Tishler R B, Carlson F D
Thomas C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland 21218.
Biophys J. 1993 Dec;65(6):2586-600. doi: 10.1016/S0006-3495(93)81302-X.
A quasi-elastic light-scattering (QELS) microscope spectrometer was used to study the dynamic properties of the membrane/cytoskeleton of individual human red blood cells (RBCs). QELS is a spectroscopic technique that measures intensity fluctuations of laser light scattered from a sample. The intensity fluctuations were analyzed using power spectra and the intensity autocorrelation function, g(2)(tau), which was approximated with a single exponential. The value of the correlation time, Tcorr, was used for comparing results. Motion of the RBC membrane/cytoskeleton was previously identified as the source of the QELS signal from the RBC (R. B. Tishler and F. D. Carlson, 1987. Biophys. J. 51:993-997), and additional data supporting that conclusion are presented. Similar results were obtained from anucleate mammalian RBCs that have structures similar to that of the human RBC, but not for morphologically distinct, nucleated RBCs. The effect of altering the physical properties of the cytoplasm and the membrane/cytoskeleton was also studied. Osmotically increasing the cytoplasmic viscosity led to significant increases in Tcorr. Increasing the membrane cholesterol content and increasing the intracellular calcium content both led to decreased deformability of the human RBC. In both cases, the modified cells with decreased deformability showed an increase in Tcorr, demonstrating that QELS could measure biochemically induced changes of the membrane/cytoskeleton. Physiological changes were measured in studies of age-separated RBC populations which showed that Tcorr was increased in the older, less deformable cells.
使用准弹性光散射(QELS)显微镜光谱仪研究单个人类红细胞(RBC)膜/细胞骨架的动态特性。QELS是一种光谱技术,用于测量从样品散射的激光强度波动。使用功率谱和强度自相关函数g(2)(τ)对强度波动进行分析,g(2)(τ)用单指数近似。相关时间Tcorr的值用于比较结果。红细胞膜/细胞骨架的运动先前被确定为来自红细胞的QELS信号的来源(R.B.Tishler和F.D.Carlson,1987年。生物物理学杂志。51:993 - 997),并给出了支持该结论的其他数据。从具有与人类红细胞相似结构的无核哺乳动物红细胞中获得了类似结果,但形态不同的有核红细胞则不然。还研究了改变细胞质和膜/细胞骨架物理性质的影响。渗透性增加细胞质粘度导致Tcorr显著增加。增加膜胆固醇含量和增加细胞内钙含量均导致人类红细胞的变形性降低。在这两种情况下,变形性降低的修饰细胞显示Tcorr增加,表明QELS可以测量膜/细胞骨架的生化诱导变化。在对按年龄分离的红细胞群体的研究中测量了生理变化,结果表明,在较老、变形性较小的细胞中Tcorr增加。