28 resultados para Subcellular localisation


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Many problems in control and signal processing can be formulated as sequential decision problems for general state space models. However, except for some simple models one cannot obtain analytical solutions and has to resort to approximation. In this thesis, we have investigated problems where Sequential Monte Carlo (SMC) methods can be combined with a gradient based search to provide solutions to online optimisation problems. We summarise the main contributions of the thesis as follows. Chapter 4 focuses on solving the sensor scheduling problem when cast as a controlled Hidden Markov Model. We consider the case in which the state, observation and action spaces are continuous. This general case is important as it is the natural framework for many applications. In sensor scheduling, our aim is to minimise the variance of the estimation error of the hidden state with respect to the action sequence. We present a novel SMC method that uses a stochastic gradient algorithm to find optimal actions. This is in contrast to existing works in the literature that only solve approximations to the original problem. In Chapter 5 we presented how an SMC can be used to solve a risk sensitive control problem. We adopt the use of the Feynman-Kac representation of a controlled Markov chain flow and exploit the properties of the logarithmic Lyapunov exponent, which lead to a policy gradient solution for the parameterised problem. The resulting SMC algorithm follows a similar structure with the Recursive Maximum Likelihood(RML) algorithm for online parameter estimation. In Chapters 6, 7 and 8, dynamic Graphical models were combined with with state space models for the purpose of online decentralised inference. We have concentrated more on the distributed parameter estimation problem using two Maximum Likelihood techniques, namely Recursive Maximum Likelihood (RML) and Expectation Maximization (EM). The resulting algorithms can be interpreted as an extension of the Belief Propagation (BP) algorithm to compute likelihood gradients. In order to design an SMC algorithm, in Chapter 8 uses a nonparametric approximations for Belief Propagation. The algorithms were successfully applied to solve the sensor localisation problem for sensor networks of small and medium size.

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The impact response of laminated composites consisting of alternate layers of AI ahoy foam and Al2O3 was studied experimentally in low and intermediate velocity regimes. Low velocity impacts (1.2-2.8 m s(-1)) were conducted using an instrumented falling weight apparatus and were compared with static indentation tests (0.2 x 10(-4) m s(-1)). Intermediate velocity impacts were carried out by means of both Hopkinson bar (60 m s(-1)) and gas gun (200 m s(-1)) tests, Post-impact damage was assessed using X-ray radiography and microscopy, It was found that there is good correlation between low velocity impact and quasi-static responses. In both cases, penetration of the layered targets resulted in the formation of a distinctive plug. Increasing impact velocity (intermediate velocity range) snitched the penetration mode from plugging to fragmentation, giving rise to an increase in the absorbed energy. In this range, impacts led to localisation of damage in the region under the projectile, Furthermore, a comparison has been made between the penetration response of foam laminates and dense metal laminates of equivalent areal density. Preliminary results suggest that the dense metal laminates are superseded by the foam laminates on an energy absorption basis.

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We introduce a new algorithm to automatically identify the time and pixel location of foot contact events in high speed video of sprinters. We use this information to autonomously synchronise and overlay multiple recorded performances to provide feedback to athletes and coaches during their training sessions. The algorithm exploits the variation in speed of different parts of the body during sprinting. We use an array of foreground accumulators to identify short-term static pixels and a temporal analysis of the associated static regions to identify foot contacts. We evaluated the technique using 13 videos of three sprinters. It successfully identifed 55 of the 56 contacts, with a mean localisation error of 1.39±1.05 pixels. Some videos were also seen to produce additional, spurious contacts. We present heuristics to help identify the true contacts. © 2011 Springer-Verlag Berlin Heidelberg.

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Super-Resolution imaging techniques such as Fluorescent Photo-Activation Localisation Microscopy (FPALM) have created a powerful new toolkit for investigating living cells, however a simple platform for growing, trapping, holding and controlling the cells is needed before the approach can become truly widespread. We present a microfluidic device formed in polydimethylsiloxane (PDMS) with a fluidic design which traps cells in a high-density array of wells and holds them very still throughout the life cycle, using hydrodynamic forces only. The device meets or exceeds all the necessary criteria for FPALM imaging of Schizosaccharomyces pombe and is designed to remain flexible, robust and easy to use. © 2011 IEEE.

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One-cell-thick monolayers are the simplest tissues in multicellular organisms, yet they fulfill critical roles in development and normal physiology. In early development, embryonic morphogenesis results largely from monolayer rearrangement and deformation due to internally generated forces. Later, monolayers act as physical barriers separating the internal environment from the exterior and must withstand externally applied forces. Though resisting and generating mechanical forces is an essential part of monolayer function, simple experimental methods to characterize monolayer mechanical properties are lacking. Here, we describe a system for tensile testing of freely suspended cultured monolayers that enables the examination of their mechanical behavior at multi-, uni-, and subcellular scales. Using this system, we provide measurements of monolayer elasticity and show that this is two orders of magnitude larger than the elasticity of their isolated cellular components. Monolayers could withstand more than a doubling in length before failing through rupture of intercellular junctions. Measurement of stress at fracture enabled a first estimation of the average force needed to separate cells within truly mature monolayers, approximately ninefold larger than measured in pairs of isolated cells. As in single cells, monolayer mechanical properties were strongly dependent on the integrity of the actin cytoskeleton, myosin, and intercellular adhesions interfacing adjacent cells. High magnification imaging revealed that keratin filaments became progressively stretched during extension, suggesting they participate in monolayer mechanics. This multiscale study of monolayer response to deformation enabled by our device provides the first quantitative investigation of the link between monolayer biology and mechanics.

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This paper presents an adaptive Sequential Monte Carlo approach for real-time applications. Sequential Monte Carlo method is employed to estimate the states of dynamic systems using weighted particles. The proposed approach reduces the run-time computation complexity by adapting the size of the particle set. Multiple processing elements on FPGAs are dynamically allocated for improved energy efficiency without violating real-time constraints. A robot localisation application is developed based on the proposed approach. Compared to a non-adaptive implementation, the dynamic energy consumption is reduced by up to 70% without affecting the quality of solutions. © 2012 IEEE.

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Optical motion capture systems suffer from marker occlusions resulting in loss of useful information. This paper addresses the problem of real-time joint localisation of legged skeletons in the presence of such missing data. The data is assumed to be labelled 3d marker positions from a motion capture system. An integrated framework is presented which predicts the occluded marker positions using a Variable Turn Model within an Unscented Kalman filter. Inferred information from neighbouring markers is used as observation states; these constraints are efficient, simple, and real-time implementable. This work also takes advantage of the common case that missing markers are still visible to a single camera, by combining predictions with under-determined positions, resulting in more accurate predictions. An Inverse Kinematics technique is then applied ensuring that the bone lengths remain constant over time; the system can thereby maintain a continuous data-flow. The marker and Centre of Rotation (CoR) positions can be calculated with high accuracy even in cases where markers are occluded for a long period of time. Our methodology is tested against some of the most popular methods for marker prediction and the results confirm that our approach outperforms these methods in estimating both marker and CoR positions. © 2012 Springer-Verlag.

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This paper presents a heterogeneous reconfigurable system for real-time applications applying particle filters. The system consists of an FPGA and a multi-threaded CPU. We propose a method to adapt the number of particles dynamically and utilise the run-time reconfigurability of the FPGA for reduced power and energy consumption. An application is developed which involves simultaneous mobile robot localisation and people tracking. It shows that the proposed adaptive particle filter can reduce up to 99% of computation time. Using run-time reconfiguration, we achieve 34% reduction in idle power and save 26-34% of system energy. Our proposed system is up to 7.39 times faster and 3.65 times more energy efficient than the Intel Xeon X5650 CPU with 12 threads, and 1.3 times faster and 2.13 times more energy efficient than an NVIDIA Tesla C2070 GPU. © 2013 Springer-Verlag.

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Detecting receptor dimerisation and other forms of clustering on the cell surface depends on methods capable of determining protein-protein separations with high resolution in the ∼10-50 nm range. However, this distance range poses a significant challenge because it is too large for fluorescence resonance energy transfer and contains distances too small for all other techniques capable of high-resolution in cells. Here we have adapted the technique of fluorophore localisation imaging with photobleaching to measure inter-receptor separations in the cellular environment. Using the epidermal growth factor receptor, a key cancer target molecule, we demonstrate ∼10 nm resolution while continuously covering the range of ∼10-80 nm. By labelling the receptor on cells expressing low receptor numbers with a fluorescent antagonist we have found inter-receptor separations all the way up from 8 nm to 59 nm. Our data are consistent with epidermal growth factor receptors being able to form homo-polymers of at least 10 receptors in the absence of activating ligands. © 2013 Needham et al.

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Cell monolayers line most of the surfaces and cavities in the human body. During development and normal physiology, monolayers sustain, detect and generate mechanical stresses, yet little is known about their mechanical properties. We describe a cell culture and mechanical testing protocol for generating freely suspended cell monolayers and examining their mechanical and biological response to uniaxial stretch. Cells are cultured on temporary collagen scaffolds polymerized between two parallel glass capillaries. Once cells form a monolayer covering the collagen and the capillaries, the scaffold is removed with collagenase, leaving the monolayer suspended between the test rods. The suspended monolayers are subjected to stretching by prying the capillaries apart with a micromanipulator. The applied force can be measured for the characterization of monolayer mechanics. Monolayers can be imaged with standard optical microscopy to examine changes in cell morphology and subcellular organization concomitant with stretch. The entire preparation and testing protocol requires 3-4 d.

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We demonstrate a new method for extracting high-level scene information from the type of data available from simultaneous localisation and mapping systems. We model the scene with a collection of primitives (such as bounded planes), and make explicit use of both visible and occluded points in order to refine the model. Since our formulation allows for different kinds of primitives and an arbitrary number of each, we use Bayesian model evidence to compare very different models on an even footing. Additionally, by making use of Bayesian techniques we can also avoid explicitly finding the optimal assignment of map landmarks to primitives. The results show that explicit reasoning about occlusion improves model accuracy and yields models which are suitable for aiding data association. © 2011. The copyright of this document resides with its authors.