Dielectric properties of human leukocyte subpopulations determined by electrorotation as a cell separation criterion.
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 1 kHz to 120 MHz. 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/m2 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.
Yang
J
J
Department of Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA.
Huang
Y
Y
Wang
X
X
Wang
X B
XB
Becker
F F
FF
Gascoyne
P R
PR
D055592:Biophysical Phenomena; D001703:Biophysics; D002462:Cell Membrane / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods*; D004563:Electrochemistry; D005434:Flow Cytometry; D006801:Humans; D007962:Leukocytes / Q000145:classification* / Q000166:cytology* / Q000276:immunology; D008855:Microscopy, Electron, Scanning; D012399:Rotation