| Electrorotation Studies of Baby Hamster Kidney Fibroblasts Infected with Herpes Simplex Virus Type 1 Biophysical Journal, Volume 76, Issue 5, 1 May 1999, Pages 2833-2842 Steffen Archer, Hywel Morgan and Frazer J. Rixon Abstract The dielectric properties of baby hamster kidney fibroblast (BHK(C-13)) cells have been measured using electrorotation before and after infection with herpes simplex virus type 1 (HSV-1). The dielectric properties and morphology of the cells were investigated as a function of time after infection. The mean specific capacitance of the uninfected cells was 2.0F/cm, reducing to a value of 1.5F/cm at 12h after infection. This change was interpreted as arising from changes in the cell membrane morphology coupled with alterations in the composition of the cell membrane as infection progressed. The measured changes in the cell capacitance were correlated with alterations in cellular morphology determined from scanning electron microscope (SEM) images. Between 9 and 12h after infection the internal permittivity of the cell exhibited a rapid change, reducing in value from 75 to 58, which can be correlated with the generation of large numbers of Golgi-derived membrane vesicles and enveloped viral capsids. The data are discussed in relation to the known life cycle of HSV-1 and indicate that electrorotation can be used to observe dynamic changes in both the dielectric and morphological properties of virus-infected cells. Calculations of the dielectrophoretic spectrum of uninfected and infected cells have been performed, and the results show that cells in the two states could be separated using appropriate frequencies and electrode arrays. Abstract | Full Text | PDF (220 kb) |
| Dielectric Properties of Human Leukocyte Subpopulations Determined by Electrorotation as a Cell Separation Criterion Biophysical Journal, Volume 76, Issue 6, 1 June 1999, Pages 3307-3314 Jun Yang, Ying Huang, Xujing Wang, Xiao-Bo Wang, Frederick F. Becker and Peter R.C. Gascoyne Abstract The separation and purification of human blood cell subpopulations is an essential step in many biomedical applications. New dielectrophoretic fractionation methods have great potential for cell discrimination and manipulation, both for microscale diagnostic applications and for much larger scale clinical problems. To discover whether human leukocyte subpopulations might be separable by such methods, the dielectric characteristics of the four main leukocyte subpopulations, namely, B- and T-lymphocytes, monocytes, and granulocytes, were measured by electrorotation over the frequency range 1kHz to 120MHz. The subpopulations were derived from human peripheral blood by magnetically activated cell sorting (MACS) and sheep erythrocyte rosetting methods, and the quality of cell fractions was checked by flow cytometry. Mean specific membrane capacitance values were calculated from the electrorotation data as 10.5 (± 3.1), 12.6 (± 3.5), 15.3 (± 4.3), and 11.0 (± 3.2) mF/m for T- and B-lymphocytes, monocytes, and granulocytes, respectively, according to a single-shell dielectric model. In agreement with earlier findings, these values correlated with the richness of the surface morphologies of the different cell types, as revealed by scanning electron microscopy (SEM). The data reveal that dielectrophoretic cell sorters should have the ability to discriminate between, and to separate, leukocyte subpopulations under appropriate conditions. Abstract | Full Text | PDF (575 kb) |
| A Unified Resistor-Capacitor Model for Impedance, Dielectrophoresis, Electrorotation, and Induced Transmembrane Potential Biophysical Journal, Volume 75, Issue 2, 1 August 1998, Pages 1107-1116 Jan Gimsa and Derk Wachner Abstract Dielectric properties of suspended cells are explored by analysis of the frequency-dependent response to electric fields. Impedance (IMP) registers the electric response, and kinetic phenomena like orientation, translation, deformation, or rotation can also be analyzed. All responses can generally be described by a unified theory. This is demonstrated by an RC model for the structural polarizations of biological cells, allowing intuitive comparison of the IMP, dielectrophoresis (DP), and electrorotation (ER) methods. For derivations, cells of prismatic geometry embedded in elementary cubes formed by the external solution were assumed. All geometrical constituents of the model were described by parallel circuits of a capacitor and a resistor. The IMP of the suspension is given by a meshwork of elementary cubes. Each elementary cube was modeled by two branches describing the current flow through and around the cell. To model DP and ER, the external branch was subdivided to obtain a reference potential. Real and imaginary parts of the potential difference of the cell surface and the reference reflect the frequency behavior of DP and ER. The scheme resembles an unbalanced Wheatstone bridge, in which IMP measures the current-voltage behavior of the feed signal and DP and ER are the measuring signal. Model predictions were consistent with IMP, DP, and ER experiments on human red cells, as well as with the frequency dependence of field-induced hemolysis. The influential radius concept is proposed, which allows easy derivation of simplified equations for the characteristic properties of a spherical single-shell model on the basis of the RC model. Abstract | Full Text | PDF (203 kb) |
Copyright © 1996 The Biophysical Society. All rights reserved.
Biophysical Journal, Volume 71, Issue 1, 495-506, 1 July 1996
doi:10.1016/S0006-3495(96)79251-2
Research Article
J. Gimsa, T. Müller, T. Schnelle and G. Fuhr
Institute of Biology, Humboldt-University, Berlin, Germany. jan=gimsa@rz.hu-berlin.de
Usually dielectrophoretic and electrorotation measurements are carried out at low ionic strength to reduce electrolysis and heat production. Such problems are minimized in microelectrode chambers. In a planar ultramicroelectrode chamber fabricated by semiconductor technology, we were able to measure the dielectric properties of human red blood cells in the frequency range from 2 kHz to 200 MHz up to physiological ion concentrations. At low ionic strength, red cells exhibit a typical electrorotation spectrum with an antifield rotation peak at low frequencies and a cofield rotation peak at higher ones. With increasing medium conductivity, both electrorotational peaks shift toward higher frequencies. The cofield peak becomes antifield for conductivities higher than 0.5 S/m. Because the polarizability of the external medium at these ionic strengths becomes similar to that of the cytoplasm, properties can be measured more sensitively. The critical dielectrophoretic frequencies were also determined. From our measurements, in the wide conductivity range from 2 mS/m to 1.5 S/m we propose a single-shell erythrocyte model. This pictures the cell as an oblate spheroid with a long semiaxis of 3.3 microns and an axial ratio of 1:2. Its membrane exhibits a capacitance of 0.997 x 10(-2) F/m2 and a specific conductance of 480 S/m2. The cytoplasmic parameters, a conductivity of 0.4 S/m at a dielectric constant of 212, disperse around 15 MHz to become 0.535 S/m and 50, respectively. We attribute this cytoplasmic dispersion to hemoglobin and cytoplasmic ion properties. In electrorotation measurements at about 60 MHz, an unexpectedly low rotation speed was observed. Around 180 MHz, the speed increased dramatically. By analysis of the electric chamber circuit properties, we were able to show that these effects are not due to cell polarization but are instead caused by a dramatic increase in the chamber field strength around 180 MHz. Although the chamber exhibits a resonance around 180 MHz, the harmonic content of the square-topped driving signals generates distortions of electrorotational spectra at far lower frequencies. Possible technological applications of chamber resonances are mentioned.