Flickering analysis of erythrocyte mechanical properties: dependence on oxygenation level, cell shape, and hydration level.

Erythrocytes (red blood cells) play an essential role in the respiratory functions of vertebrates, carrying oxygen from lungs to tissues and CO(2) from tissues to lungs. They are mechanically very soft, enabling circulation through small capillaries. The small thermally induced displacements of the membrane provide an important tool in the investigation of the mechanics of the cell membrane. However, despite numerous studies, uncertainties in the interpretation of the data, and in the values derived for the main parameters of cell mechanics, have rendered past conclusions from the fluctuation approach somewhat controversial. Here we revisit the experimental method and theoretical analysis of fluctuations, to adapt them to the case of cell contour fluctuations, which are readily observable experimentally. This enables direct measurements of membrane tension, of bending modulus, and of the viscosity of the cell cytoplasm. Of the various factors that influence the mechanical properties of the cell, we focus here on: 1), the level of oxygenation, as monitored by Raman spectrometry; 2), cell shape; and 3), the concentration of hemoglobin. The results show that, contrary to previous reports, there is no significant difference in cell tension and bending modulus between oxygenated and deoxygenated states, in line with the softness requirement for optimal circulatory flow in both states. On the other hand, tension and bending moduli of discocyte- and spherocyte-shaped cells differ markedly, in both the oxygenated and deoxygenated states. The tension in spherocytes is much higher, consistent with recent theoretical models that describe the transitions between red blood cell shapes as a function of membrane tension. Cell cytoplasmic viscosity is strongly influenced by the hydration state. The implications of these results to circulatory flow dynamics in physiological and pathological conditions are discussed.