid the requirement for cell lines expressing fluorescent markers and the associated phototoxicity, phase contrast microscopy is often preferred over fluorescence microscopy in live-cell imaging. However, common specific image characteristics complicate image processing and impede use of standard methods. Nevertheless, automated analysis is desirable due to manual analysis being subjective, biased and extremely time-consuming for large data sets. Here, we present the following workflow based on mathematical imaging methods. In the first step, mitosis detection is performed by means of the circular Hough transform. The obtained circular contour subsequently serves as an initialisation for the tracking algorithm based on variational methods. It is sub-divided into two parts: in order to determine the beginning of the whole mitosis cycle, a backwards tracking procedure is performed. After that, the cell is tracked forwards in time until the end of mitosis. As a result, the average of mitosis duration and ratios of different cell fates (cell death, no division, division into two or more daughter cells) can be measured and statistics on cell morphologies can be obtained. All of the tools are featured in the user-friendly MATLAB®Graphical User Interface MitosisAnalyser. Copyright © 2017. Published by Elsevier Inc.

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

    NASA Astrophysics Data System (ADS)

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

    2012-03-01

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

  19. Long-term High-Resolution Intravital Microscopy in the Lung with a Vacuum Stabilized Imaging Window

    PubMed Central

    Rodriguez-Tirado, Carolina; Kitamura, Takanori; Kato, Yu; Pollard, Jeffery W.; Condeelis, John S.; Entenberg, David

    2017-01-01

    Metastasis to secondary sites such as the lung, liver and bone is a traumatic event with a mortality rate of approximately 90% 1. Of these sites, the lung is the most difficult to assess using intravital optical imaging due to its enclosed position within the body, delicate nature and vital role in sustaining proper physiology. While clinical modalities (positron emission tomography (PET), magnetic resonance imaging (MRI) and computed tomography (CT)) are capable of providing noninvasive images of this tissue, they lack the resolution necessary to visualize the earliest seeding events, with a single pixel consisting of nearly a thousand cells. Current models of metastatic lung seeding postulate that events just after a tumor cell's arrival are deterministic for survival and subsequent growth. This means that real-time intravital imaging tools with single cell resolution 2 are required in order to define the phenotypes of the seeding cells and test these models. While high resolution optical imaging of the lung has been performed using various ex vivo preparations, these experiments are typically single time-point assays and are susceptible to artifacts and possible erroneous conclusions due to the dramatically altered environment (temperature, profusion, cytokines, etc.) resulting from removal from the chest cavity and circulatory system 3. Recent work has shown that time-lapse intravital optical imaging of the intact lung is possible using a vacuum stabilized imaging window 2,4,5 however, typical imaging times have been limited to approximately 6 hr. Here we describe a protocol for performing long-term intravital time-lapse imaging of the lung utilizing such a window over a period of 12 hr. The time-lapse image sequences obtained using this method enable visualization and quantitation of cell-cell interactions, membrane dynamics and vascular perfusion in the lung. We further describe an image processing technique that gives an unprecedentedly clear view of the

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

    PubMed

    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.