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We present a new method to measure the shear elastic moduli and viscosities of erythrocyte membranes which is based on the fixation and transient deformation of cells in a high-frequency electric field. A frequency domain of constant force (arising by Maxwell Wagner polarization) is selected to minimize dissipative effects. The electric force is thus calculated by electrostatic principles by considering the cell as a conducting body in a dielectric fluid and neglecting membrane polarization effects. The elongation A of the cells perpendicular to their rotational axis exhibits a linear regime (A proportional to Maxwell tension or to square of the electric field E2) at small, and a nonlinear regime (A proportional to square root of Maxwell tension or to the electric field E) at large extensions with a cross-over at A approximately 0.5 micron. The nonlinearity leads to amplitude-dependent response times and to differences of the viscoelastic response and relaxation functions. The cells exhibit pronounced yet completely reversible tip formations at large extensions. Absolute values of the shear elastic modulus, mu, and membrane viscosity, eta, are determined by assuming that field-induced stretching of the biconcave cell may be approximately described in terms of a sphere to ellipsoid deformation. The (nonlinear) elongation-vs.-force relationship calculated by the elastic theory of shells agress well with the experimentally observed curves and the values of mu = 6.1 x 10(-6) N/m and eta = 3.4 x 10(-7) Ns/m are in good agreement with the micropipette results of Evans and co-workers. The effect of physical, biochemical, and disease-induced structural changes on the viscoelastic parameters is studied. The variability of mu and eta of a cell population of a healthy donor is +/- 45%, which is mainly due to differences in the cell age. The average mu value of cells of different healthy donors scatters by +/- 18%. Osmotic deflation of the cells leads to a fivefold increase of mu and 10-fold increase of eta at 500 mosm. The shear modulus mu increases with temperature showing that the cytoskeleton does not behave as a network of entropy elastic springs. Elliptic cells of patients suffering from elliptocytosis of the Leach phenotype exhibit a threefold larger value of mu than normal discocytes of control donors. Cross-linking of the spectrin by the divalent S-H agents diamide (1 mM, 15 min incubation) leads to an eightfold increase of mu whereas eta is essentially constant. The effect of diamide is reversed after treatment with S-S bond splitting agents.

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The high deformability of erythrocytes which is essential for their transport through the capillaries depends critically on their discoid shape and on the elasticity of the plasma membrane, which may be determined by interactions of the cytoskeleton, the lipid/protein leaflet and the glycocalyx. Although techniques exist for measurement of the static elastic properties of erythrocytes, the cells are continuously deformed in vivo, the stress varying within periods of a few seconds. Thus dynamic elastic behaviour is essential for their physiological function. We present here a novel means of measuring the dynamic elastic constants of the red cell based on the transient deformation of individual cells in an inhomogeneous high-frequency (HF) electric field. By microscopy it is possible to record cellular elongations as small as 200 nm occurring within time scales of 1 ms. A main advantage is that the cellular response is linear and thus can be more readily interpreted theoretically. We have observed a creep function consisting of two exponentials with response times of 0.1 s and 1 s, which can be described in terms of a simple viscoelastic model. A remarkable temperature dependence of the membrane elasticity between 25 degrees C and 15 degrees C is observed for freshly drawn cells but not for trypsinized ones.

2014 Nous mesurons l'élasticité de flexion de vésicules flasques par l'analyse de Fourier des ondulations thermiques de surface, jusqu'au 8e ordre. Les résultats sont expliqués par la théorie récente de Schneider et al. La constante élastique de flexion pour une bicouche fluide de lécithine (Kc = 4 x 10-13 erg) est en bon accord avec celle des membranes d'érythrocytes, mais est presque d'un ordre de grandeur plus faible que celle donnée pour les mêmes lipides dans la littérature. Ce désaccord est attribué au fait que la valeur de Kc obtenue en ne considérant que le mode fondamental de déformation est systématiquement trop élevée. Abstract. 2014 We measure the bending elasticity of flaccid vesicles by Fourier analysis of the thermally excited surface undulations up to the 8th order. The results are explained in terms of the recent theory of Schneider et al. The value of the bending elastic constant for fluid lecithin bilayers (Kc = 4 x 10-13 erg) agrees well with that of erythrocyte membranes but is nearly an order of magnitude smaller than that reported for the same lipid in the literature. This discrepancy is attributed to the fact that the Kc value obtained by considering the lowest order mode of deformation alone is systematically too high.