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Biomedical subjects

G Fuhr

Publications and source records attributed to G Fuhr.

30 records · Page 2Linked to original sources

Characterisation of cell movement by impedance measurement on fibroblasts grown on perforated Si-membranes.

Mouse fibroblasts grown on perforated Si-membranes (pore diameter approximately 10 microns have been studied to clarify cell locomotive ability. The cell motility was microscopically monitored by a time-lapse video system and, simultaneously, the impedance of the growing cells was measured every 5 s. The correlations between observed cell activities and measured impedance events are discussed and classified. The method is sensitive and allows discrimination between signals arising from translocation of single cells and those arising from filopodia activities. Both cell and filopodia motion could be detected. Designs of microdevices fabricated in semiconductor technology are presented.

3T3 Cells↗

Cell manipulation and cultivation under a.c. electric field influence in highly conductive culture media.

Extreme miniaturisation of electrodes enabled us to apply high-frequency electric fields (between 100 kHz and several hundred MHz) of field strengths up to 50 kV/m into cell suspensions of high conductivity (several S/m), such as original cell culture media. The active electrode areas were additionally decreased and modified by insulating the terminals and/or coating of the electrodes with thin dielectric layers. Micro scaled electrode structures were fabricated on glass or silicon wafers in semiconductor technology. It could theoretically and experimentally be shown that cells exhibit exclusively negative dielectrophoresis if suspended in highly conductive media. Therefore, they can be repulsed from surfaces by appropriate arrangements of electrodes and easily be manipulated in free solution. Adherently growing animal cells, like mouse fibroblasts (3T3, L929), were cultivated in Dulbecco's Modification of Eagle's Medium (DMEM) or RPMI 1640 under permanent field application (frequency: 10 MHz, field strength: 50-100 kV/m).

3T3 Cells↗

Radio-frequency microtools for particle and liver cell manipulation.

Single particles can be manipulated by applying high frequencies to ultramicro electrode arrays fabricated on planar structures. Heat production can be reduced to the extent that intense electric fields can be applied even to unmodified cell culture media. Animal cells grow normally in the high field (up to 100 kV/m) between such continuously energized multielectrodes. As with laser tweezers [1-3], this technique can capture particles and cells in field traps, generate linear movement, and permit cell cultivation. It can also produce micropatterns of pH gradients, field-cast objects, and control cell adhesion. These microtools may be combined to develop cell separators, microsensors, and controlled-biocompatibility surfaces.

Animals↗

Three-dimensional electric field traps for manipulation of cells--calculation and experimental verification.

The forces acting on dielectric particles and living cells exposed to alternating and rotating fields generated by three-dimensional multi-electrode arrangements are investigated. Numerical procedures are described for the calculation of the electric field distribution and forces. The physical treatment considers electrodes of any shape and dielectric particles of complex structure. Particle and cell trapping are based on negative dielectrophoretic forces produced by high-frequency a.c. or rotating electric fields up to 400 MHz. Various multi-electrode systems were realised in commercially fabricated microelectrode systems, and tested for their ability to move and assemble microparticles or living cells without contact with the electrodes. The field distribution and accuracy of phase-controlled power application was tested using individual artificial particles trapped in the electric field cage. Position and trajectories of particle motion were measured. The paper gives an overview of electrode and field cage design in the microscale range.

Animals↗

A traveling-wave micropump for aqueous solutions: comparison of 1 g and microgram results.

The operation of a micro-chip-based fluid circulator based on the interaction of a high-frequency traveling wave with a thermally generated inhomogeneity in an aqueous medium is investigated. The profiles of the electric field, the thermal gradient and the driving force within the device are derived numerically, and the dependence upon applied voltage under different experimental conditions is derived. The importance of convection in the operation of the device is assessed by operating it in various orientations, as well as under microgravity, and also by investigating the reversal and turn-on characteristics. The pump showed the unexpected ability to trap microparticles present in the driven fluid.

Electricity↗

Levitation, holding, and rotation of cells within traps made by high-frequency fields.

Biological cells and other particles can be electrically manipulated by means of negative dielectrophoresis within microchambers whose electrode geometry is of the order of the cell size. Very-high-frequency fields (50 MHz and above) and media of increased relative permittivity are especially suitable for the purpose, as shown by experimental data on levitation and rotation. It appears to be possible to move and rotate cells or particles at will using this technology.

Cytological Techniques↗

Steady state electrolysis and isoelectric focusing.

The properties of pH gradients formed by stationary electrolysis of weak mobile or fixed electrolytes are analyzed. The model uses the appropriate balance equations and those of chemical equilibria. It is shown how the equation of the current density has to be modified for considering that fraction of current that is associated with the diffusion of neutral buffer molecules within a pH gradient. Furthermore it is shown that the pH gradients themselves give rise to water production within the gradient and that essential properties of the steady state are related to chemical reactions between the electrolyte constituents. The differential equations describing the gradients of the concentration of a given component, the pH, conductivity and potential are explicitly formulated in relation to those reactions. The equations are solved numerically and the significance of the results for isoelectric focusing is discussed. The experimental conditions to reach shallow and smooth pH gradients exhibiting sufficient ionic strength are formulated.

Buffers↗

Reversible and irreversible rotating field-induced membrane modifications.

We analyse the charge distribution as submitted by additionally induced transmembrane potentials in rotating electric fields. In contrast to d.c. and a.c. fields, in rotating fields the induced peak potential differences across the membrane systems are phase shifted with respect both to each other and to the external field vector. These phase differences are strongly frequency-dependent but were also influenced by the electrical properties of both the cell and the surrounding medium. We extend our investigation up to the non-linear field strength range of electrorotation and found reversible and irreversible changes in the rotational behavior of several cells. The most convenient variables for describing non-linear electrorotation are the characteristic frequency (fc1) and the corresponding angular velocity (omega c) of the cells. With increasing field strength the observed rotational behavior becomes more and more irreversible and finally rupture of the membrane occurs.

Animals↗

Differences in the rotation spectra of mouse oocytes and zygotes.

Rotation spectra of mouse oocytes, zygotes and embryos in the two-cell stage under the influence of high-frequency rotating fields were studied. The characteristic frequency (fc1) of cells isolated from superovulated + mated mice is different from that of oocytes. This was attributed to an increase in the membrane resistance and, less probably, to a change in the zona pellucida conductivity. The rotation spectra can be used to differentiate between non-fertilized and fertilized eggs. A theoretical interpretation of the measured spectra and simulation of the changes caused by fertilization is given.

Animals↗

Rotation of dielectrics in a rotating electric high-frequency field. Model experiments and theoretical explanation of the rotation effect of living cells.

Model experiments are carried out to clarify the mechanism of rotation of living cells in a rotating electric field. According to classical investigations of the rotation of macroscopic bodies in external fields, the rotation of spherical glass vessels or metal cylinder filled with electrolyte solutions was investigated. The relation of the calculations of Lertes (1921a,b) to the recent paper of Arnold and Zimmerman (1982) and our new derivations lead to equations explaining the rotation of objects. The results are compared with measurements using mesophyll protoplasts and data from the literature.

Cell Movement↗

Traveling-wave dielectrophoresis of microparticles.

The traveling-wave-induced linear transfer of dielectric particles like living cells and artificial objects of microscopic dimensions is analyzed. It is shown that the electrode geometries must correspond to particle sizes to allow an effective manipulation of particles immersed in weakly electrolytic solutions by high frequency traveling waves. The theoretical model elaborated in this paper is in good agreement with experimental results obtained in microfabricated chambers of linearly arranged electrodes. It explains the behavior of homogeneous cellulose spheres as well as that of membrane-covered pine polls. The traveling-wave-driven electrodes are described by a superposition of time-dependent point charges. Subsequently, each of these point charges has to be considered as polarizing the dielectric particle and interacting with the polarized particle. This results in forces which effectively translocate the particle.

Cellulose↗

Cell traces--footprints of individual cells during locomotion and adhesion.

Animal cells release traces of material onto glass or silicon surfaces during adhesion and migration. This little studied phenomenon is a widespread and normal concomitant of cell migration. The paper introduces the study of such material. The traces can be visualised by different microscopic techniques (e.g. TIRF, IRM, CLSM, AFM, SEM). Cell traces typical for different cell lines (NIH 3T3 and L929 mouse fibroblasts, mouse macrophages, mouse sarcoma cells and human osteosarcoma cells) are shown and discussed. There are well organised structures such as different linear and nodular elements as well as patches. Traces can extend up to some hundred micrometers from the cell, but the dimensions of the linear elements are in the submicron range. Cell traces are not identical with focal contacts but can include them. A first classification of basic elements is proposed. It allows an estimation of the total volume and surface in comparison to the donor cell. Higher order structures are discussed and a first insight into the protein composition of traces produced by mouse fibroblasts is given. Our observations, together with the cell adhesion literature suggest that the amount of released material, its extent and chemical and structural properties depend on cell type and physiology as well as other external influences. Cell traces in combination with the adhesion pattern of the donor cell should give information about the activity and physiological status of individual cells, the mechanisms of cell locomotion and the molecular composition of the donor cell membrane. The traces might possibly be used as submicron elements for passive electric characterisation and biotechnological applications.

Animals↗