771 research outputs found

    Integrating remote sensing datasets into ecological modelling: a Bayesian approach

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    Process-based models have been used to simulate 3-dimensional complexities of forest ecosystems and their temporal changes, but their extensive data requirement and complex parameterisation have often limited their use for practical management applications. Increasingly, information retrieved using remote sensing techniques can help in model parameterisation and data collection by providing spatially and temporally resolved forest information. In this paper, we illustrate the potential of Bayesian calibration for integrating such data sources to simulate forest production. As an example, we use the 3-PG model combined with hyperspectral, LiDAR, SAR and field-based data to simulate the growth of UK Corsican pine stands. Hyperspectral, LiDAR and SAR data are used to estimate LAI dynamics, tree height and above ground biomass, respectively, while the Bayesian calibration provides estimates of uncertainties to model parameters and outputs. The Bayesian calibration contrasts with goodness-of-fit approaches, which do not provide uncertainties to parameters and model outputs. Parameters and the data used in the calibration process are presented in the form of probability distributions, reflecting our degree of certainty about them. After the calibration, the distributions are updated. To approximate posterior distributions (of outputs and parameters), a Markov Chain Monte Carlo sampling approach is used (25 000 steps). A sensitivity analysis is also conducted between parameters and outputs. Overall, the results illustrate the potential of a Bayesian framework for truly integrative work, both in the consideration of field-based and remotely sensed datasets available and in estimating parameter and model output uncertainties

    Visualizing Single-molecule DNA Replication with Fluorescence Microscopy

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    We describe a simple fluorescence microscopy-based real-time method for observing DNA replication at the single-molecule level. A circular, forked DNA template is attached to a functionalized glass coverslip and replicated extensively after introduction of replication proteins and nucleotides (Figure 1). The growing product double-strand DNA (dsDNA) is extended with laminar flow and visualized by using an intercalating dye. Measuring the position of the growing DNA end in real time allows precise determination of replication rate (Figure 2). Furthermore, the length of completed DNA products reports on the processivity of replication. This experiment can be performed very easily and rapidly and requires only a fluorescence microscope with a reasonably sensitive camera

    Droplet microfluidics: a tool for biology, chemistry and nanotechnology

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    The ability to perform laboratory operations on small scales using miniaturized devices provides numerous benefits, including reduced quantities of reagents and waste as well as increased portability and controllability of assays. These operations can involve reaction components in the solution phase and as a result, their miniaturization can be accomplished through microfluidic approaches. One such approach, droplet microfluidics, provides a high-throughput platform for a wide range of assays and approaches in chemistry, biology and nanotechnology. We highlight recent advances in the application of droplet microfluidics in chip-based technologies, such as single-cell analysis tools, small-scale cell cultures, in-droplet chemical synthesis, high-throughput drug screening, and nanodevice fabrication

    Inferring DNA sequences from mechanical unzipping data: the large-bandwidth case

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    The complementary strands of DNA molecules can be separated when stretched apart by a force; the unzipping signal is correlated to the base content of the sequence but is affected by thermal and instrumental noise. We consider here the ideal case where opening events are known to a very good time resolution (very large bandwidth), and study how the sequence can be reconstructed from the unzipping data. Our approach relies on the use of statistical Bayesian inference and of Viterbi decoding algorithm. Performances are studied numerically on Monte Carlo generated data, and analytically. We show how multiple unzippings of the same molecule may be exploited to improve the quality of the prediction, and calculate analytically the number of required unzippings as a function of the bandwidth, the sequence content, the elasticity parameters of the unzipped strands

    Direct Observation of Enzymes Replicating DNA Using a Single-molecule DNA Stretching Assay

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    We describe a method for observing real time replication of individual DNA molecules mediated by proteins of the bacteriophage replication system. Linearized λ DNA is modified to have a biotin on the end of one strand, and a digoxigenin moiety on the other end of the same strand. The biotinylated end is attached to a functionalized glass coverslip and the digoxigeninated end to a small bead. The assembly of these DNA-bead tethers on the surface of a flow cell allows a laminar flow to be applied to exert a drag force on the bead. As a result, the DNA is stretched close to and parallel to the surface of the coverslip at a force that is determined by the flow rate (Figure 1). The length of the DNA is measured by monitoring the position of the bead. Length differences between single- and double-stranded DNA are utilized to obtain real-time information on the activity of the replication proteins at the fork. Measuring the position of the bead allows precise determination of the rates and processivities of DNA unwinding and polymerization (Figure 2)
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