Makovitzky J
Histochemistry. 1983;79(2):269-79. doi: 10.1007/BF00489789.
The latent birefringence of lymphocyte membranes of various species may readily be studied and analysed by various topo-optical reactions. The membranes of glutaraldehyde-fixed and PBS-washed lymphocytes show continuous birefringence with thiazine- and quinoline dyes. According to polarization optical analysis thiazine dye-stained cells are radially positive, whereas quinoline dye-stained cells are radially negative spherites, i.e. thiazine dye molecules are in a perpendicular, quinoline dye molecules in a parallel orientation relative to the membrane surface. These findings suggest that in lymphocyte membranes glycoproteins are primarily responsible for the topo-optical reactions. The actual conformational state of the glycoprotein components is a decisive factor not only in dye binding but also in the orientation of dye molecules. Heparin treatment directs attention to an important interaction between heparin and membrane glycoproteins. With the aid of the critical electrolyte concentration (CEC) technique we were able to demonstrate an ultrastructural differences between human erythrocyte and human lymphocyte membranes. After this procedure the birefringence of erythrocyte membranes was lost, whereas that of lymphocyte membranes did not change. There were no differences between the topo-optical reactions of T and B lymphocytes.
通过各种拓扑光学反应,可以很容易地研究和分析不同物种淋巴细胞膜的潜在双折射。经戊二醛固定并用磷酸盐缓冲盐水(PBS)洗涤的淋巴细胞膜,用噻嗪染料和喹啉染料染色后呈现连续双折射。根据偏振光学分析,噻嗪染料染色的细胞呈径向阳性,而喹啉染料染色的细胞是径向阴性球体,即噻嗪染料分子相对于膜表面呈垂直取向,喹啉染料分子呈平行取向。这些发现表明,淋巴细胞膜中的糖蛋白是拓扑光学反应的主要原因。糖蛋白成分的实际构象状态不仅是染料结合的决定性因素,也是染料分子取向的决定性因素。肝素处理使人们关注到肝素与膜糖蛋白之间的重要相互作用。借助临界电解质浓度(CEC)技术,我们能够证明人类红细胞膜和人类淋巴细胞膜之间的超微结构差异。经过该程序后,红细胞膜的双折射消失,而淋巴细胞膜的双折射没有变化。T淋巴细胞和B淋巴细胞的拓扑光学反应没有差异。