Electrostatic properties of cells estimated by absorption and fluorescence spectroscopy.
A colloid titration method was used to determine the surface charge of cells of a human colon adenocarcinoma cell line WiDr; 6.2 +/- 0.8 degrees - 108 charges per cell were found. The apparent surface charge density was calculated using the cell surface area estimated by a Coulter counter. Alternatively, the lower limit of the cell surface area was estimated by visible microscopy. The same procedure was applied for human skin fibroblasts, resulting in the value 9.4 +/- 1.1 x 108 charges per cell. This is significantly higher (p < 0.05) than that of WiDr cells, presumably because of the different size of the cells. According to the estimations using the Coulter counter, the median diameter was higher in the case of skin fibroblasts. Fluorimetric titration of the fluorescent probe U-6 was used to estimate the interfacial potential of the WiDr cells. A shift of the titration curve of the U-6 probe toward higher pH values compared to that in pure buffer solutions was found in the presence of the WiDr cells. From the displacement of the midpoints of the titration curves, the interfacial potential of the WiDr cells was found to be about -35.8 mV. Incubation of the cells at two different pH values (7.4 and 6.8) did not result in any significant modification of the electrostatic properties of the cells under the experimental conditions of the present study. Electron microscopy revealed a distinct difference in the surface morphology of the WiDr cells compared to human skin fibroblasts. Numerous microvilli present on the surface of WiDr cells indicated marked uncertainties in cell surface area estimations. This gives large uncertainties in the real surface charge densities of cells.
Cunderlíková
Beata
B
Department of Biophysics, Institute for Cancer Research, Montebello, N-0310 Oslo, Norway. beata@ilc.sk
Moan
Johan
J
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology; D001703:Biophysics / Q000379:methods*; D045744:Cell Line, Tumor; D003102:Colloids / Q000737:chemistry; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008855:Microscopy, Electron, Scanning / Q000379:methods; D012867:Skin / Q000378:metabolism / Q000473:pathology; D013050:Spectrometry, Fluorescence / Q000379:methods; D055672:Static Electricity; D013499:Surface Properties; D013997:Time Factors
D003102:Colloids