An electrode system is described for the near-simultaneous application and measurement of translational, levitational and rotational forces induced by AC electric fields, and this has been used to investigate the differences in the AC electrodynamics of viable and non-viable yeast cells. A new approach to the theoretical modelling of the experimental data has enabled these differences to be quantified in terms of changes in the conductivity of the cytoplasmic membrane and cell interior. The results are considered to have potentially important biomedical and biotechnological applications.
We have trapped single protein molecules of R-phycoerythrin in an aqueous solution by an alternating electric field. A radio frequency voltage is applied to sharp nanoelectrodes and hence produces a strong electric field gradient. The resulting dielectrophoretic forces attract freely diffusing protein molecules. Trapping takes place at the electrode tips. Switching off the field immediately releases the molecules. The electric field distribution is computed, and from this the dielectrophoretic response of the molecules is calculated using a standard polarization model. The resulting forces are compared to the impact of Brownian motion. Finally, we discuss the experimental observations on the basis of the model calculations.
The dielectrophoresis (DEP) data reported in the literature since 1994 for 22 different globular proteins is examined in detail. Apart from three cases, all of the reported protein DEP experiments employed a gradient field factor ∇ E m 2 that is much smaller (in some instances by many orders of magnitude) than the ~4 × 1021 V2/m3 required, according to current DEP theory, to overcome the dispersive forces associated with Brownian motion. This failing results from the macroscopic Clausius–Mossotti (CM) factor being restricted to the range 1.0 > CM > −0.5. Current DEP theory precludes the protein’s permanent dipole moment (rather than the induced moment) from contributing to the DEP force. Based on the magnitude of the β-dispersion exhibited by globular proteins in the frequency range 1 kHz–50 MHz, an empirically derived molecular version of CM is obtained. This factor varies greatly in magnitude from protein to protein (e.g., ~37,000 for carboxypeptidase; ~190 for phospholipase) and when incorporated into the basic expression for the DEP force brings most of the reported protein DEP above the minimum required to overcome dispersive Brownian thermal effects. We believe this empirically-derived finding validates the theories currently being advanced by Matyushov and co-workers.
New theoretical relationships are derived to link the dielectric properties of a suspension of colloidal particles to both the dielectrophoretic (DEP) and electrorotation (ROT) behaviour exhibited by a single suspended particle. It is found that the relaxation frequencies that characterize the dielectric spectrum of a colloidal suspension are close to, but different from, those that characterize the DEP and ROT responses. The extent of this difference is dependent on particle volume fraction and the intrinsic dielectric properties of both particle and suspending medium. Experimental results obtained for yeast cells in the frequency range from 1 kHz to 10 MHz provide confirmation of the theory.
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