Electric field stimulation of bipolar cells in a degenerated retina--a theoretical study.

Subretinal implants are the subject of clinical investigation for their ability to evoke useful visual sensations in blind individuals via electrical stimulation of the diseased retina. We investigated the spatial characteristic of the retinal polarization obtained by electric field stimulation through a subretinally located monopolar electrode array and bipolar electrode array. By combining electric potential simulation through a boundary element method with a segmented cell model, we computed the membrane voltage at the axon terminal of the bipolar cells as a function of the axon length (50-110 microm) and the electrode diameter. We found that short OFF bipolar cells are predominantly addressed by small bipolar electrodes (diameter between 60 and 100 microm) and by using a short duration ( < 150 micros) of the stimulating voltage pulse. Long ON cells are best addressed by large monopolar electrodes (diameter > 100 microm) and a long pulse duration ( > 150 micros). However, the low selectivity of the electric field stimulation with regard to the cell length does not enable the individual depolarization of long OFF cells and short ON cells. When the stimulation must take place at multiple retinal sites simultaneously, the bipolar electrode arrays allow for higher spatial modulation of the polarization of the axon terminal than the monopolar arrays.

The surface energy of various biomaterials coated with adhesion molecules used in cell culture.

This study calculates the surface energy of polystyrene tissue culture plastic, silicon, silicon dioxide and indium tin oxide, all of which have applications in tissue culture. The adhesion molecules: collagen, fibronectin, poly-L-ornithine and poly-D-lysine, were coated onto these various surfaces, and the surface energy of the coated substrates calculated. Coating with fibronectin was found to produce a monopolar acidic surface while poly-D-lysine, poly-L-ornithine and collagen coatings were found to produce monopolar basic surfaces. The calculated surface energy components of the coated materials were then used to give a quantitative determination of the magnitude of their hydrophobicity. It was concluded that collagen, polylysine and polyornithine could provide a hydrophobic or hydrophilic surface depending on the underlying substrates they were coated on. The measurement obtained for fibronectin, unlike the other adhesion molecules, was independent of the underlying surface and remained hydrophobic on all substrates tested. Wetting experiments were carried out on the coated substrates, using the tissue culture medium Dulbeccos modified eagles medium, both containing and not containing serum proteins, and saline solution. These liquids that are commonly used in tissue culture, were then used to provide information how these liquids behave on various substrates coated with the adhesion molecules. Results show that fibronectin coated surfaces represent the most phobic surface for all three liquids. The findings of this study can be used in cell manipulation studies and provide a valuable data set for the biomedical and research industries.

Voltammetric characterisation of silicon-based microelectrode arrays and their application to mercury-free stripping voltammetry of copper ions.

This paper describes the electrochemical characterisation of a range of gold and platinum microelectrode arrays (MEAs) fabricated by standard photolithographic methods. The inter-electrode spacing, geometry, numbers and dimensions of the electrodes in the arrays were found to influence the voltammetric behaviours obtained. Excellent correlation was found between experimental data and theoretical predictions employing published models of microelectrode behaviour. Gold MEAs were evaluated for their applicability to copper determination in a soil extract sample, where agreement was found between the standard analytical method and a method based on underpotential deposition-anodic stripping voltammetry (UPD-ASV) at the MEAs, offering a mercury-free alternative for copper sensing.

Development of a respirometric biochip for embryo assessment.

A prototype respirometric biochip dedicated to monitoring oxygen consumption of preimplantation embryos has been developed. The biochip comprises a linear array of eight flow-through microchambers profiled on silicon substrate, and functions together with a phosphorescent oxygen sensitive probe and fluorescence plate reader detection. A high level of sensitivity to changes in dissolved oxygen was achieved through miniaturisation and optimization of biochip geometry, and incorporation of appropriate sealing and humidification systems. The biochips have allowed characterisation of oxygen consumption, by 2 cell and blastocyst stage preimplantation mouse embryos, through monitoring as few as ten preimplantation embryos over a one-hour time period. They provide a non-invasive, simple and convenient means for assessing preimplantation embryo metabolism.

A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing.

Cell viability assays represent an important technology in modern cell biology, drug discovery and biotechnology, where currently there is a high demand for simple, sensitive and cost-effective screening methods. We have developed a new methodology and associated tools for cell-based screening assays, which are based on the measurement of the rates of oxygen uptake in cells by luminescence quenching. Sealable microchamber devices matching the footprint of a standard 96-well plate were developed and used in conjunction with long-decay phosphorescent oxygen probes. These devices permit cell non-invasive, real-time monitoring of cellular respiration and a rapid, one-step, kinetic assessment of multiple samples for cell viability, drug/effector action. These assays can be carried out on conventional fluorescence plate readers, they are suitable for different types of cells, including adherent and slow-respiring cells, require small sample volumes and cell numbers, and are amenable for high throughput screening. Monitoring of as little as 300 mammalian cells in 3 microl volume has been demonstrated.

Application of magnetohydrodynamic actuation to continuous flow chemistry.

Continuous flow microreactors with an annular microchannel for cyclical chemical reactions were fabricated by either bulk micromachining in silicon or by rapid prototyping using EPON SU-8. Fluid propulsion in these unusual microchannels was achieved using AC magnetohydrodynamic (MHD) actuation. This integrated micropumping mechanism obviates the use of moving parts by acting locally on the electrolyte, exploiting its inherent conductive nature. Both silicon and SU-8 microreactors were capable of MHD actuation, attaining fluid velocities of the order of 300 microm s(-1) when using a 500 mM KCl electrolyte. The polymerase chain reaction (PCR), a thermocycling process, was chosen as an illustrative example of a cyclical chemistry. Accordingly, temperature zones were provided to enable a thermal cycle during each revolution. With this approach, fluid velocity determines cycle duration. Here, we report device fabrication and performance, a model to accurately describe fluid circulation by MHD actuation, and compatibility issues relating to this approach to chemistry.