Spectrophotometric Studies on the Disk-Sphere Transformation of Red Blood Cell by Alkali

Abstract

Although mammalian red cells, in general, have biconcave discoid forms, they change readily their shapes, however, in saline solution and especially in the “oligolytic concentration”, 0.4mol, assume promptly crenated spheres. Furchgott and Ponder reported that the biconcave discoid erythrocytes in the physiological saline soultion change the shapes easily by changing the pH of the medium and become immediately spheric in alkali.The authors investigated the disk-sphere transformation of the red cell in various concentrations of saline by adding sodium hydroxide and studies the shape change from the view-point of the light absorption of the erythrocyte suspension.The spheric transformation of the 0.032% erythrocyte suspension by the addition of alkali, especially 1/1000N to 1/12000N sodium hydroxide, was studied spectrophotometrically. The shape change of the red cell could be detected by the light absorption curve as in the case of the solution of the neutral salt.1) The red cell assumes readily the complete spheric form by the addition of sodium hydroxide in the lower concentration than the oligolytic concentration and the more readily with the decreased concentration of saline. At the same time, the light absorption curve of the erythrocyte suspension shows an decrease in short wave length.2) The decrease in the light absorption curve in short wave length, however, is small with the decreased concentration of saline even if the red cells are the spheres of same size, and at the same time the effect of the absorption bands of hemoglobin contained in the red cell is observed a little.3) When sodium hydroxide is added to the erythrocyte suspension, the red cell is transformed spheric and hemolyzed in several minutes or hours according to the concentration of sodium hydroxide. If the concentration of sodium hydroxide is high, the red cell assumes again the biconcave discoid form just before hemolysis occurs. The phenomenon is also related to the concentration of saline as a dispersion medium and the disk transformation occurs by the more diluted sodium hydroxide with the decreased concentration of saline, but occurs hardly with the increased concentration of saline. When the red cell becomes again biconcave discoid, the light absorption curve increases in short wave length and the obvious absorption bands by the effect of hemoglobin are obseved in 420, 540 and 580mμ.4) In the case that the dispersion medium consists only of saline solution, the light absorption of the erythrocyte suspension shows in the oligolytic concentration, 0.4mol, the remarkable maximum in 720mμ and minimum in 420mμ. When sodium hydroxide is added to the dispersion medium, the light absorption in the lower concentration than the oligolytic concentration shows remarkable change, which is decrease of the light absorption in 420mμ and increase in 720mμ. In this case the maximum of minimum is not obvious.5) And the change of the light absorption in the lower concentration than the oligolytic concentration is more conspicuous with the stronger bases in the same normality.