A simple, sensitive and selective quantum-dot-based western blot method for the simultaneous detection of multiple targets from cell lysates

Quantum dots (Qdots) are fluorescent nanoparticles that have great potential as detection agents in biological applications. Their optical properties, including photostability and narrow, symmetrical emission bands with large Stokes shifts, and the potential for multiplexing of many different colours, give them significant advantages over traditionally used fluorescent dyes. Here, we report the straightforward generation of stable, covalent quantum dot-protein A/G bioconjugates that will be able to bind to almost any IgG antibody, and therefore can be used in many applications. An additional advantage is that the requirement for a secondary antibody is removed, simplifying experimental design. To demonstrate their use, we show their application in multiplexed western blotting. The sensitivity of Qdot conjugates is found to be superior to fluorescent dyes, and comparable to, or potentially better than, enhanced chemiluminescence. We show a true biological validation using a four-colour multiplexed western blot against a complex cell lysate background, and have significantly improved previously reported non-specific binding of the Qdots to cellular proteins.

A simple bioanalytical method for the quantification of antiepileptic drugs in dried blood spots

An increasing number of publications on the dried blood spot (DBS) sampling approach for the quantification of drugs and metabolites have been spurred on by the inherent advantages of this sampling technique. In the present research, a selective and sensitive high-performance liquid chromatography method for the concurrent determination of multiple antiepileptic drugs (AEDs) [levetiracetam (LVT), lamotrigine (LTG), phenobarbital (PHB)], carbamazepine (CBZ) and its active metabolite carbamazepine-10,11 epoxide (CBZE)] in a single DBS has been developed and validated. Whole blood was spotted onto Guthrie cards and dried. Using a standard punch (6 mm diameter), a circular disc was punched from the card and extracted with methanol: acetonitrile (3:1, v/v) containing hexobarbital (Internal Standard) and sonicated prior to evaporation. The extract was then dissolved in water and vortex mixed before undergoing solid phase extraction using HLB cartridges. Chromatographic separation of the AEDs was achieved using Waters XBridge™ C18 column with a gradient system. The developed method was linear over the concentration ranges studied with r ≥ 0.995 for all compounds. The lower limits of quantification (LLOQs) were 2, 1, 2, 0.5 and 1 μg/mL for LVT, LTG, PHB, CBZE and CBZ, respectively. Accuracy (%RE) and precision (%CV) values for within and between day were <20% at the LLOQs and <15% at all other concentrations tested. This method was successfully applied to the analysis of the AEDs in DBS samples taken from children with epilepsy for the assessment of their adherence to prescribed treatments.

Simple design of FBG-based VSB filters for ultra-dense WDM transmission

A novel approach to ultra-narrow optical vestigial sideband (VSB) filtering based on a specially designed fibre Bragg grating (FBG) operating in transmission is proposed. This design can be easily implemented by writing FBG-based filters for each WDM channel before MUX. The specially designed filter profile allows to take full advantage of crosstalk suppression while minimising waveform distortion.

A simple remote sensing based information system for monitoring sites of conservation importance

Monitoring is essential for conservation of sites, but capacity to undertake it in the field is often limited. Data collected by remote sensing has been identified as a partial solution to this problem, and is becoming a feasible option, since increasing quantities of satellite data in particular are becoming available to conservationists. When suitably classified, satellite imagery can be used to delineate land cover types such as forest, and to identify any changes over time. However, the conservation community lacks (a) a simple tool appropriate to the needs for monitoring change in all types of land cover (e.g. not just forest), and (b) an easily accessible information system which allows for simple land cover change analysis and data sharing to reduce duplication of effort. To meet these needs, we developed a web-based information system which allows users to assess land cover dynamics in and around protected areas (or other sites of conservation importance) from multi-temporal medium resolution satellite imagery. The system is based around an open access toolbox that pre-processes and classifies Landsat-type imagery, and then allows users to interactively verify the classification. These data are then open for others to utilize through the online information system. We first explain imagery processing and data accessibility features, and then demonstrate the toolbox and the value of user verification using a case study on Nakuru National Park, Kenya. Monitoring and detection of disturbances can support implementation of effective protection, assist the work of park managers and conservation scientists, and thus contribute to conservation planning, priority assessment and potentially to meeting monitoring needs for Aichi target 11.

Exploration of a simple, low cost, micro gas turbine recuperator solution for a domestic combined heat and power unit

For micro gas turbines (MGT) of around 1 kW or less, a commercially suitable recuperator must be used to produce a thermal efficiency suitable for use in UK Domestic Combined Heat and Power (DCHP). This paper uses computational fluid dynamics (CFD) to investigate a recuperator design based on a helically coiled pipe-in-pipe heat exchanger which utilises industry standard stock materials and manufacturing techniques. A suitable mesh strategy was established by geometrically modelling separate boundary layer volumes to satisfy y + near wall conditions. A higher mesh density was then used to resolve the core flow. A coiled pipe-in-pipe recuperator solution for a 1 kW MGT DCHP unit was established within the volume envelope suitable for a domestic wall-hung boiler. Using a low MGT pressure ratio (necessitated by using a turbocharger oil cooled journal bearing platform) meant unit size was larger than anticipated. Raising MGT pressure ratio from 2.15 to 2.5 could significantly reduce recuperator volume. Dimensional reasoning confirmed the existence of optimum pipe diameter combinations for minimum pressure drop. Maximum heat exchanger effectiveness was achieved using an optimum or minimum pressure drop pipe combination with large pipe length as opposed to a large pressure drop pipe combination with shorter pipe length. © 2011 Elsevier Ltd. All rights reserved.