Martin Arundell

High-speed particle detection in a micro-Coulter counter with two-dimensional adjustable aperture

This article presents the fabrication and characterisation of a high-speed detection micro-Coulter counter with two-dimensional (2D) adjustable aperture and differential impedance detection. The developed device has been fabricated from biocompatible and transparent materials (polymer and glass) and uses the principle of hydrodynamic focusing in two dimensions. The use of a conductive solution for the sample flux and non-conductive solutions for the focalising fluxes provides an adjustable sample flow where particles are aligned and the resistive response concentrated, consequently enhancing the sensitivity and versatility of the device. High-speed counting of 20 microm polystyrene particles and 5 microm yeast cells with a rate of up to 1,000 particles/s has been demonstrated. Two-dimensional focusing conditions have been used in devices with physical cross-sectional areas of 180 microm x 65 microm and 100 microm x 43 microm, respectively, in which particles resulted undetectable in the absence of focusing. The 2D-focusing conditions have provided, in addition, increased detection sensitivity by a factor of 1.6 as compared to 1D-focusing conditions.

Synapse‐specific changes in serotonin signalling contribute to age‐related changes in the feeding behaviour of the pond snail, Lymnaea

This study utilised the pond snail, Lymnaea to examine the contribution that alterations in serotonergic signalling make to age‐related changes in feeding. Age‐related decreases in 5‐HIAA levels in feeding ganglia were positively correlated with a decrease in the number of sucrose‐evoked bites and negatively correlated with an increase in inter‐bite interval, implicating alterations in serotonergic signalling in the aged phenotype. Analysis of the serotonergic cerebral giant cell (CGC) input to the protraction motor neurone (B1) demonstrated that fluoxetine (10–100 nM) increased the amplitude/duration of the evoked EPSP in both young and middle aged but not in old neurones, suggesting an age‐related attenuation of the serotonin transporter. 5‐HT evoked a concentration‐dependent increase in the amplitude/duration of B1 EPSP, which was greater in old neurones compared to both young and middle aged. Conversely, the 5‐HT‐evoked depolarisation and conditional bursting of the swallow motor neurone (B4) were attenuated in old neurones, functions critical for a full feeding rhythm. The CGCs’ ability to excite B1 was blocked by cinanserin but not by methysergide. Conversely, the CGC to B4 connection was completely blocked by methysergide and only partially by cinanserin suggesting that age‐related changes may be receptor‐specific. In summary, synapse‐specific attenuation of the CGC‐B4 connection and enhancement of the CGC‐B1 connection would slow the swallow phase and maintain protraction, consistent with behavioural observations.

Effects of age on feeding behavior and chemosensory processing in the pond snail, Lymnaea stagnalis

This study used behavioral and electrophysiological techniques to examine age-related changes in the feeding behavior and chemosensory processing in the pond snail, Lymnaea stagnalis. Increasing age was associated with a 50% decrease in long-term food consumption. Analysis of short-term sucrose-evoked feeding bouts showed an age-related increase in the number of animals that failed to respond to the stimulus. Of the animals that did respond increasing age was associated with a decrease in the number of sucrose-evoked bites and a increase in the duration of the swallow phase. These changes were observed with both 0.01 and 0.05M sucrose stimuli but were not seen when 0.1M sucrose was used as the stimulus. Electrophysiological analysis of the chemosensory pathway in semi-intact lip-CNS preparations failed to demonstrate a significant change in the neuronal information entering the cerebral ganglia from the lips via the median lip nerve, but did demonstrate an age-related deficit in the neuronal output from the cerebral ganglia. This deficit was also dependent on the sucrose concentration and mirrored the concentration-dependent changes in feeding behavior. In summary, aging appeared to affect central but not peripheral processing of chemosensory information and suggests that this deficit contributes to the age-related changes in feeding behavior.

Changes in the properties of the modulatory cerebral giant cells contribute to aging in the feeding system of Lymnaea

This study examined whether electrophysiological changes in the endogenous properties and connectivity of the modulatory serotonergic cerebral giant cells (CGCs) contributed to the age-related changes in feeding behavior of the pond snail, Lymnaea. With increasing age there was a decrease in spontaneous CGC firing rates and decreased excitability of the CGCs to both chemosensory stimulation (0.05M sucrose applied to the lips) and direct intracellular current injection. These changes could be accounted for by a decrease in the input resistance of the neuron and an increase in the amplitude and the duration of the after-hyperpolarization. Decreases were also seen in the % of CGC pairs that were electrically coupled causing asynchronous firing. Together these changes would tend to reduce the ability of the CGCs to gate and control the frequency of the feeding behavior. Part of the ability of the CGCs to gate and frequency control the feeding network is to provide a background level of excitation to the feeding motor neurons. Recordings from B1 and B4 motor neurons showed an age-related hyperpolarization of the resting membrane potential consistent with a deficit in CGC function. Increases were seen in the strength of the evoked CGC-->B1 connection, however, this increase failed to compensate for the deficits in CGC excitability. In summary, age-related changes in the properties of the CGCs were consistent with them contributing to the age-related changes in feeding behavior seen in Lymnaea.

Electrokinetic DNA transport in 20nm-high nanoslits: Evidence for movement through a wall-adsorbed polymer nanogel

The electrokinetic transport behavior of λ‐DNA (48 kbp) in 20 nm‐high fused‐silica nanoslits in the presence of short‐chain PVP is investigated. Mobility and video data show a number of phenomena that are typical of DNA transport through gels or polymer solutions, thus indicative of rigid migration obstacles in the DNA pathway. Calculations show that a several nanometer thin layer of wall‐adsorbed PVP (‘nano‐gel’) can provide such a rigid obstacle matrix to the DNA. Such ultrathin wall‐adsorbed polymer layers represent a new type of matrix for electrokinetic DNA separation.

Microelectrode investigation of neuroneal ageing from a single identified neurone.

Microelectrode amperometry is uniquely suited for characterising the dynamics of neurotransmitter release, as it offers unparalleled spatial and temporal resolution. We have used carbon fibre microelectrodes to study release of the monoamine neurotransmitter serotonin (5-HT) and the gaseous transmitter nitric oxide (NO) in intact central nervous system of the water snail, Lymnaea stagnalis. Analysis of spontaneous vesicular release of 5-HT and depolarisation-induced release of NO reveals significant differences with ageing that may be associated with changes in protein structure and function.

The anti-bacterial effect of an electrochemical anti-fouling method intended for the protection of miniaturised oceanographic sensors

An electrochemical anti-fouling method, based upon the generation of chlorine from seawater, was applied to a proprietary design of Lab on a Chip conductivity, temperature and dissolved oxygen sensor. The method was evaluated using PCR after a six-week field trial in which it significantly reduced the burden of bacterial biofouling.

Design of a miniaturized electrochemical instrument for in-situ O2 monitoring

The authors are working toward the design of a device for the detection of oxygen, following a discrete and an integrated instrumentation implementation. The discrete electronics are also used for preliminary analysis, to confirm the validity of the conception of system, and its set-up would be used in the characterization of the integrated device, waiting for the chip fabrication. This paper presents the design of a small and portable potentiostat integrated with electrodes, which is cheap and miniaturized, which can be applied for on-site measurements for the simultaneous detection of O2 and temperature in water systems. As a first approach a discrete PCB has been designed based on commercial discrete electronics and specific oxygen sensors. Dissolved oxygen concentration (DO) is an important index of water quality and the ability to measure the oxygen concentration and temperature at different positions and depths would be an important attribute to environmental analysis. Especially, the objective is that the sensor and the electronics can be integrated in a single encapsulated device able to be submerged in environmental water systems and be able to make multiple measurements. For our proposed application a small and portable device is developed, where electronics and sensors are miniaturized and placed in close proximity to each other. This system would be based on the sensors and electronics, forming one module, and connected to a portable notebook to save and analyze the measurements on-line. The key electronics is defined by the potentiostat amplifier, used to fix the voltage between the Working (WE) and Reference (RE) electrodes following an input voltage (Vin). Vin is a triangular signal, programmed by a LabView© interface, which is also used to represent the CV transfers. To obtain a smaller and compact solution the potentiostat amplifier has also been integrated defining a full custom ASIC amplifier, which is in progress, looking for a point-of-care device. These circuits have been designed with a 0.13 μm technology from ST Microelectronics through the CMP-TIMA service.

Buzz off! An evaluation of ultrasonic acoustic vibration for the disruption of marine micro-organisms on sensor-housing materials.

Biofouling is a process of ecological succession which begins with the attachment and colonization of micro‐organisms to a submerged surface. For marine sensors and their housings, biofouling can be one of the principle limitations to long‐term deployment and reliability. Conventional antibiofouling strategies using biocides can be hazardous to the environment, and therefore alternative chemical‐free methods are preferred. In this study, custom‐made testing assemblies were used to evaluate ultrasonic vibration as an antibiofouling process for marine sensor‐housing materials over a 28‐day time course. Microbial biofouling was measured based on (i) surface coverage, using fluorescence microscopy and (ii) bacterial 16S rDNA gene copies, using Quantitative polymerase chain reaction (PCR). Ultrasonic vibrations (20 KHz, 200 ms pulses at 2‐s intervals; total power 16·08 W) significantly reduced the surface coverage on two plastics, poly(methyl methacrylate) and polyvinyl chloride (PVC) for up to 28 days. Bacterial gene copy number was similarly reduced, but the results were only statistically significant for PVC, which displayed the greatest overall resistance to biofouling, regardless of whether ultrasonic vibration was applied. Copper sheet, which has intrinsic biocidal properties was resistant to biofouling during the early stages of the experiment, but inhibited measurements made by PCR and generated inconsistent results later on.

Bridging two cultures: minimalistic networks prepared by microfluidic arraying, and open access compartments for electrophysiology

Microfabrication protocols are described for two compartmentalized neuron culture platforms which extend beyond the capabilities of conventional systems. The first involves a differential flow microfluidic circuit for arraying single neurons, along with protocols for in chip biomaterial patterning and the selective treatment of somata or outgrowth compartments. These minimalistic neuronal networks are ideal for spatially resolved research using rare and precious neuronal subtypes as well as parallelization for screening biochemical libraries. The second, open-access, system solves the micro-to-macro interface challenge to enable the insertion of micromanipulators for electrophysiology studies or localized perturbation using a microinjector. This system is especially useful for the spatiotemporal investigation of mechanisms underlying disease, such as neurodegeneration and epileptic seizures. Design files along with soft lithography replication techniques are provided to facilitate the straightforward uptake of these platforms. Technology integration approaches are also presented for the alignment of arrayed neurons with individual microelectrodes for highly parallel electrophysiological and electrochemical measurements throughout nodes in the compartmentalized neuronal network.