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Biomedical subjects

G H Markx

Publications and source records attributed to G H Markx.

8 recordsLinked to original sources

Dielectrophoresis of reverse phase emulsions.

Reverse miniemulsions, emulsions of droplets of size 200 nm-1 microm of a polar liquid dispersed in an apolar continuous liquid phase, exhibit strong electrokinetic responses in low-frequency electric fields. The electrokinetic behaviour of a reverse miniemulsion, previously developed for use as electronic paper, has been investigated under static and flow conditions, in uniform and non-uniform electric fields. Results reveal that when using frequencies lower than 10 Hz strong aggregation of the droplets occurs. In uniform electric fields, under static conditions, droplets reversibly aggregate into honeycomb-like or irregular aggregates. Under flow conditions, droplets aggregate into approximately equidistant streams. In non-uniform electric fields the droplets reversibly aggregate in high-field regions, and can be guided along regions of high field strength in a flow. The potential of the technique for the formation of structured materials is discussed.

Cell Separation↗

Applications of dielectrophoresis in biotechnology.

Recent progress in the development of microelectrode structures has led to new techniques for the dielectrophoretic characterization and sorting of cells, microorganisms and other bioparticles using nonuniform AC electric fields. These methods utilize differences in the dielectric polarizabilities of cells for their effectiveness, and factors controlling such properties include the conductivity and permittivity of membranes and any cell walls, electrical double layers associated with surface charges, cell morphologies, and internal structures. Applications of dielectrophoresis have included the selective spatial manipulation and separation of mixtures of bacteria, viable and unviable cells, cancerous and normal cells, and red and white blood cells.

Animals↗

Dielectrophoretic separation of bacteria using a conductivity gradient.

Dielectrophoresis, the lateral motion induced on particles by non-uniform electric fields, is a sensitive function of the electrical conductivity of the particle suspending medium. This dependence is exploited in a new technique for separating bioparticles from suspended mixtures. The bioparticles are first immobilised by positive dielectrophoresis at electrodes in a separation chamber, and the conductivity of the liquid flowing through the chamber is then gradually and continuously increased so as to produce a conductivity gradient with time. The bioparticles are released from the electrodes according to their own dielectric properties and as a function of flow rate and medium conductivity. This is demonstrated for pure suspensions and mixtures of the bacteria Bacillus subtilis, Escherichia coli and Micrococcus luteus.

Bacillus subtilis↗

Effect of biocide concentration on electrorotation spectra of yeast cells.

The effect of the biocide Cosmocil (polyhexanide) at different concentrations on the electrorotation spectra of yeast cells is investigated over the frequency range from 1 kHz to 10 MHz. The dielectric properties of the yeast, before and after biocide treatment, were deduced from the electrorotation spectra using two-shell ellipsoid modelling methods that have been well tested for other heterogeneous biological systems. The results show a gradual increase in the cytoplasmic membrane conductivity with increasing biocide concentration, rather than an "all-or-nothing' breakdown of the membrane. The technique gives a quantitative analysis of the toxic damage by chemicals to cells and can be exploited in the development of new pharmacological agents.

Biguanides↗

Differentiation of viable and non-viable bacterial biofilms using electrorotation.

A new technique for studying the properties of biofilms has been developed, based on the phenomenon of electrorotation. Biofilms of Klebsiella rubiacearum were formed on the surfaces of 6 microns diameter polystyrene beads, and the presence of such films was found to alter their electrorotation spectra. The effects of adding a biocide (polyhexanide) to the surrounding aqueous medium was also investigated. The dielectric properties of the beads with biofilms, before and after biocide treatment, were interpreted from the electrorotation spectra using modelling methods that have been well tested for other heterogeneous biological systems. The technique is of value in understanding the physico-chemical properties of biofilms and can be adapted for monitoring the presence of toxic chemicals and for testing the activity of biocides against biofilms.

Biofilms↗

Separation of viable and non-viable yeast using dielectrophoresis.

Dielectrophoresis, the movement of particles in non-uniform AC electric fields, was used to rapidly separate viable and non-viable yeast cells with good efficiency. Known mixtures of viable and heat-treated cells of Saccharomyces cerevisiae were separated and selectively isolated using positive and negative dielectrophoretic forces generated by microelectrodes in a small chamber. Good correlations with the initial known relative compositions were obtained by direct microscopic counting of cells at the electrodes after initial dielectrophoretic separation (r = 0.995), from methylene blue staining (r = 0.992) and by optical absorption measurements (r = 0.980) of the effluent after selectively flushing out the viable and non-viable cells from the chamber. Through measurement of cell viability by staining with methylene blue and plate counts, for an initial suspension of approx. 1.4 x 10(7) cells per ml containing 60% non-viable cells, the dielectrophoretically separated non-viable fraction contained 3% viable cells and the viable fraction 8% dead cells. The separation efficiency is increased by dilution of the initial suspension or by repeat operation(s). Cell viability was not affected by the separation procedure.

Electrophoresis↗

Dielectric spectroscopy as a novel and convenient tool for the study of the shear sensitivity of plant cells in suspension culture.

Plant cell suspensions of different species and different age were subjected to hydrodynamic stress while following the decline in the volume fraction of intact cells by measuring the permittivity of the cell suspension at radio frequencies. Results were compared with the fresh weight, dry weight, packed cell volume and cell number of the suspensions. At first a rapid decline is seen as the most shear-sensitive cells are broken up, followed by a slower decline as less sensitive cells are broken up. The sensitivity of the cells to shear stress depended strongly on the cell line used but only slightly on their age, older cells being more sensitive. The dependence of the shear sensitivity on the cell line might be an effect of the species investigated, the culturing conditions of the cell line, or both. It was found that cells that grow in a finely dispersed suspension are much less prone to shear stress than is often assumed.

Cells, Cultured↗

The use of dielectric permittivity for the control of the biomass level during biotransformations of toxic substrates in continuous culture.

Since the permittivity signal of a cell suspension measured using dielectric spectroscopy at radio frequencies is essentially determined only by viable (intact) cells, it can be used to monitor the concentration of viable cells in a fermentor in which a large proportion of the cells is nonviable. This could be used to select for organisms that are highly resistant to stress, for example from toxic chemicals used in biotransformations. We sought to control the concentration of viable yeast cells in a fermentor by adding small amounts of benzaldehyde, thus imposing a selection regime for cells highly resistant to benzaldehyde. However, after the addition of benzaldehyde, an increase in the permittivity is seen first followed by a decrease, thus making the control of biomass using a standard on-off controller difficult. It is shown that it is possible effectively to control the level of viable biomass in the fermentor in the presence of a large concentration of necromass using a combination of an inverse response compensator and a PID controller.

Algorithms↗