Garrell R L, Crepeau R H, Edelstein S J
Proc Natl Acad Sci U S A. 1979 Mar;76(3):1140-4. doi: 10.1073/pnas.76.3.1140.
Fibers of deoxyHb S have been investigated by thin-section electron microscopy, utilizing a tannic acid embedding procedure. On the basis of numerous measurements of cross-sectional center-to-center distances for adjacent fibers in pairs or arrays, fiber diameters (mean +/- SD) of 205 +/- 5 A in embedded cells and 212 +/- 8 A in embedded hemolysates were obtained. This is an agreement with values obtained by conventional embedding procedures [Crepeau, R. H., Dykes, G., Garrell, R. L. & Edelstein, S. J. (1978) Nature (London) 274, 616--617]. The use of tannic acid has resulted in improved resolution of fiber cross sections, revealing individual strands of Hb S molecules. Because the section thickness corresponds to approximately one-fifth of the fiber helical repeat distance, the strands in projection superimpose to form characteristic image patterns. Additional superposition patterns arise in sections taken at small deviations from perpendicularity to the longitudinal fiber axis. These patterns are consistent with the 14-strand structure for hemoglobin S fibers [Dykes, G., Crepeau, R. H. & Edelstein, S. J. (1978) Nature (London) 272, 506--510], as indicated by computer models of cross-sectional patterns for various thicknesses and angular deviations of sections.
利用单宁酸包埋法,通过超薄切片电子显微镜对脱氧血红蛋白S纤维进行了研究。基于对成对或成排相邻纤维的横截面中心到中心距离的大量测量,得出包埋细胞中纤维直径(平均值±标准差)为205±5埃,包埋溶血产物中为212±8埃。这与通过传统包埋法获得的值一致[Crepeau, R. H., Dykes, G., Garrell, R. L. & Edelstein, S. J. (1978) Nature (London) 274, 616 - 617]。单宁酸的使用提高了纤维横截面的分辨率,揭示了血红蛋白S分子的单链。由于切片厚度约为纤维螺旋重复距离的五分之一,投影中的链相互叠加形成特征图像模式。在与纤维纵轴垂直方向有小偏差的切片中会出现额外的叠加模式。这些模式与血红蛋白S纤维的14链结构一致[Dykes, G., Crepeau, R. H. & Edelstein, S. J. (1978) Nature (London) 272, 506 - 510],各种厚度和角度偏差切片的横截面模式计算机模型表明了这一点。