Ivanov I B, Hadjiiski A, Denkov N D, Gurkov T D, Kralchevsky P A, Koyasu S
Laboratory of Thermodynamics and Physico-chemical Hydrodynamics, Faculty of Chemistry, University of Sofia, 1126 Sofia, Bulgaria.
Biophys J. 1998 Jul;75(1):545-56. doi: 10.1016/S0006-3495(98)77544-7.
A novel method for studying the interaction of biological cells with interfaces (e.g., adsorption monolayers of antibodies) is developed. The method is called the film trapping technique because the cell is trapped within an aqueous film of equilibrium thickness smaller than the cell diameter. A liquid film of uneven thickness is formed around the trapped cell. When observed in reflected monochromatic light, this film exhibits an interference pattern of concentric bright and dark fringes. From the radii of the fringes one can restore the shape of interfaces and the cell. Furthermore, one can calculate the adhesive energy between the cell membrane and the aqueous film surface (which is covered by a layer of adsorbed proteins and/or specific ligands), as well as the disjoining pressure, representing the force of interaction per unit area of the latter film. The method is applied to two human T cell lines: Jurkat and its T cell receptor negative (TCR-) derivative. The interaction of these cells with monolayers of three different monoclonal antibodies adsorbed at a water-air interface is studied. The results show that the adhesive energy is considerable (above 0.5 mJ/m2) when the adsorption monolayer contains antibodies acting as specific ligands for the receptors expressed on the cell surface. In contrast, the adhesive energy is close to zero in the absence of such a specific ligand-receptor interaction. In principle, the method can be applied to the study of the interaction of a variety of biological cells (B cells, natural killer cells, red blood cells, etc.) with adsorption monolayers of various biologically active molecules. In particular, film trapping provides a tool for the gentle micromanipulation of cells and for monitoring of processes (say the activation of a T lymphocyte) occurring at the single-cell level.
开发了一种研究生物细胞与界面(如抗体吸附单层)相互作用的新方法。该方法被称为膜捕获技术,因为细胞被困在平衡厚度小于细胞直径的水膜内。在被困细胞周围形成厚度不均匀的液膜。当在反射单色光下观察时,该液膜呈现出同心亮条纹和暗条纹的干涉图案。从条纹半径可以恢复界面和细胞的形状。此外,可以计算细胞膜与水膜表面(覆盖有一层吸附蛋白和/或特定配体)之间的粘附能,以及代表后一种膜单位面积相互作用力的分离压力。该方法应用于两种人类T细胞系:Jurkat及其T细胞受体阴性(TCR-)衍生物。研究了这些细胞与吸附在水-空气界面的三种不同单克隆抗体单层的相互作用。结果表明,当吸附单层包含作为细胞表面表达受体的特异性配体的抗体时,粘附能相当可观(高于0.5 mJ/m²)。相反,在没有这种特异性配体-受体相互作用的情况下,粘附能接近于零。原则上,该方法可应用于研究各种生物细胞(B细胞、自然杀伤细胞、红细胞等)与各种生物活性分子吸附单层的相互作用。特别是,膜捕获为细胞的温和微操作以及监测单细胞水平发生的过程(如T淋巴细胞的激活)提供了一种工具。