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Analysis of erythrocyte flexibility by means of laser diffraction: effects of mechanical stress, photosensitization, and ozone

A newly developed system to determine red blood cell (RBC) flexibility is presented. It combines ektacytometry (laser diffraction) with image analysis. Laser light is passed through erythrocyte (RBC) suspensions which are sheared in a Searle like viscosimeter. The laser diffraction patterns are photographed by a linear CCD camera, and further analyzed by a computer. Flexibility is characterized by the quotient of minor and major axes of isointensity lines of the elliptically transformed diffraction patterns. The variation coefficient of the measurement is less than 1%. Exposing RBC to mechanical stress within the viscosimeter (shear rates from 130 to 2620/s, viscosity of the suspending medium 22 - 24 cpoise) is followed by a decrease of RBC flexibility. This effect is only detectable if elongation is measured at low shear rates (e.g., 260/s) and a threshold of stress (1100/s) is exceeded. It is not reversible and not accompanied with detectable hemolysis. Exposing whole blood to ozone (2 - 77 (mu) g/ml) does not alter RBC flexibility, but induces significant hemolysis at ozone > 7 (mu) g/ml. Thus the action of ozone can be regarded as an all or nothing effect either leaving the cell unaffected (flexibility) or destroying it (hemolysis).