Functional gold nanoparticle-peptide complexes as cell-targeting agents

In this paper, we report a novel approach using peptide CALNN and its derivative CALNNGGRRRRRRRR (CALNNR(8)) to functionalize gold nanoparticles for intracellular component targeting. The translocation is effected by the nanoparticle diameter and CALNNR8 surface coverage. The intracellular distributions of the complexes are change from the cellular nucleus to the endoplasmic reticulum by increasing the density of CALNNR8 at a constant nanoparticle diameter. Additionally, increasing the nanoparticle diameter at a constant density of CALNNR8 leads to less cellular internalization.

Gold nanoparticle-based near-infrared fluorescent detection of biological thiols in human plasma

In this work, a new fluorescent method for sensitive detection of biological thiols in human plasma was developed using a near-infrared (NIR) fluorescent dye, FR 730. The sensing approach was based on the strong affinity of thiols to gold and highly efficient fluorescent quenching ability of gold nanoparticles (Au NPs). In the presence of thiols, the NIR fluorescence would enhance dramatically due to desorption of FR 730 from the surfaces of Au NPs, which allowed the analysis of thiol-containing amino acids in a very simple approach. The size of Au NPs was found to affect the fluorescent assay and the best response for cysteine detection was achieved when using Au NPs with the diameter of 24 nm, where a linear range of 2.5 x 10(-8) M to 4.0 x 10(-6) M and a detection limit of as low as 10 nM was obtained. This method also demonstrated a high selectivity to thiol-containing amino acids due to the strong affinity of thiols to gold.

Electrochemical detection of DNA immobilized on gold colloid particles modified self-assembled monolayer electrode with silver nanoparticle label

The target DNA was immobilized successfully on gold colloid particles associated with a cysteamine monolayer on gold electrode surface. Self-assembly of colloidal An onto a cysteamine modified gold electrode can enlarge the electrode surface area and enhance greatly the amount of immobilized single stranded DNA (ssDNA). The electrontransfer processes of [Fe(CN)(6)](4)-/[Fe(CN)(6)](3-) on the gold surface were blocked due to the procedures of the target DNA immobilization, which was investigated by impedance spectroscopy. Then single stranded target DNA immobilized on the gold electrode hybridized with the silver nanoparticle-oligonucleotide DNA probe, followed by the release of the silver metal atoms anchored on the hybrids by oxidative metal dissolution, and the indirect determination of the released solubilized Ag-1 ions by anodic stripping voltammetry (ASV) at a carbon fiber microelectrode. The results show that this method has good correlation for DNA detection in the range of 10-800 pmol/1 and allows the detection level as low as 5 pmol/1 of the target oligonucleotides.

Head Tracking via Robust Registration in Texture Map Images

A novel method for 3D head tracking in the presence of large head rotations and facial expression changes is described. Tracking is formulated in terms of color image registration in the texture map of a 3D surface model. Model appearance is recursively updated via image mosaicking in the texture map as the head orientation varies. The resulting dynamic texture map provides a stabilized view of the face that can be used as input to many existing 2D techniques for face recognition, facial expressions analysis, lip reading, and eye tracking. Parameters are estimated via a robust minimization procedure; this provides robustness to occlusions, wrinkles, shadows, and specular highlights. The system was tested on a variety of sequences taken with low quality, uncalibrated video cameras. Experimental results are reported.

Tracking a Large Number of Objects from Multiple Views

We propose a multi-object multi-camera framework for tracking large numbers of tightly-spaced objects that rapidly move in three dimensions. We formulate the problem of finding correspondences across multiple views as a multidimensional assignment problem and use a greedy randomized adaptive search procedure to solve this NP-hard problem efficiently. To account for occlusions, we relax the one-to-one constraint that one measurement corresponds to one object and iteratively solve the relaxed assignment problem. After correspondences are established, object trajectories are estimated by stereoscopic reconstruction using an epipolar-neighborhood search. We embedded our method into a tracker-to-tracker multi-view fusion system that not only obtains the three-dimensional trajectories of closely-moving objects but also accurately settles track uncertainties that could not be resolved from single views due to occlusion. We conducted experiments to validate our greedy assignment procedure and our technique to recover from occlusions. We successfully track hundreds of flying bats and provide an analysis of their group behavior based on 150 reconstructed 3D trajectories.

Fast, Reliable Head Tracking under Varying Illumination: An Approach Based on Registration of Texture-Mapped 3D Model

An improved technique for 3D head tracking under varying illumination conditions is proposed. The head is modeled as a texture mapped cylinder. Tracking is formulated as an image registration problem in the cylinder's texture map image. The resulting dynamic texture map provides a stabilized view of the face that can be used as input to many existing 2D techniques for face recognition, facial expressions analysis, lip reading, and eye tracking. To solve the registration problem in the presence of lighting variation and head motion, the residual error of registration is modeled as a linear combination of texture warping templates and orthogonal illumination templates. Fast and stable on-line tracking is achieved via regularized, weighted least squares minimization of the registration error. The regularization term tends to limit potential ambiguities that arise in the warping and illumination templates. It enables stable tracking over extended sequences. Tracking does not require a precise initial fit of the model; the system is initialized automatically using a simple 2D face detector. The only assumption is that the target is facing the camera in the first frame of the sequence. The formulation is tailored to take advantage of texture mapping hardware available in many workstations, PC's, and game consoles. The non-optimized implementation runs at about 15 frames per second on a SGI O2 graphic workstation. Extensive experiments evaluating the effectiveness of the formulation are reported. The sensitivity of the technique to illumination, regularization parameters, errors in the initial positioning and internal camera parameters are analyzed. Examples and applications of tracking are reported.

Modular average case analysis: Language implementation and extension

Motivated by accurate average-case analysis, MOdular Quantitative Analysis (MOQA) is developed at the Centre for Efficiency Oriented Languages (CEOL). In essence, MOQA allows the programmer to determine the average running time of a broad class of programmes directly from the code in a (semi-)automated way. The MOQA approach has the property of randomness preservation which means that applying any operation to a random structure, results in an output isomorphic to one or more random structures, which is key to systematic timing. Based on original MOQA research, we discuss the design and implementation of a new domain specific scripting language based on randomness preserving operations and random structures. It is designed to facilitate compositional timing by systematically tracking the distributions of inputs and outputs. The notion of a labelled partial order (LPO) is the basic data type in the language. The programmer uses built-in MOQA operations together with restricted control flow statements to design MOQA programs. This MOQA language is formally specified both syntactically and semantically in this thesis. A practical language interpreter implementation is provided and discussed. By analysing new algorithms and data restructuring operations, we demonstrate the wide applicability of the MOQA approach. Also we extend MOQA theory to a number of other domains besides average-case analysis. We show the strong connection between MOQA and parallel computing, reversible computing and data entropy analysis.

Integrated genetic analysis systems

The overall objective of this thesis is to integrate a number of micro/nanotechnologies into integrated cartridge type systems to implement such biochemical protocols. Instrumentation and systems were developed to interface such cartridge systems: (i) implementing microfluidic handling, (ii) executing thermal control during biochemical protocols and (iii) detection of biomolecules associated with inherited or infectious disease. This system implements biochemical protocols for DNA extraction, amplification and detection. A digital microfluidic chip (ElectroWetting on Dielectric) manipulated droplets of sample and reagent implementing sample preparation protocols. The cartridge system also integrated a planar magnetic microcoil device to generate local magnetic field gradients, manipulating magnetic beads. For hybridisation detection a fluorescence microarray, screening for mutations associated with CFTR gene is printed on a waveguide surface and integrated within the cartridge. A second cartridge system was developed to implement amplification and detection screening for DNA associated with disease-causing pathogens e.g. Escherichia coli. This system incorporates (i) elastomeric pinch valves isolating liquids during biochemical protocols and (ii) a silver nanoparticle microarray for fluorescent signal enhancement, using localized surface plasmon resonance. The microfluidic structures facilitated the sample and reagent to be loaded and moved between chambers with external heaters implementing thermal steps for nucleic acid amplification and detection. In a technique allowing probe DNA to be immobilised within a microfluidic system using (3D) hydrogel structures a prepolymer solution containing probe DNA was formulated and introduced into the microfluidic channel. Photo-polymerisation was undertaken forming 3D hydrogel structures attached to the microfluidic channel surface. The prepolymer material, poly-ethyleneglycol (PEG), was used to form hydrogel structures containing probe DNA. This hydrogel formulation process was fast compared to conventional biomolecule immobilization techniques and was also biocompatible with the immobilised biomolecules, as verified by on-chip hybridisation assays. This process allowed control over hydrogel height growth at the micron scale.

Deposition of silver nanoparticles in geochemically heterogeneous porous media: predicting affinity from surface composition analysis.

The transport of uncoated silver nanoparticles (AgNPs) in a porous medium composed of silica glass beads modified with a partial coverage of iron oxide (hematite) was studied and compared to that in a porous medium composed of unmodified glass beads (GB). At a pH lower than the point of zero charge (PZC) of hematite, the affinity of AgNPs for a hematite-coated glass bead (FeO-GB) surface was significantly higher than that for an uncoated surface. There was a linear correlation between the average nanoparticle affinity for media composed of mixtures of FeO-GB and GB collectors and the relative composition of those media as quantified by the attachment efficiency over a range of mixing mass ratios of the two types of collectors, so that the average AgNPs affinity for these media is readily predicted from the mass (or surface) weighted average of affinities for each of the surface types. X-ray photoelectron spectroscopy (XPS) was used to quantify the composition of the collector surface as a basis for predicting the affinity between the nanoparticles for a heterogeneous collector surface. A correlation was also observed between the local abundances of AgNPs and FeO on the collector surface.

Far-field analysis of axially symmetric three-dimensional directional cloaks.

Axisymmetric radiating and scattering structures whose rotational invariance is broken by non-axisymmetric excitations present an important class of problems in electromagnetics. For such problems, a cylindrical wave decomposition formalism can be used to efficiently obtain numerical solutions to the full-wave frequency-domain problem. Often, the far-field, or Fraunhofer region is of particular interest in scattering cross-section and radiation pattern calculations; yet, it is usually impractical to compute full-wave solutions for this region. Here, we propose a generalization of the Stratton-Chu far-field integral adapted for 2.5D formalism. The integration over a closed, axially symmetric surface is analytically reduced to a line integral on a meridional plane. We benchmark this computational technique by comparing it with analytical Mie solutions for a plasmonic nanoparticle, and apply it to the design of a three-dimensional polarization-insensitive cloak.

An analysis of the efficacy of low-scale concentrated photovoltaics

Gemstone Team LEAF (Light Energy Acquisition of the Future)

Long-duration animal tracking in difficult lighting conditions.

High-throughput analysis of animal behavior requires software to analyze videos. Such software typically depends on the experiments' being performed in good lighting conditions, but this ideal is difficult or impossible to achieve for certain classes of experiments. Here, we describe techniques that allow long-duration positional tracking in difficult lighting conditions with strong shadows or recurring "on"/"off" changes in lighting. The latter condition will likely become increasingly common, e.g., for Drosophila due to the advent of red-shifted channel rhodopsins. The techniques enabled tracking with good accuracy in three types of experiments with difficult lighting conditions in our lab. Our technique handling shadows relies on single-animal tracking and on shadows' and flies' being accurately distinguishable by distance to the center of the arena (or a similar geometric rule); the other techniques should be broadly applicable. We implemented the techniques as extensions of the widely-used tracking software Ctrax; however, they are relatively simple, not specific to Drosophila, and could be added to other trackers as well.

Exploring the benefits of using multi-touch devices in performing information search tasks: An analysis in terms of performance, cognitive load and task engagement

Over the last decade, multi-touch devices (MTD) have spread in a range of contexts. In the learning context, MTD accessibility leads more and more teachers to use them in their classroom, assuming that it will improve the learning activities. Despite a growing interest, only few studies have focused on the impacts of MTD use in terms of performance and suitability in a learning context.However, even if the use of touch-sensitive screens rather than a mouse and keyboard seems to be the easiest and fastest way to realize common learning tasks (as for instance web surfing), we notice that the use of MTD may lead to a less favorable outcome. More precisely, tasks that require users to generate complex and/or less common gestures may increase extrinsic cognitive load and impair performance, especially for intrinsically complex tasks. It is hypothesized that task and gesture complexity will affect users’ cognitive resources and decrease task efficacy and efficiency. Because MTD are supposed to be more appealing, it is assumed that it will also impact cognitive absorption. The present study also takes into account user’s prior knowledge concerning MTD use and gestures by using experience with MTD as a moderator. Sixty university students were asked to perform information search tasks on an online encyclopedia. Tasks were set up so that users had to generate the most commonly used mouse actions (e.g. left/right click, scrolling, zooming, text encoding…). Two conditions were created: MTD use and laptop use (with mouse and keyboard) in order to make a comparison between the two devices. An eye tracking device was used to measure user’s attention and cognitive load. Our study sheds light on some important aspects towards the use of MTD and the added value compared to a laptop in a student learning context.

Exploring eye movement analysis as a measure of selective visual attention in brain injured individuals

Primary Objective: To investigate the utility of using a new method of assessment for deficits in selective visual attention (SVA). Methods and Procedures: An independent groups design compared six participants with brain injuries with six participants from a non-brain injured control group. The Sensomotoric Instruments Eye Movement system with remote eye-tracking device (eye camera), and 2 sets of eight stimuli were employed to determine if the camera would be a sensitive discriminator of SVA in these groups. Main Outcomes and Results: The attention profile displayed by the brain injured group showed that they were slower, made more errors, were less accurate, and more indecisive than the control group. Conclusions: The utility of eye movement analysis as an assessment method was established, with implications for rehabilitation requiring further development. Key words: selective visual attention, eye movement analysis, brain injury